Consumption of sugar-sweetened beverages (SSBs) or sugary drinks may reduce or even eliminate the household income allocation for other essential commodities. Reducing expenditure for consumption of other household commodities is known as the crowding-out effect of SSB. We aimed to determine the crowding-out effect of SSB expenditure on other household commodities. In addition, we also identified the factors influencing the household's decision to purchase of SSBs.We used the logistic regression  and the Seemingly Unrelated Regression (SUR) models. In order to find the probability of a given change in the socio-demographic variables, we also estimated the average marginal effects from the logistic regression. In addition, we regressed the SUR model by gender differences. We used Household Income and Expenditure Survey (HIES) 2016 data to estimate our chosen econometric models. HIES is nationally representative data on the household level across the country and is conducted using a multistage random sampling method by covering 46,075 households.The findings from the logit model describe that the greater proportion of male members, larger household size, household heads with higher education, profession, having a refrigerator, members living outside of the house, and households with higher income positively affect the decision of purchasing SSB. However, the determinants vary with the various types of SSB. The unadjusted crowding out effect shows that expenditure on SSB or sugar-added drinks crowds out the household expenditure on food, clothing, housing, and energy items. On the other hand, the adjusted crowding out effect crowds out the spending on housing, education, transportation, and social and state responsibilities.Although the household expenditure on beverages and sugar-added drinks is still moderate (around 2% of monthly household expenditure), the increased spending on beverages and sugar-added drinks is a concern due to the displacement of household expenditure for basic commodities such as food, clothing, housing, education, and energy. Therefore, evidence-based policies to regulate the sale and consumption of SSB are required for a healthy nation. The consumption pattern of sugar-sweetened beverages (SSBs) or sugary drinks is growing worldwide. Liquids sweetened with various forms of added sugar are commonly known as SSB, such as regular soda, fruit drinks, sports and energy drinks, sweetened waters, pop, cola, tonic fruit punch, lemonade, sweetened powdered drinks, and coffee and tea beverages with added sugars . The SSBGrowing consumption of SSBs not only impacts human health but also affects the household resource allocation for basic goods and services. Consumption of SSBs includes both direct (out-of-pocket expenditure) and indirect costs  to the consumer and the society in the process of treating obesity, metabolic syndromes , and kidney disease . ConsumpThe rate of SSB consumption is alarming in Bangladesh, especially among the youth and children. Evidence suggests that 48% of the school children consumed soft drinks on a daily basis , while mWe have wielded Household Income and Expenditure Survey (HIES) 2016 to explore the determinants and crowding-out effect of Sugar-Sweetened Beverages (SSB) or sugary drinks in Bangladesh. HIES data is nationally representative and is collected by the Bangladesh Bureau of Statistics (BBS) at the household level at five-year intervals since 1973–74 . We haveSugar-sweetened beverages (SSBs) generally include soft drinks, sugar-added fruit beverages, carbonated drinks, regular sugary soda, and sweetened waters . Tea andWe have used a logistic regression model to delineate the determinants of consuming sugar-sweetened beverages and other drinks. On the other hand, we have exercised the Seemingly Unrelated Regression (SUR) model to depict the crowding-out effect of sugar-sweetened beverages following a similar kind of study . We firsd indicates the status of a household consuming a particular kind of drink  is used more frequently to avoid the above problem .3\documeThus, AME can be derived by taking the arithmetic mean of each unit of marginal effects.The crowding-out effect of SSB has been defined as the households' reduced consumption of basic necessities due to the consumption of beverages and other sugar-added drinks. On the other hand, crowding out of certain goods and commodities may be derived as the difference of average expenditure shares of different consumption categories between SSBs consumers and non-consumer , and/or The existing literature estimates the crowding out effect in two ways: without controlling (unadjusted) the household socioeconomic characteristics and by controlling (adjusted) them. In the former model, we have compared the mean expenditure share of the households that consume drinks and the households that do not consume any drinks without controlling the socio-demographic variables of the individuals and households. We have also tested the mean differences using the t-test. The negative statistically significant differences are termed as the unadjusted crowding out effect of drink consumption, whereas the positive one increases the expenditure. In contrast, the second model has been estimated based on the household utility maximization problem for the purchasing decision of households between sugar-added drinks and other basic necessities . Therefod, and 0 otherwise. There are 8 administrative division in Bangladesh, and we assume prices are fixed within the division. The Quadratic Almost Ideal Demand System (QAIDS) framework  is widelOn the other hand, to estimate the crowding out effect of only soft drinks, only other sugar-added drinks, and only both drinks, we have used the following equation:Equation  and 4.24.2 are tSimilar studies of crowding out , 26, 27 Since the household decision to purchase goods and services is made simultaneously, the expenditure of one item is more likely correlated with other categories. Therefore, we may also assume that the dependent variables will be correlated with the error term of the other equations (contemporaneous correlation) . Each ofTable It is also evident that urban people are more inclined to consume beverages and other drinks than rural people. In addition, a greater proportion of the households in the top quantile consume SSB compared to the proportion of the households in the bottom quantile, regardless of the households' residential location.Table Similarly, a greater proportion of households consuming SSB are male-headed (95%), compared to the households (81%) with no expenditure on any SSB. However, the proportion of adult members in the households is lower in the households with expenditure on SSB, though the proportion of adult males among adult members and the proportion of earners in the households are approximately identical Table .Regarding the health consequences, a larger proportion of the households that have spent on SSB (51%) had at least one member who suffered from any chronic disease during the last twelve months than the household with no expenditure on any drinks (45%). Since households that consume SSB may need refrigerators to store them, they may possess refrigerators more likely than households with no drink expenditure. Table We found that the higher proportion of adult members, having sick members, larger household size, urban location of the households, household heads' education, non-agriculture profession of the household head, having a refrigerator, members living outside of the house, and households income positively affect the decision of purchasing SSB Table . If the Although the proportion of adult members and male-headed households increases the likelihood of the decision to purchase any SSBs or sugary drinks, they decrease the probability for only soft drinks beverages (column 3). However, though the sign and significance for both drinks (column 4) are similar to any drinks (column 1), some deviations are found in only other sugar-added drinks (column 3). For instance, the sign of the refrigerator coefficients is found to be negative. It might happen because the drinks items in other sugar-added drinks may not require the necessity of refrigerators for storage purposes.Table Table According to column 3 of Table Some differences are found in the magnitudes, signs, and significance levels of the coefficients for the SSB expenditure by type. For instance, the coefficients of the housing category for the household with expenditure only soft drinks and both drinks are found positive and negative, respectively, without being statistically significant. The magnitude of food items for the household with having expenditure only on both types of drinks is almost double (-3.24) than the other categories. It implies that expenditure on both kinds of drinks crowds out the larger portion of the food budget for the households than other categories.Since the estimated results of Table The estimated adjusted crowding out results (column 2) due to the expenditure for SSB by types in household different consumption categories are almost similar to the crowding out effect due to the expenditure on any SSB consumption. However, there are some variations in magnitude, signs, and level of significance among the different types of models, like the unadjusted crowding-out effect. The most apparent difference is found in the magnitude of the food items (8.63) of the household's expenditure on only beverages and other sugar-added drinks.Table In both Tables Households may have zero consumption of SSBs due to the lack of income. It implies that poor households may have corner solutions for SSBs consumption. Existing literature also shows that poorer households have significantly lower SSBs consumption than richer households , 29. TheWe estimated the crowding-out effect without controlling the covariates (unadjusted crowding out) and by controlling the covariates (adjusted crowding out). The unadjusted crowding-out effects reveal that expenditure on SSB crowds out the expenditure on food, clothing, housing, and social responsibility. In contrast, the adjusted crowding-out effects present that the expenditure on SSB crowds out the expenditure on housing, education, transportation, and social responsibility. In both cases, it is evident that SSB consumption displaces the household expenditure for basic necessities such as food, clothing, housing, transportation, and education. It implies that the consumption of SSB affects human capital development, and therefore, it hinders the well-being of the people. In a similar study on tobacco, Hussain et al. 2018) [ [18] alsThe findings present that households with SSB consumption are more likely to have sick members who suffered from any chronic disease during the last twelve months. On the other hand, logistic regression results show that households with sick members are positively associated with consuming SSB or other sugar-added drinks. Unadjusted crowding out effects of SSB and sugar-added drinks depicts that the households that have expenditure on only soft drinks have greater expenditure on medicine, implying that SSB consumption poses a health burden. However, the coefficients for all specifications for adjusted crowding out are found positive, though they are not statistically significant. It insinuates that consumption of SSB increases the risk of being affected with chronic diseases, and therefore, it increases the expenses for medicine. It is evident that the consumption of SSB has also caused budget displacement in the household income portfolio.Our econometric results also show that male-headed households are more likely to allocate their income to beverages and other sugar-added drink commodities. Therefore, crowding out for various food and non-food items is more prevalent for household consumption items in male-headed households. It seems rational because male members of the households remain mostly outside and therefore are more frequently used to beverages and sugar-added drinks than the female members.The study has a number of limitations. As the latest HIES 2021 data is not publicly available yet, the study team analysed the HIES 2016 data. There might be changes in the SSB consumption pattern and related variables since 2016. Moreover, the available SSBs in Bangladesh are diverse in nature, and they are not well defined in Bangladesh for policy and practice. The HIES 2016 data also does not define 'SSB' explicitly. For instance, sherbats (like fruit juices) are added to the category of soft drinks (code 151) in the current HIES 2016. In addition, bottled waters are added with the soft drinks (code 194) in the dining out section. Therefore, it is not possible to disaggregate the sherbats from the soft drinks, or to determine whether the SSB or bottled water was consumed. In the absence of an operational definition of SSB and limitations of the available data, we have categorized the SSB in line with the questions asked in the HIES 2016 survey and their classification of SSBs. Despite the limitations, this study, for the first time, generates evidence of the crowding-out effect of sugar-sweetened beverages (SSBs) on household expenditure patterns in Bangladesh. We recommend that for future rounds of HIES, the SSB needs to be defined properly and collect the disaggregated response accordingly. This is also important for designing taxation on diverse types of SSB products and designing an intervention for all kinds of SSBs.Since the consumption of SSB negatively affects the basic household consumption and well-being of the people, evidence-based policies are required in order to curb the use and overuse of SSB. Future research is needed to design and implement innovative interventions to curb the consumption of SSB.The increased consumption of sugar-sweetened beverages (SSBs) not only impacts human health but also crowds out the household income allocation to the basic necessities. Wielding nationally representative Household Income and Expenditure Survey (HIES) 2016 data, we found that male members, larger household sizes, household heads with higher education, profession, having a refrigerator, members living outside of the house, and households with higher income positively affect the decision of purchasing SSB. In addition, the consumption of SSB crowds out the household spending for basic necessities like food, clothing, housing, education, transportation, and social responsibilities. Based on our findings, we recommend evidence-based policies to regulate the sale and consumption of SSB for a healthy and productive nation.


The complex interaction between microorganisms, the host’s immune response, and the release of pro- and anti-inflammatory factors influence the evolution of sepsis. According to the current definition, sepsis should be considered a severe, potentially fatal organ dysfunction caused by an inadequate or dysregulated host response to infection . OverallFrom a pathophysiological point of view, the interaction between some structural components (such as lipopolysaccharide for Gram-negative bacteria or lipotecoic acid for Gram-positive bacteria) and the production of bacterial exotoxins can cause a systemic activation of the immunological compound, also found through the expression of superantigens. Specifically, TLRs (Toll-like receptors) expressed on monocytes and macrophages recognize PAMPs (extracellular pathogen-associated molecular patterns) and DAMPs (damage-associated molecular patterns) released from damaged endogenous cells. As a result of this mechanism, massive amounts of pro-inflammatory cytokines are released, resulting in increased nitric oxide (NO) production, capillary permeability and vasodilation, and thus hypotension. Impaired coagulation, if massive, can lead to the development of disseminated intravascular coagulation with the excessive consumption of coagulation factors and platelets. Vasodilation, the extravasation of fluids, and disseminated thromboses have, as an evident consequence, hypoperfusions of organs and tissues, resulting in a possible development of multi-organ failure. Many studies highlight the need for a clearer understanding of the pathophysiological mechanisms underlying sepsis in order to achieve an early and timely diagnosis and to set up strategic therapies that are increasingly focused on precision medicine. The recent Special Issue “The Interplay between Host Defense, Infection and Pathogen Virulence in Sepsis and Septic Shock” highlighted important aspects of the diagnoses and management of septic patients. As reported above, the dysregulated host response to infection, leading to sepsis and septic shock, is a life-threatening event that is associated with high mortality rate, despite advances in organ support and antimicrobial therapy. Despite the implementation of international guidelines supporting early goal directed therapy, recent randomized trials have demonstrated that these interventions do not improve the survival of septic patients.This evidence warrants an urgent clarification of the molecular mechanisms underlying clinical response in patients with sepsis or septic shock. The key to improving these processes lies in acquiring in-depth knowledge of the intricate interplay between host defense, infection, and pathogen virulence, as well as the timing and type of interventions that are most effective according to the personal characteristics of individual patients. Of importance, the pharmacokinetic (PK) and pharmacodynamic (PD) properties of antibiotics should be considered because of changes in clearance and volume of distribution that are frequently observed in critically ill patients, with the potential to influence the concentration of the drug at the sites of infection.(1)the long-term effects of chronic neonatal inflammation in the course of neonatal sepsis and thus the need for an early diagnosis;(2)the attempt to identify new diagnostic strategies using more sensitive and specific biomarkers aimed at achieving increasingly rapid health interventions and therapeutic improvements;(3)the limitations of current diagnostic techniques, especially during polymicrobial sepsis;(4)the key roles of the pathophysiological mechanisms of sepsis to identify new diagnostic and therapeutic strategies, especially for multidrug resistant (MDR) pathogens.Based on the aforementioned knowledge of the mechanisms related to the progression from sepsis to septic shock and the adequate management of patients, including choice and dosages of antimicrobials, the results are crucial to improve outcome of septic patients. Specifically, the manuscripts published in this Special Issue ,5,6,7,8 It should be considered that sepsis causes high rates of morbidity and mortality also in the neonatal intensive care unit (NICU). The review conducted by Maddaloni et al.  focused Staphylococcus epidermidis, a ubiquitous skin commensal, is reported to be the main etiology in late-onset neonatal sepsis (3–28 days after birth). It is unclear what long-term chronic neonatal inflammation might entail, but it is probable that the damage could affect cognitive pathways, impairing neuronal development. Thus, laying the groundwork for the early diagnosis of neonatal sepsis in preterm infants remains a challenge and a goal to be achieved. This is because, even today, diagnosis is based on microbial blood cultures that are burdened by numerous limitations. Medical advances and molecular and sequencing techniques are needed for the development of alternative preventive, diagnostic, and therapeutic tools to combat neonatal infections. Joubert et al.  focused As a matter of fact, the management of critically ill patients admitted to an intensive care unit (ICU) remains a challenge for physicians. The majority of patients undergo continuous renal replacement therapy (CRRT) and/or extracorporeal membrane oxygenation (ECMO) that can directly affect the response to sepsis and the efficacy of antibiotic therapies. Köhler et al.  remarkedThe role of precision medicine was the topic addressed by Webber et al. . The autAnother important topic was focused on by Doualeh et al. , who shoAcinetobacter baumannii (CRAB), which is responsible for lethal sepsis and high mortality rates in hospital settings. The authors focused on attempting to develop new therapeutic options, such as the development of a novel peptide (R-Pro9-3D) with cytotoxic, antibacterial, and immunosuppressive effects, which act directly on bacterial LPS. Specifically, the study found that R-Pro9-3D had an inhibitory effect on NO production, which was responsible for acute and chronic hyperinflammation, and thus its inhibition appeared to counteract the release of pro-inflammatory factors and the organ damage that inevitably resulted.The review by Krishnan et al.  focused Finally, the review carried out by Lazzaro et al.  shed sigAcinetobacter baumannii (MDR-AB) and carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) has been observed. It is important, therefore, to develop new diagnostic techniques to obtain a fast microbiology with early microbiological reports. These techniques can include nucleic acid amplification technologies (NAATs) that amplify nucleic acid sequences to a detectable level and identify the infecting agent or the status of the immune response. The detection of bacterial DNA fragments by a real-time polymerase chain reaction (RT-PCR) in blood samples and the detection of 16S rRNA fragments of Gram-positive and Gram-negative bacteria or 18S rRNA fragments of Candida spp. seem to be very promising for shortening pathogen identification, as they have shown high degrees of specificity and sensitivity, thus reducing mortality and the length of hospitalization and ICU stay of patients. However, these techniques are not sufficient to differentiate sepsis from other inflammatory processes, and there are no biomarkers able to identify only septic patients. In recent years, an increased frequency of Gram-negative MDR pathogens such as MDR Of importance, as also reported by Lazzaro et al., the two cornerstones of sepsis treatment are empiric antibiotic therapy and infection control. Numerous studies have shown that appropriate initial antibiotic therapy reduces mortality rates more than any other medical procedure, and many studies suggest that mortality rates progressively increase, considering a diagnostic window from 1 to 24 h from the symptom-based recognition of sepsis to the start of antimicrobial treatment . Until tIn conclusion, crucial aspects of sepsis were highlighted in this Special Issue, including the mechanisms underlying sepsis; the risk factors for the development of sepsis and septic shock; the advances in diagnoses, managements, and therapies of these severe infections; and the role of adjunctive therapies for the treatment of septic patients. A timely empirical antimicrobial treatment and the importance of definitive anti-infective therapy with in vitro activity against the microbial isolates are crucial to improving survival, emphasizing the importance of an adequate and the early source control of infection. Moreover, the pharmacokinetic and pharmacodynamic properties of antibiotics should be considered because of changes in clearance and volume of distribution that are frequently observed in critically ill patients, with the potential to influence the concentration of the drug at the site of infections.As reported in the studies published in this Special Issue, the mechanisms of sepsis may be mainly based on the activation of a hyperinflammatory innate immune system response to microorganisms, including multidrug-resistant pathogens.As a consequence, microorganisms can directly and indirectly stimulate endothelial activation and humoral changes, including the involvement of the innate and the adaptive immune systems.Blood cultures remain the gold-standard diagnostic laboratory technique for the diagnosis of sepsis, but molecular techniques are essential to obtain early microbiological reports.Biomarkers are important tools to differentiate sepsis from noninfectious diseases and thereby contribute to early diagnosis.Prompt empirical broad-spectrum antibiotic therapy and source control of infection are the most effective treatment strategies in sepsis.Pharmacokinetic/pharmacodynamic adjustments are recommended for patients with specific characteristics, including neonates, obesity, burns, and altered renal functions.


Treatment of a wide variety of defects in the oral and maxillofacial regions requires the use of innovative approaches to achieve best outcomes. One of the promising directions is the use of gene-activated materials (GAMs) that represent a combination of tissue engineering and gene therapy. This approach implies that biocompatible materials will be enriched with gene-carrying vectors and implanted into the defect site resulting in transfection of the recipient’s cells and secretion of encoded therapeutic protein in situ. GAMs may be presented in various designs depending on the type of material, encoded protein, vector, and way of connecting the vector and the material. Thus, it is possible to choose the most suitable GAM design for the treatment of a particular pathology. The use of plasmids for delivery of therapeutic genes is of particular interest. In the present review, we aimed to delineate the principle of work and various designs of plasmid-based GAMs and to highlight results of experimental and clinical studies devoted to the treatment of periodontitis, jaw bone defects, teeth avulsion, and other pathologies in the oral and maxillofacial regions. Application of regenerative medicine and tissue engineering methods has great potential for improvement of oral and dental tissue regeneration and can be called regenerative dentistry . RegenerPrevious studies have shown that the use of various growth factors can increase clinical attachment level, reduce gingival recession, and enhance linear bone gain in patients with periodontal osseous defects, and stimulate proliferation of gingival fibroblasts, osteoblasts, periodontal ligament cells, etc. ,6,7,8. HComposition of plasmid-based GAMs implies the presence of the following essential elements: naked plasmid or plasmid/delivery system complex, encoded therapeutic protein, and biocompatible material, enriched with plasmid. The way of attaching plasmid to the material is also of importance since it may influence kinetics of plasmid release. Variability inside GAM elements allows achievement of different effects. Thus, various encoded proteins exert different therapeutic functions. Materials used for GAM creation may also possess different properties depending on chemical structure and biomechanical parameters that may influence tissue regeneration and clinical outcome ,12,13. TThe use of GAM has already shown its potential in pre-clinical studies for the treatment of bone and skin defects ,15. In tThere are various approaches to deliver genetic material including viral and non-viral delivery vectors. It is generally considered that viral vectors have higher transduction efficiency compared to plasmid vectors; however, viral vectors possess several disadvantages, such as immune response. At the same time, plasmids have lower immunogenicity, and their transfection efficiency may be enhanced using special delivery systems . ConsideDespite a relatively small number of published works on GAMs in the field of regenerative dentistry, there is a substantial heterogeneity in GAM design. Various materials, encoded proteins, and delivery systems, as well as methods of attaching plasmid to the material, were used for GAM creation. In the present review, we will discuss works on clinical and pre-clinical use of plasmid-based GAMs in regenerative dentistry and analyze various approaches to their creation.GAMs’ mechanism of action is based on the combination of the properties of the material to form a provisional template for new tissue formation in the area of the defect and the properties of a gene-carrying vector to transfect recipient’s cells and stimulate local secretion of encoded protein. Exact mechanisms of action are deeply dependent on pathology being addressed, type of biocompatible material used, encoded protein, type of delivery system, and approach to attaching plasmid (or a plasmid/delivery system complex) to the material.Transfection of cells may be performed in vitro by culturing cells with non-viral vectors, followed by seeding of cells on the material and implantation into the damaged area . HoweverEvery particular event that occurs upon GAM implantation represents a separate and complex field of extensive research. Next, events that occur upon GAM implantation will be briefly delineated.The technique of GAM positioning at the injury site depends on material and treated pathology. For example, fixation of gene-activated octacalcium phosphate (OCP) within a bone grafting site was achieved via soft tissue suturing in a clinical trial . In vivoAfter implantation, the GAM starts to exert dual functions explained by a combination of tissue engineering and gene therapy in the GAM approach. Aside from being tools for incorporation of the plasmid or plasmid/delivery system complex, the paradigm of tissue engineering implies that materials should represent a provisional template for cell migration, adhesion, proliferation, differentiation, and deposition of extracellular matrix at the defect site . ReleaseTransfection of cells occurs via uptake of naked plasmids of the plasmid/delivery system complex. Some works used naked plasmids for cell transfection in vivo and in vitro, suggesting their ability to be internalized and result in increased protein secretion , or in rAfter entering the cell, free DNA is complexed with intracellular proteins that facilitate microtubule-mediated cytoplasmic transport of plasmid DNA to the surface of the cell nucleus ,29. PlasThe host’s RNA polymerase II facilitates plasmid DNA transcription and mRNA synthesis. The duration of transgene expression greatly depends on the cell type that was transfected. For example, muscle cells are characterized by long-lasting expression in mice for more than a year, presumably because of the absence of cell division . When GATransfected cells start to secrete plasmid-encoded proteins which may be directed to the achievement of various goals. For example, chitosan/collagen matrix enriched with plasmid encoding platelet-derived growth factor (pPDGF) promoted proliferation of human periodontal ligament cells (hPDLCs) and formation of a periodontal connective tissue-like structure in vitro . At the In the field of regenerative dentistry, there is heterogeneity in the design of plasmid-based GAMs. It is explained by the diversity in the following fundamental structural elements of the GAMs: (1) type of material used, (2) type of encoded protein, (3) type of delivery system, (4) method of attaching plasmid or plasmid/delivery system complex to the material. The exact GAM design may be chosen considering the type of treated pathology and individual patient characteristics. Next, we will briefly characterize structural elements that were used for GAM creation in the field of regenerative dentistry.In tissue engineering, a variety of natural and synthetic materials are used. In order to be used for creation of a GAM, material must not only be biocompatible and support tissue regeneration but also be able to attach a naked plasmid or a plasmid/delivery system complex. To create GAMs in the field of regenerative dentistry, various materials are used depending on the scientific and clinical problem being solved.Collagen is a biocompatible biodegradable material capable of stimulating migration of human oral region fibroblasts, endothelial, and periodontal ligament cells , and heaHydroxyapatite is frequently used for bone grafting due to the resemblance of its structure and composition to the natural mineral phase of human bone tissue, and osteoconductive and osteoinductive properties . In dentCoral is a natural porous biocompatible material also used for bone tissue engineering due to its osteoconductive and mechanical properties . Coral sPoly(lactic-co-glycolic acid) is a biocompatible and biodegradable copolymer of lactic and glycolic acid. Depending on the composition and manufacturing conditions, PLGA may be tailored to have a certain degradation and drug/plasmid release rates ,44, mechGelatin is a biocompatible biodegradable natural bio polymer characterized by cost-effectiveness and low immunogenicity, as well as various physical and chemical properties depending on the preparation technique .Octacalcium phosphate (OCP) is a mineral precursor of apatite crystals in bone tissue; it possesses osteoconductive and osteoinductive properties and allows adhesion of bioactive molecules .Titanium and its alloys are widely used in dental prosthetics and for the reconstruction of maxillofacial bone defects as well as in orthopedics ,49. DespThe importance of growth factors in soft and bone tissue regeneration has already been shown in multiple studies. Next, various proteins encoded by plasmids in GAMs used in regenerative dentistry will be briefly overviewed.Vascular endothelial growth factor (VEGF) not only acts as a mitogen for endothelial cells stimulating angiogenesis but also promotes human periodontal ligament stem cells’ (hPDLSCs) odonto-/osteogenic differentiation in vitro ,51. GenePlatelet-derived growth factor is capable of stimulating osteoblast, periodontal ligament cell, and gingival fibroblast proliferation ,8. CliniFibroblast growth factors (FGFs) are a large family of regulatory molecules that have promoting effects on a variety of cells including hPDLCs and hPDLSCs ,51. An iBone morphogenetic proteins are signaling molecules with multiple functions and members of the TGF-β superfamily . BMP-2 iAlthough the benefits of using growth factors have been shown in many studies, their use has some disadvantages, including a short half-life when applied to the defect site and application of supraphysiological doses . The useEnsuring the efficient transfer of genetic material into the cell is one of the main challenges in gene therapy. To enhance plasmid transfection efficiency, a variety of non-viral delivery agents are used. Non-viral delivery systems may be represented by biodegradable and non-biodegradable polymers, lipids, peptides, inorganic materials , hybrid systems , calciumPolyethyleneimine (PEI) is a positively charged cationic polymer that facilitates plasmid entry through negatively charged cell membranes via condensation of negatively charged plasmid DNA . One of Chitosan is a natural cationic biodegradable polymer capable of interacting with negatively charged plasmid DNA via electrostatic forces . Low cytCationic liposomes are widely used gene delivery systems. Cationic liposomes act by binding positively charged head groups of lipids with negatively charged phosphate groups of plasmid DNA and by surrounding DNA molecules .Delivery of naked plasmid DNA is also considered as a promising direction and has some advantages, among which is the absence of delivery systems with potential cytotoxicity. Supercoiling of plasmid DNA also aids more efficient transfection .Various methods of attaching the plasmid or plasmid/delivery system complex to the material are used and are of importance since this parameter may affect the kinetics of plasmid release and duration of transgene expression . MethodsPhysical methods do not imply creation of bonds between plasmid and material and include emulsion electrospinning that allows for the incorporation of the buffer with the target plasmid into the volume of polymeric microfibers for subsequent release in the course of material degradation ,67. ImpoFrom a technical point of view, the attachment of a naked plasmid or a plasmid/delivery system complex to the material may be achieved by different methods, some of which were described by Peng et al. . Among sGAMs of various designs were investigated in vitro, in vivo, and in clinical studies for potential treatment of periodontitis, jaw bone defects, dental pulp capping, gene-activation of titanium implants, improvement of teeth replantation, etc. Despite the growing interest towards gene therapy and the use of GAMs in different fields, the overall number of studies on GAMs in the field of regenerative dentistry is not large .Periodontitis is a multifactorial inflammatory disease of the oral cavity that remains one of the greatest challenges in modern dentistry . This coHuman periodontal ligament cells are mainly presented by fibroblasts and are one of the key players needed for periodontitis treatment since they surround the teeth and attach them to the inner wall of alveolar bone. Impacts of GAMs with various designs on hPDLCs were studied in various in vitro and in vivo studies. An increase in collagen synthesis represents a positive sign since collagen is one of the main proteins of periodontal ligament attaching the tooth to the alveolar bone . In one TGF-β1 gene resulted in greater secretion of TGF-β1, and higher levels of hPDLC proliferation and collagen type I and III mRNA compared to the plasmid-enriched scaffolds [However, it was previously shown that chitosan/collagen scaffolds enriched with a viral vector carrying the caffolds . Higher caffolds . Importacaffolds . These rBMP-2 gene (pBMP-2) and PEI and on single axial electrospun PLGA scaffold with the same plasmid/delivery system complex embedded [Advanced and severe stages of periodontitis are also characterized by alveolar bone destruction, which is one of the main challenges in dentistry . The useembedded . Study rembedded .One of the important tasks in gene therapy is the determination of the optimal time during which the synthesis of the target protein will yield best results. Reparative processes in the body are finely orchestrated and the synthesis of a particular protein can have a positive effect on tissue regeneration if it occurs during a physiologically appropriate time ,83. In tThe need for teeth replantation may arise in cases of traumatic tooth avulsion . ExternaBone defects may appear in the oral and maxillofacial regions due to various reasons, including trauma, tumors, cysts, apicectomies, severe periodontitis, etc. .The first case of clinical use of GAM for the treatment of mandible bone defects was presented by Bozo et al. in 2016 . CollageStudies investigating effects of various plasmid doses are of particular importance. Thus, in the study by Kolk et al., titanium discs were gene-activated with various concentrations of plasmid in copolymer-protected PEI/pBMP-2 polyplexes and coated with poly, and used for the treatment of critical-size mandibular bone defects in a rat model . ImportaWorks comparing effects of recombinant growth factors and plasmids are of utmost value and deserve the closest attention. In continuation of the previous work, Kolk et al. used polylactide-coated titanium discs enriched with either copolymer-protected PEI/pBMP-2 or recombinant human BMP-2 . CompletApplication of novel methods in regenerative dentistry is of particular interest. For example, 3D printing shows great promise for the creation of personalized scaffolds that will precisely match the configuration of the defect ,89. In tTooth extraction may also lead to alveolar bone loss, conditioning the need for restoration of bone and soft tissues . In the Dental implant placement has become a common practice with high success rates. At the same time, avoidance of peri-implant diseases and acceleration of osseointegration still remain clinical tasks of utmost importance .Peri-implant diseases are associated with inflammation of the tissues around the implant and are typically caused by bacterial plaque . Rapid rOsseointegration of titanium implants is one of the prerequisites for successful dental implantation. The titanium surface undergoes various modifications to enhance osseointegration . CoatingOne of the promising directions in GAM creation is a layer-by-layer coating technique that allows the adherence of multiple layers of a plasmid or plasmid/delivery vector complex. Thus, Jiang et al.  showed tDental pulp represents an unmineralized connective tissue comprising peripheral nerves, capillary blood vessels, lymphatic elements, and fibroblastic and stem cells . Pulp caStem/stromal cells were successfully isolated from the gingiva, periodontal ligament, dental pulp of permanent and deciduous teeth, buccal fat pad, dental follicle, and apical papilla of developing teeth . The useThe use of GAMs was also studied in order to assess their impact on DPSC behavior. Chitosan/collagen scaffold enriched with pBMP-7 was shown to stimulate human DPSCs as was seen in the higher values of ALP activity and calcium content in comparison with non-activated materials . Of noteIn vitro studies have shown positive effects of GAMs on behavior of various cells, and animal studies also demonstrated positive results of GAM use for experimental treatment of certain conditions. For example, GAM application led to the reduced root resorption of replanted teeth in dogs and enhanced alveolar bone regeneration after tooth extraction in rats ,17. At tGAMs consist of various elements and may include not only naked plasmids but also plasmids with special delivery systems intended to enhance transfection efficiency. At the same time, the use of particular types of delivery systems may be associated with cytotoxic effects and immune response from the recipient ,103,104.Since regeneration of various tissues is accompanied by finely orchestrated expression of particular genes at certain periods of a regenerative process, it can be concluded that there is a need to clearly select the time points and intervals during which gene expression and secretion of encoded therapeutic proteins will be physiologically useful. As evidenced in the study by Plonka et al., the use of GAM encoding PDGF-B did not result in accelerated regeneration of periodontal alveolar tissues in rats , which mOne of the most valuable properties of GAMs is a provision of spatiotemporal control over plasmid release via choosing appropriate methods of plasmid attachment to the material and parameters of material degradation . The proAmong studied advanced approaches for gingival tissue regeneration is the use of collagen materials . At the A promising direction that deserves further investigation is the creation and the use of delivery systems allowing cell-specific transfection of particular cell types in the oral and maxillofacial regions. This approach may find application during avulsed teeth replantation when stimulation of periodontal ligament regeneration may be achieved with selective transfection of periodontal ligament cells without stimulating osteoblasts for prevention of ankylosis. Selective cell transfection may possibly be achieved with the use of antibody-containing delivery systems ; howeverFurther research is needed to investigate GAMs’ potential to a full extent and to establish the most beneficial GAM designs for regeneration of specific tissues.GAMs are composed of multiple elements and exhibit substantial variability in the design. To achieve the best results, it is critical to carefully select an optimal combination of materials, encoded proteins, plasmid concentrations, and delivery systems in the composition of a GAM directed to regeneration of specific tissue.Plasmid-based GAMs represent valuable tools in regenerative dentistry, allowing for the gradual prolonged secretion of therapeutic proteins. Genetic activation of materials with plasmids is of great interest since plasmids allow for the avoidance of disadvantages of viral vectors. Development of novel materials, delivery systems, plasmids, and methods of attaching genetic constructs to the materials, as well as conduction of new comparative studies, will facilitate further progress.


Impairments in social and non-social cognition are common in psychosis and may be sparsely present even before the onset of the disorder. Genetic and environmental influences have been linked to cognitive dysfunctions, which, in turn, may significantly impact the real-world functioning of people with psychosis. The role of gender in determining the interplay between cognitive skills, risk factors, and outcomes has been relatively unexplored. Nevertheless, identifying putative gender differences in cognitive functions and their correlates may favor the identification of individualized prevention and treatment strategies.None Declared


Soil salinity is a major abiotic stress in global agricultural productivity with an estimated 50% of arable land predicted to become salinized by 2050. Since most domesticated crops are glycophytes, they cannot be cultivated on salt soils. The use of beneficial microorganisms inhabiting the rhizosphere (PGPR) is a promising tool to alleviate salt stress in various crops and represents a strategy to increase agricultural productivity in salt soils. Increasing evidence underlines that PGPR affect plant physiological, biochemical, and molecular responses to salt stress. The mechanisms behind these phenomena include osmotic adjustment, modulation of the plant antioxidant system, ion homeostasis, modulation of the phytohormonal balance, increase in nutrient uptake, and the formation of biofilms. This review focuses on the recent literature regarding the molecular mechanisms that PGPR use to improve plant growth under salinity. In addition, very recent -OMICs approaches were reported, dissecting the role of PGPR in modulating plant genomes and epigenomes, opening up the possibility of combining the high genetic variations of plants with the action of PGPR for the selection of useful plant traits to cope with salt stress conditions. The 21st century can be considered the most complex to face for agriculture, presenting a series of challenges. People must (i) produce more food to feed a growing population, with a reduction in the labor force; (ii) adopt more sustainable and efficient production methods and adapt to climate change; (iii) produce more feedstocks for a potentially huge bioenergy market; and (iv) sustain the development of the many agriculture-dependent developing countries . In addiIn this context, beneficial microorganisms colonizing plant roots represent a promising tool to alleviate salt stress in various crops, thus representing a strategy for increasing agricultural productivity in saline soils .In this paper, we reviewed the recent literature about (i) the roles of rhizosphere beneficial microorganisms, in particular plant-growth-promoting rhizobacteria (PGPR) in alleviating plants’ salt stress; (ii) the molecular mechanisms involved in the cross-talk between plants and PGPR; and (iii) recent -OMICs approaches to understanding molecular PGPR–plant interactions, which will open future perspectives for the application of PGPR in the field and for the microbiome-based selection of salt-tolerant crop genotypes.Since their earliest evolution, land plants have been associated with a complex microbial community that includes beneficial microorganisms. The presence of beneficial microorganisms helped early land plants to respond to the environment and face challenges such as access to nutrients, new and/or stressful conditions, and pathogens ,13.In soil, there is a gradient of intimacy between plant roots and microorganisms, and the plant’s influence on the microbial community is greater near the root surface. A volume of soil specially influenced by plant roots or associated with the roots and plant-produced material is defined as a rhizosphere , a term Among the microbial communities colonizing the different plant regions , the one associated with the roots (rhizobiome) is undoubtedly the most populated and most sophisticated of all microbial communities associated with high plants . The rhiThe root tip senses changes in the concentration of exuded metabolites and translates them into signals to modify root growth. Through root exudate flux, plants can locally enhance concentrations of many common metabolites, which can serve as sensors and integrators of the plant nutritional status and of the nutrient availability for the rhizosphere microbial community ,23. PlanThe microbial population colonizing the rhizosphere includes bacteria, which are the most abundant, as well as fungi, protozoa, and algae, and its composition is markedly different from that of its surrounding soil ,27. RegaThe literature available suggests a two-step selection process that gradually distinguishes the root microbiota from the surrounding soil biomes and determines differences in the dimension and composition of the microbial community. In the first step, it is suggested that rhizodeposition and the recognition of the host cell wall promote organotrophic bacteria growth, thereby initiating the first community shift of the soil biome. In the second step, the microbial subpopulation undergoes a second selection process near and within the roots, based on host genotype-dependent selection, defining the profiles of the microbial communities that prosper in the rhizoplane and within the roots ,30. SimiIn 1978, Kloepper and Schroth  introducAllorhizobium, Azorhizobium, Bradyrhizobium, Mesorhizobium, and Rhizobium of the family Rhizobiaceae . F. F192]. In addition, microbiota abundance in the rhizosphere assessed by metagenomics allowed the mapping of the host genetic determinants of the rhizosphere microbiota in wild and domesticated genotypes of barley , therebyParaburkholderia phytofirmans PsJN into the flowers of soybean and pepper. This method drove the inclusion of the bacterial strain in the progeny seed microbiome, thus inducing vertical inheritance to the offspring generation.Another interesting approach involves the introduction of PGPR into seeds to modify the plant seed microbiome. It involves introducing a microbial strain into the parent plant during flowering . After sMYC, DREB2, and WRKY, extensive research remains to be conducted to understand the signaling pathways involved in plant and PGPR cross-talk, inducing the plant’s salt tolerance response. In addition, the possibility of applying PGPR consortia in the open field for the cultivation of crops in saline soils needs more in-depth studies, to verify the efficacy of PGPR formulations on different plant species or genotypes. Furthermore, the use of -OMICs approaches to explore the highly genetic and epigenetic variations of plants combined with the action of PGPR could be useful for the selection of plant phenotypes able to cope with environmental issues such as salt stress.In conclusion, in many works described in this review, PGPR have been shown to be effective in improving plant growth under salinity. Thanks to these bacteria’s ability to grow under salt stress conditions and their effects on plant growth, they are a very interesting, sustainable solution for improving crop productivity in saline soils. Although some PGPR have been associated with the modulation of genes belonging to different signal transduction pathways such as


The acute inflammatory process is a protective and ideally self-limited response aimed at neutralizing infectious, traumatic, ischemic, toxic and autoimmune aggressions. However, uncontrolled inflammation can be profoundly deleterious. Indeed, in the last two decades, enormous paradigm shifts have occurred with the recognition of inflammation as a central pathophysiological mechanism of several prevalent and harmful diseases and, more recently, with the uncovering of an active resolution of inflammation response mediated by a large family of specialized proresolving mediators (SPM), such as lipoxins, resolvins, protectins and maresins. Since then, new avenues have been opened for the understanding and treatment of a broad range of central and peripheral pathologies, from neurological to cardiovascular disorders. This Research Topic aimed to gather the current knowledge and shed light on the pharmacological role and therapeutic potential of endogenous and exogenous SPM in neurological and cardiovascular diseases, focusing not only on their proresolving and anti-inflammatory effects, but also on their pleiotropic impact. Five papers, including two reviews and three original research articles, were published within this Research Topic.Li et al. focused on the beneficial effects of resolvins in neurological diseases. They first presented a general overview of the endogenous biosynthesis of resolvins and their intracellular signaling pathways, with more specific detail on resolvin receptors and cellular targets in the central nervous system. In the second part, the authors provided evidence from clinical and preclinical studies on the protective role of resolvins in neurodegenerative , neuroimmune  and cerebrovascular disorders (ischemic stroke), ending up with a discussion about their therapeutic potential in neurological disorders and current limitations to their clinical application. A second review paper of Zaninelli et al. approached the pathophysiological role of SPM in arthritis. Authors extensively described SPM classes and presented a structured overview of the current knowledge, according to the precursor lipid substrate: arachidonic acid, eicosapentaenoic acid (EPA) and docosapentaenoic/docosahexaenoic acid (DPA/DHA). Both preclinical and clinical data supporting the contribution of SPM to disease status and potential therapeutic use in arthritis were reviewed.Starting by the review articles, 4 (LXA4) dampening and preeclampsia. The paper by Tao et al. demonstrated the analgesic effect of 1 kHz spinal cord stimulation in an established (2–3 weeks post-induction) rat model of spared nerve injury-induced neuropathic pain. Quite remarkably, authors described decreased central and peripheral levels of the pro-inflammatory mediator IL-1β, and increased central production of the SPM resolvin D1 (RvD1), which support a fundamental role of modulation of neuroinflammation in the analgesic effects of spinal cord stimulation, in neuropathic pain context. Further evidence supporting this hypothesis was provided by an additional experiment reported in the paper, where intrathecal RvD1 administration produced an analgesic pattern similar to that obtained with spinal cord stimulation. Zhao et al. showed the analgesic effect of intrathecal protectin DX (PDX) in a lumbar radicular pain rat model, induced by non-compressive lumbar disc herniation. According to these authors, PDX treatment reverts the pro-/anti-inflammatory imbalance associated with the model, acting through facilitation of the spinal cord autophagy flux, and activation of adenosine monophosphate-activated protein kinase signaling. Lastly, Liu et al. paralleled the study of pregnant women samples and of a rat model of preeclampsia to demonstrate that the induction of this disease by glucocorticoids depends on the reduction of endogenous LXA4 production, thus placing LXA4 insufficiency as a disease mechanism in preeclampsia. Noteworthy, LXA4 exerted several protective pleiotropic effects  in experimental preeclampsia, thus showing a potential for the prevention or treatment of this severe gestational disease associated with multisystemic inflammation and endothelial dysfunction.Regarding the original papers, there were two neurological-related studies addressing the analgesic effects of SPM and one cardiovascular research article investigating the relationship between glucocorticoid exposure, lipoxin ASumming up, pre-clinical and clinical data presented in the articles of this Research Topic align to support not only the importance of SPM in inflammation resolution, but also their usefulness as pharmacological approaches to treat neurological and cardiovascular diseases, including neurodegenerative, neuroimmune and cerebrovascular disorders, several chronic pain syndromes and preeclampsia.Future studies would be important to address sex-related differences in the protective role of SPM and additional pleiotropic effects of these mediators. It would also be useful to expand the knowledge on resolution of inflammation to other diseases of the neuroscience and cardiometabolic areas, such as depression, heart failure and diabetes, given the growing body of evidence supporting the efficacy of SPM precursors in the improvement of these conditions .


Inverse modeling approaches in cardiovascular medicine are a collection of methodologies that can provide non-invasive patient-specific estimations of tissue properties, mechanical loads, and other mechanics-based risk factors using medical imaging as inputs. Its incorporation into clinical practice has the potential to improve diagnosis and treatment planning with low associated risks and costs. These methods have become available for medical applications mainly due to the continuing development of image-based kinematic techniques, the maturity of the associated theories describing cardiovascular function, and recent progress in computer science, modeling, and simulation engineering. Inverse method applications are multidisciplinary, requiring tailored solutions to the available clinical data, pathology of interest, and available computational resources. Herein, we review biomechanical modeling and simulation principles, methods of solving inverse problems, and techniques for image-based kinematic analysis. In the final section, the major advances in inverse modeling of human cardiovascular mechanics since its early development in the early 2000s are reviewed with emphasis on method-specific descriptions, results, and conclusions. We draw selected studies on healthy and diseased hearts, aortas, and pulmonary arteries achieved through the incorporation of tissue mechanics, hemodynamics, and fluid–structure interaction methods paired with patient-specific data acquired with medical imaging in inverse modeling approaches. Given the vast experimental evidence of mechanical factors producing effects on cellular differentiation, signaling, communication, and function –5, in siHowever, for biomechanical models to provide low-risk patient-specific solutions, personalized non-invasive clinical studies must be readily available to quantify regional cardiovascular function. Current medical imaging technology, namely echocardiography and magnetic resonance imaging (MRI), offers not only anatomical information but also high-resolution kinematics data of tissue motion and blood flow –12. KineThe inverse method, or data assimilation method, is an approach that allows solving classic mechanics problems “backwards”; that is, retrieving material properties and dynamic information (stress and forces) using measured kinematic information and loading boundary conditions as input . SeveralThe relevance of patient-specific modeling and its potential impact on the future of personalized and predictive health care has been acknowledged by several funding agencies. In 2003, the Interagency Modeling and Analysis Group was formed from the collaboration of nine institutes of the National Institutes of Health (NIH) and three directorates of the National Science Foundation (NSF). This group released its first funding opportunity in 2004 under the title “Interagency Opportunities in Multiscale Modeling in Biomedical, Biological, and Behavioral Systems Solicitation”, which has been regularly reissued since then, and led to the creation of the Multiscale Modeling Consortium which includes over 100 projects on multiscale modeling of biological systems. The European Union initiated the “Structuring the Europhysiome” Consortium in 2006, which led to the Virtual Physiological Human project, an ongoing initiative that aims to bring together policymakers, regulatory agencies, funding bodies, industry, and research organizations towards the development of integrated computer models of the mechanical, physical, and biochemical functions of the living human body . These iInverse modeling of biomechanical systems requires the confluence of state-of-the-art techniques from several disciplines including clinical care, medical imaging, simulation engineering, data analysis, and computer science . InverseInverse modeling analyses can also be applied to in vitro experimental setups. The main advantage of this approach is that input data is not limited by the available clinical tests, mechanical loads can be precisely controlled, and kinematics can be measured with high-resolution instruments. Furthermore, the target of the inverse method can be defined not only in terms of kinematic information but also in terms of controlled mechanical loads and stress measurements . MoreoveThis article aims first to serve as a referential document for concepts and methods from all involved disciplines on patient-specific in vivo inverse modeling; and secondly, to highlight the potential clinical application of patient-specific inverse modeling in the cardiovascular research field. Specifically, we review the fundamentals of cardiovascular tissue and blood biomechanics, modeling and simulation, and medical imaging, as they relate to the inverse modeling approach and its applications in cardiovascular medicine. In 2.Modern biomechanics consists of the formulation of governing equations describing balances of mass, linear and angular momentum, and energy to biological systems and physiological processes. The human body maintains a uniform and stable temperature through homeostatic thermoregulation. Thus, contributions due to temperature change in the internal energy of the material, heat fluxes, and heat supply are typically negligible to the energy balance, which is in turn reduced to the balance between deformational energy and stress power (thermodynamic work). In classical continuum mechanics of purely mechanical processes, the balance of angular momentum directly translates to the symmetry of the stress tensor, and therefore, the relevant governing equations for most cardiovascular mechanics applications consist only of the balances of linear momentum and mass. However, most biological systems are open, and continuously interact with their surroundings, and thus the conservation principles must be handled carefully, especially with respect to tissue growth and atrophy within relevant timescales.Given that the resolution of most in vivo medical imaging is on the scale of millimeters, only phenomena occurring at the tissue level can be directly associated with these measurements. The assumption of material continuity is reasonable for the formulation of the governing principles at this scale, leaving any additional considerations dealing with the extracellular and intracellular micro-environments to be included ad hoc with additional modeling formulations and constitutive equations.To apply these principles, it is necessary to relate the stress tensor to kinematic measures, which is in essence the description of the mechanical behavior of the material under study. This information is provided by a constitutive equation; these can be either phenomenological equations “arbitrarily” formulated to reproduce experimental observations, or analytical expressions inspired by theoretical interactions of the material constituents at the micro or molecular scale. The selection of adequate models to describe the phenomena of interest is key to the success of any engineering analysis. The selected model must be complex enough to describe the most salient observable features at the scale of interest, while ideally being simple enough to provide a rational interpretation of its parameters and results and render a computationally tractable numerical problem. After fitting these parametrized models to experimental data, the constitutive equation can provide an additional understanding of the underlying mechanisms associated with the mechanical response of the material. Models of increased complexity usually require a larger number of parameters to be fitted, and overparametrized models can lead to solution multiplicity which obscures its interpretation and validity. For the sake of generality, the constitutive equation must also be independent of the frame of reference, comply with the second principle of thermodynamics, and yield amenable mathematical treatment and systems of equations that are solvable .After formulating the constitutive relation as a function of specific unknowns , the resulting system of governing equations is then particularized into specific problems by the definition of temporal and spatial domains of interest and the imposition of appropriate boundary and initial conditions. To obtain a unique solution, it is necessary to constrain the problem by assigning a first-order boundary condition on at least one of the boundaries . On the rest of the boundaries, higher-order boundary conditions can be applied to impose distributed forces such as a known pressure. Once solved, the result of this forward formulation is the transient spatial distribution of displacement or velocity throughout the domain as a result of the specified loads and material properties. From this kinematic information, strain and strain-rate distributions can be numerically derived, and the stress distributions retrieved via the constitutive equation.In the following subsections, we present a review of the main features of the mechanical behavior and composition of cardiovascular tissues and blood, as well as the constitutive equations that have been developed and applied to model such behaviors. Then, we review the algorithms implemented to model fluid–structure interactions and their importance in cardiovascular mechanics simulations. Finally, we briefly discuss the modeling of biological tissue growth and remodeling by the application of the constrained mixture theory.2.1.The study of soft biological tissues under the framework of finite elasticity was initiated by Y.C. Fung and others in the late 1960s, setting the basis of modern biomechanics ,26. As iCardiovascular tissues are comprised of multiple layers of cells and extracellular matrix (ECM) components. The ECM is a network of macromolecules that is continuously synthesized and degraded by active cells and functionally provides them with structural and biochemical support. Typically, collagen, elastin, and fibrillin are regarded as the main structural constituents responsible for the macroscopic mechanical behavior of cardiovascular tissues . HealthyAdditionally, cardiovascular tissue is muscular in nature and actively contracts/distends. Thus, its mechanical behavior is affected by the activation of actin-myosin sliding filaments, which depends on ion-based chemical signaling and determines the muscular tone. In the myocardium, striated muscle activation is responsible for cardiac contraction. In large arteries, contraction of smooth muscle cells regulates downstream vascular resistance, blood flow, and propagation of the pressure-pulse wave along the vascular tree.Of note, cardiovascular tissue also exhibits viscoelastic behavior, which has been established with stress relaxation, creep, and strain-rate experiments. It has been argued that viscous energy dissipation of healthy tissue, functioning at a regular physiological rate (~1 Hz), is negligible compared to stored strain energy . Neverth2.1.1.Linearized elasticity falls short in describing the complex behavior of biological tissue, not only because the mechanical response is highly non-linear but also the material undergoes finite motions and deformations. In the case of non-linear behavior, it is usually convenient to employ the formulation of hyperelasticity and express the constitutive equations as the relation of a scalar stored energy density function to the deformation gradient tensor or the strain tensor . The scalar energy density function represents the amount of deformational energy stored per unit volume and is defined in such a way that the stress tensors can be obtained from their derivatives with respect to the strain or stretch tensor.The passive behavior of cardiovascular tissue is characterized by an increasing resistance to deformation with strain. This behavior is represented by an increasing slope in the stress versus strain/stretch and strain energy versus strain/stretch curves, with a close to zero slope at zero strain and rapidly increasing at physiological ranges. This behavior has been attributed to the structural characteristics of the ECM components. It has been suggested that the increasing resistance to deformation with strain is owed to the progressive engagement of wavy bundles of elastin and collagen fibers to support the mechanical loads .Hyperelastic isotropic material models, such as the Neo–Hookean and the Mooney–Rivlin constitutive equations, can accurately describe the behavior of amorphous bodies such as lipid pools in atherosclerotic formations, and fit some portions of the pressure–volume relation of blood vessels. The symmetry of these material models allows its representation as to the linear combination of the deformation gradient invariants weighted by the material properties. This formulation allows the determination of unique material properties for a given mechanical behavior. However, these models fail to reproduce the characteristic highly nonlinear and anisotropic behavior of cardiovascular tissue  and 3. TIn the past decades, many microstructure-inspired constitutive models have been proposed to specifically suit cardiovascular tissue behavior ,41. FibeAdequate modeling of active contraction is key for an accurate description of cardiovascular function in general, being particularly critical for modeling the heart. Conceptually, there are two possible approaches, the active stress models are the most common and they assume the stress tensor can be decomposed as the sum of a passive and active component, while active strain models assume a product decomposition of the deformation gradient.2+) to troponin C and its release, tropomyosin movement kinetics, the myofiber length, and the kinetics of cross-bridge tension build-up under perturbation of myofilament length. In practice, the detailed information required for the evaluation of this model is out of reach, so simplified models assume the active stress acts mostly lengthways the direction of myofibers, with a magnitude that is proportional to the fiber length and the activation status. The activation status is often expressed as a time-dependent spatially heterogeneous function ranging from 0 to 1 ..262].One of the earliest inverse analyses of biventricular models with region-wise material heterogeneity was proposed by Marchesseau et al. . The stuIn 2017, Gao et al. performed an inverse analysis on 27 healthy subjects and 11 patients with acute myocardial infarction . GadolinIn 2018, Finsberg et al. compared the LV contraction between healthy adult volunteers and patients with blocked or delayed electrical activation impulses, a condition called left bundle branch block (LBBB) . The stuZhang et al. studied the local effect of ischemia with the segment-wise heterogeneity approach . Five paOne of the main limitations of the above studies is the assumption of either spatial material homogeneity or segment-wise heterogeneity, however, material properties are likely to vary continuously throughout the myocardium. To address this, Balaban et al. proposed an iterative inverse method to resolve the heterogeneous distribution of mechanical properties on an LV model from a 64-year-old heart with systolic heart failure, LBBB, coronary artery disease, and chronic infarction in the inferior section of the LV . 4D echo6.3.Each one of the chambers of the heart is equipped with a discharge valve to ensure unidirectional blood flow, acting mostly passively to changes in transvalvular pressures. The atrioventricular valves are the mitral and tricuspid, for the left and right sides of the heart respectively. These valves typically define the basal plane and separate the atria from the ventricles . They arHeart valve disease is mostly related to regurgitation, stenosis, and atresia. The former consists of backflow due to deficient closing, stenosis is the hardening and thickening of the leaflets, preventing the valve to open properly and result in increased load in the heart, while the latter is a congenital disease where the heart valve is partially or completely absent. Heart valve malfunction can lead to several complications such as heart failure, blood clotting, stroke, and death. Heart valve disease is most common on the left side, as the aortic and mitral valves are loaded with larger pressures, and in consequence, they have received more attention from the medical and scientific community. However, attention to right heart valves has significantly grown in the last two decades along with the awareness of pulmonary artery diseases .There is a considerable body of research on the forward modeling of heart valve function accounting for structural and FSI mechanics, usually validated against in vitro experiments . HoweverIn vivo inverse modeling of ovine heart valves function has been achieved by the use of fluoroscopic markers implanted on the surface of mitral valve leaflets ,276, a tAggarwal et al. estimated the residual strain on human aortic valves by combining in vivo imaging with measurements on explanted tissues . The autThe work of Aly et al. stands out as one of the few in vivo works on human heart valves for the generation of transient anatomical models . In this6.4.Changes in mechanical properties of arterial walls have been associated with the onset of multiple cardiovascular pathologies  and remains an important predictor of cardiovascular morbidity and mortality in clinical practice. This motivated the development of early techniques for the non-invasive assessment of arterial stiffness through the evaluation of luminal area change and pulse wave velocity. These techniques, although useful, can only provide a gross estimation of material properties as they introduce many assumptions and simplifications related to homogeneity, perivascular support, and linearized behavior.The image-based resolution of vascular tissue kinematics is technically challenging; the main reason being the relative thinness of vascular walls. For example, the ascending aorta has a typical thickness of about 2.5 mm, which decreases to about 1.5 mm at the abdominal aorta, and the pulmonary artery is only about 0.2 mm thick. These length scales are comparable to the highest resolutions available on imaging techniques, for which luminal area changes (either with or without contrast agents) remained the main input for early inverse analyses of arteries. However, recent developments in ultrasound speckle tracking and DENSE MRI techniques make available arterial wall displacement measurements on a meaningful number of pixels. Most approaches rely on FEM for the solution of the forward problem  to study the strain distribution in two abdominal aortic aneurysms (AAA) of two adults and the abdominal aorta of six healthy controls . ResultsKrishnan et al., performed inverse model analyses on ascending thoracic aorta aneurysms (aTAA) . These aLiu et al. explored new methods to reduce the computational cost of inverse analyses while studying the mechanical properties of aTAAs. First, they investigated a method based on the computation of wall stress by solving a simplified statically determinate problem to obtain an “almost true” stress field . They coAll these studies assumed material homogeneity of the aortic wall, which is a major limitation for the study of aneurysms. In vitro mechanical tests and histology analyses have been performed on aneurysms from human cadavers revealing both structural and mechanical heterogeneity ,295. Far6.4.2.Atherosclerosis is a chronic inflammatory disease that manifests as the hardening and occlusion of arteries due to the build-up of plaque on the lumen of the arterial wall. Atherosclerotic plaque is a mixture of fatty substances, cholesterol, calcium, and cellular waste, usually enclosed in a fibrous cap. Atherosclerotic lesions are generated at specific regions of the arterial tree, mostly in the vicinity of branch points, the outer wall of bifurcations, and the inner wall of curves . Among mThe main limitation of previous studies is that the current resolution of non-invasive imaging techniques is insufficient to resolve the displacement of atheroma plaques in small vessels such as the carotid artery. To overcome this, Maso Talou et al. utilized intravascular ultrasound technology . This wo6.5.In general, computational modeling of hemodynamics is more resource-consuming than tissue mechanics, as simulations need to account for transient effects and deal with the difficulties introduced by the non-linearities of convection and momentum dissipation. This makes the application of inverse modeling to hemodynamics a challenging task.The use of simplified 0D (lumped) and 1D models can significantly reduce the computational cost. These simplified models have been used on a patient-specific basis and implemented onto inverse modeling approaches to provide useful systemic information about flow distribution, vascular resistance, and the systemic effect of drug treatments ,298. HowTo deal with the computational expense of the forward problem on inverse hemodynamics, Lassila et al. proposed a method for parametrizing the Navier–Stokes equations and patient-specific geometries to reduce the basis of the partial differential equations. The parameterized model is iteratively solved until the algorithm is close to the final solution. At this point, the inverse method then switches to the solution of the full-forward problem using FVM. This method was tested using deterministic and Bayesian optimization algorithms showing promising results on the solution of test cases involving rigid-wall and FSI simulations . Herein,6.5.1.Romarowski et al. applied an iterative inverse method for the hemodynamic study of three descending thoracic aortic aneurysms. CT scans were used to build the 3D models that included the ascending and descending aorta . PC MRI Rispoli et al. proposed a modification to the implementation of FVM for fluid dynamics problems, to introduce the minimization of simulated nodal velocity components to 4D flow data measurements in the linearized SIMPLER algorithm . The met6.5.2.Fluid–structure interaction simulation is itself a complex, resource-consuming process, and its incorporation with inverse models is challenging. Some of the early work by Moireau, Chapelle, D’Elia, Perego, among others, set the bases for inverse modeling of FSI by calibrating models to in vitro experiments and synthetic datasets –303.In 2014, Bertoglio et al. proposed the use of Kalman Filters to estimate the material properties of several regions of the aorta from inverse FSI . AvailabZambrano et al. proposed an iterative inverse method for the study of the pulmonary artery . IntravaThe arterial wall was assumed homogeneous and isotropic linear elastic throughout the entire domain. The fluid–structure interactions were modeled with the coupled momentum method. Boundary conditions consisted of PC MRI-derived inlet flow and three-parameter Windkessel models in the outlets. The elastic modulus of the wall and Windkessel boundary parameters were calibrated by iterating in two nested loops. In the inner loop, the Windkessel parameters were adjusted until the error to the measured pressure waveform is minimized, while the outer loop adjusted the elastic modulus until the error to the measured pressure-area curve is minimized . On each6.6.In 7.Inverse modeling is an analysis tool that can provide detailed information about domain properties and loading conditions using kinematic measurements as inputs. When applied to collected data from controlled in vitro experiments it can provide dynamic information with high levels of accuracy and reliability. In biomedical research, inverse modeling has been coupled with microscope-based imaging techniques to yield relevant information on the response of cardiovascular and engineered tissue to mechanical stimuli at the cellular level. These contributions hold relevant scientific value in the fields of mechanobiology and tissue engineering, however, the extrapolation of these results to patient-specific cases is limited.There is great interest in the development of reliable patient-specific non-invasive medical tools to assess the onset and progression of cardiovascular disease. This has led to significant advances in non-invasive medical imaging, including improvements in resolution, scan time, operational costs, availability, and the ability to quantify detailed regional kinematic information. Inverse biomechanical analyses can exploit this available clinical data to provide patient-specific estimations of dynamic parameters that otherwise require invasive  procedures, such as vascular catheterization, or cannot be measured at all. Inverse modeling fits dynamical unknowns to kinematic data, which would be simply assumed with fixed values on classical forward modeling approaches. However, inverse modeling cannot entirely substitute measurements of absolute pressure (required to define the loading boundary conditions); instead, this technique can be used to estimate other relevant biomarkers defined in terms of pressure or load differences, such as vascular flow resistance. As highlighted in this review of the clinical applications of these methodologies, inverse analyses can estimate stiffness for healthy and diseased cardiac and vascular tissues, identify and delineate pathological lesions, resolve tissue composition, and quantify mechanical loads and stresses during in vivo function. Inverse modeling can also provide physiological rationales for empirically derived risk factors, such as aneurysmal diameter and ventricular volume, as well as yield new sets of physiologically meaningful risk markers. In addition, inverse modeling can deliver insights into how biological tissues respond and adapt to pathology and/or therapies through comparative studies, such as regional changes in active contraction within infarcted hearts or tissue growth and remodeling in aneurysmal arteries.Despite all these advantages, the incorporation of patient-specific inverse-modeling in clinical practice still faces several challenges, including the presence of multiple solutions, uncertainty regarding patient-specific stress-free reference configurations, computational costs, and the lack of required clinical and imaging data. The multiplicity of solutions is a common challenge to any inverse problem, and the solution set can be reduced by constraining the optimization parameters within ranges of expected values, incorporating regularization functionals, sampling stochastic parameters, designing special optimization target functions, and, for the specific case of Bayesian approaches, providing probability distributions of parameters from previous experiences.A step towards resolving patient-specific stress-free references for tissue mechanics is the inverse solution of unloaded configurations through direct and iterative methods. However, it is generally accepted that unloaded blood vessels are not truly stress-free due to the existence of residual stress/strains which are influenced by the heterogeneous growth and continuous remodeling of the tissue, including the prestretch of key extracellular matrix components such as collagen. This issue could potentially be addressed by the implementation of a constrained mixture theory and the in vivo resolution of tissue microstructure via medical imaging.The computational cost of iterative inverse methods is often addressed by simplifications of the forward problem, the use of surrogate models for early optimization stages, utilization of more efficient iterative optimization methods, and the use of parallel computing. Furthermore, the ongoing increase of computational power may allow the solution of complex problems that escapes the reach of current technology.Similarly, it is reasonable to expect that medical imaging technology will continue to evolve, making them more readily available in healthcare practice. The development of data-driven techniques for the support of clinical decision making and treatment planning could also motivate the implementation of image-based kinematics in routine health care.Inverse modeling is just one of many patient-specific techniques that have been proposed as a useful support for clinical practice. Machine learning has been increasingly explored in the last two decades for incorporation into the new field of precision medicine . This teAn additional advantage of simulation-based techniques is their predictive capabilities. Founded on physical and physiological principles, patient-specific inverse problems can be coupled to mechanobiology-inspired growth and remodeling models to potentially predict the progression of diseases and/or the effect of treatments . In conc