Limitations to the Clinical Relevance of Preclinical Studies
\r\n\tThis book intends to provide the reader with a comprehensive overview of the current state-of-the-art on Theileriosis, Babesiosis, and Anaplasmosis, both in humans and domestic animals. Particularly this book aims to permit the researchers to enter into a critical focus on the biology of the parasites, eco-epidemiology of the diseases, clinical manifestations, risk factors, immunology, surveillance, diagnosis, identification, and management of risks as well as the potential economic impact on animal production.
",isbn:"978-1-80356-384-8",printIsbn:"978-1-80356-383-1",pdfIsbn:"978-1-80356-385-5",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,isSalesforceBook:!1,isNomenclature:!1,hash:"3d72ae651ee2a04b2368bf798a3183ca",bookSignature:"Prof. Elisa Pieragostini",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11577.jpg",keywords:"Zoonosis, Global Climate Change, Epidemiology, Risk Factors, Haemoparasites, Prevention & Control, Piroplasmosis, Anaplasmosis, Babesiosis, Theileriosis, Tick-Borne Diseases, Ticks",numberOfDownloads:null,numberOfWosCitations:0,numberOfCrossrefCitations:null,numberOfDimensionsCitations:null,numberOfTotalCitations:null,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"February 23rd 2022",dateEndSecondStepPublish:"April 29th 2022",dateEndThirdStepPublish:"June 28th 2022",dateEndFourthStepPublish:"September 16th 2022",dateEndFifthStepPublish:"November 15th 2022",dateConfirmationOfParticipation:null,remainingDaysToSecondStep:"3 months",secondStepPassed:!0,areRegistrationsClosed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Professor of Animal genetics and breeding at Bari University, a researcher in animal genetics related to the resilience of Apulian livestock to enzootic tick-borne haemoparasites and to the involved functional effect of hemoglobin variants. She is a member of national and international scientific associations.",coeditorOneBiosketch:null,coeditorTwoBiosketch:null,coeditorThreeBiosketch:null,coeditorFourBiosketch:null,coeditorFiveBiosketch:null,editors:[{id:"51521",title:"Prof.",name:"Elisa",middleName:null,surname:"Pieragostini",slug:"elisa-pieragostini",fullName:"Elisa Pieragostini",profilePictureURL:"https://mts.intechopen.com/storage/users/51521/images/system/51521.jpg",biography:"Dr. Pieragostini has a Degree in Biological Sciences, DPhil in Biochemistry received from Bologna University, Italy.\r\nShe was an Assistant professor of “Genetics” at the Faculty of Science of Bologna University, a Full professor of “Animal genetics and breeding” Bari University, Italy, and Elected President of course on “Agricultural Science and Technology” (2001-2010) Bari University.\r\nShe retired in 2015.\r\nFrom 1977 to 1984 she carried out intensive work on Drosophila investigating quantitative characters associated with biochemical markers. From the1985 onwards, she had been studying Apulian farm animals focusing on blood-based markers as well as on the structure-function relationship of the different hemoglobin variants come across and their relationship with the Apulian livestock resilience to the enzootic tick-borne diseases. \r\nShe was Project Director\r\nShe is a member of national and international scientific associations.",institutionString:"Independent Researcher",position:null,outsideEditionCount:0,totalCites:0,totalAuthoredChapters:"1",totalChapterViews:"0",totalEditedBooks:"0",institution:{name:"University of Bari Aldo Moro",institutionURL:null,country:{name:"Italy"}}}],coeditorOne:null,coeditorTwo:null,coeditorThree:null,coeditorFour:null,coeditorFive:null,topics:[{id:"16",title:"Medicine",slug:"medicine"}],chapters:null,productType:{id:"1",title:"Edited Volume",chapterContentType:"chapter",authoredCaption:"Edited by"},personalPublishingAssistant:{id:"280415",firstName:"Josip",lastName:"Knapic",middleName:null,title:"Mr.",imageUrl:"https://mts.intechopen.com/storage/users/280415/images/8050_n.jpg",email:"josip@intechopen.com",biography:"As an Author Service Manager my responsibilities include monitoring and facilitating all publishing activities for authors and editors. From chapter submission and review, to approval and revision, copy-editing and design, until final publication, I work closely with authors and editors to ensure a simple and easy publishing process. I maintain constant and effective communication with authors, editors and reviewers, which allows for a level of personal support that enables contributors to fully commit and concentrate on the chapters they are writing, editing, or reviewing. I assist authors in the preparation of their full chapter submissions and track important deadlines and ensure they are met. I help to coordinate internal processes such as linguistic review, and monitor the technical aspects of the process. As an ASM I am also involved in the acquisition of editors. 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Severe sepsis, defined as sepsis with at least one dysfunctional organ, is the leading cause of death in non-coronary intensive care units and is associated with mortality rates of 30-50%. The development of experimental sepsis models to elucidate the progression and pathophysiology of clinical sepsis spans the past eight decades. Studies utilizing models of intra-abdominal sepsis began in the 1930’s with the isolation of endotoxin and the intravenous or peritoneal infusion of live organisms, a model which dominated sepsis research for over 30 years. In the 1960’s, a transition was made from endotoxemia models to a focus on bacteremia. Such models include the injection of live bacteria (Pseudomonas aeruginosa, Escherichia coli, and Staphylococcus aureus), inoculation of feces, and intramuscular, intraperitoneal and subdermal implantation of feces-containing capsules and sponges. Following the use of endotoxemia and bacteremia models, various models involving ischemia and bowel perforation were developed. These models led to the development of the most frequently used sepsis model today, cecal ligation puncture (CLP) and more recently, the colon ascendens stent peritonitis (CASP) model. Over the years several large animal models of sepsis have also been developed, of which canine, ovine, bovine, rabbit, and non-human primates have proven to be most useful. However, the translation of findings and inferences from animal sepsis models to human sepsis remains a challenge. In this chapter, we will provide an overview of experimental models of sepsis, with focus on the merits and limitations of each model. We will also focus on strategies that may improve the translation of results from animal studies to human sepsis. This requires consideration of the limitations of current sepsis models including supportive therapies, considering age, gender, obesity, and exposure to pathogens in animals used, and addressing the risk of bias in experimental sepsis models.
Sepsis is a serious, complicated, heterogeneous condition involving a dysregulated host response to an initial infection and subsequent hemodynamic, cardiovascular, respiratory, metabolic, hormonal, inflammatory, innate and adaptive immune changes [1;2]. Experimental sepsis models have been utilized extensively and developed over the last eight decades to study the progression and pathophysiology of clinical sepsis [3-5]. Intra-abdominal sepsis models were introduced in the 1930s with the isolation of endotoxins and the intravenous or peritoneal infusion of live organisms, models which dominated sepsis research for thirty years. Upon the discovery of new antimicrobial compounds, treatment of intra-abdominal abscesses became a clinical focus in the 1960s and a transition was made from the endotoxin model to the bacterial model of infection [4]. These models used fecal pellets and the addition of an adjuvant to induce infection with the goal of studying peritonitis rather than isolated abscess formation [3]. However, these models fell short as mortality rates relied heavily on bacterial counts and bacterial composition which varied widely between both species and individual subjects. Additionally, the bacterial overload at non-physiologically relevant levels and lack of a sterile environment contributed to a much greater mortality rate in experimental sepsis compared to what was observed clinically [4].
To overcome these limitations, the introduction of defined bacterial inoculum models in the 1970s improved sepsis modeling immensely. These models were also used in landmark studies which uncovered the two-stage nature of intra-abdominal sepsis with gram-negative enteric bacteria inducing the peritonitis phase and anaerobic bacteria responsible for subsequent abscess formation [3]. Antibiotic agents tested using defined bacterial models were successful in either improving survival or reducing abscess formation but were not successful in improving both criteria. This led to the development of models which more closely resembled the progression clinical sepsis.
In the ‘60s and ‘70s, the Clowes group and Wright group were able to create septic conditions in dogs by ligating the cecum at the ileocecal valve, successfully demonstrating both the initial hyperdynamic phase and later hypodynamic phase of sepsis, but neither groups documented bacterial cultures to validate dissemination of pathogens in the peritoneum or circulation [6-9]. A small animal model was later introduced by Ryan
The CLP model of intra-abdominal sepsis was introduced by Wichterman, Baue, and Chaudry in 1980. The group published an insightful review of previous models and introduced a novel sepsis model still widely regarded as the gold standard for modelling polymicrobial sepsis today—the cecal ligation and puncture (CLP) model. Rats were fasted, their cecum was ligated distal to the ileocecal valve, the antimesenteric cecal surface was punctured twice with an 18G ½ needle, and received subcutaneous saline post-operatively. This model induced polymicrobial infection (blood cultures positive for
Multiple aspects of the CLP procedure address the complex, of the clinical course of sepsis. CLP induces polymicrobial infection of the peritoneum with a localized infectious focus, release of bacteria and endotoxic molecular components of pathogens (pathogen-associated molecular patterns or PAMPs) into normally sterile areas in the host, and subsequent translocation of enteric bacteria into the bloodstream, modelling the stages of intra-abdominal clinical sepsis [3]. Under anaesthesia, the cecum is exposed and trauma is induced via a midline skin laparotomy and blunt dissection of the peritoneum to exteriorize the cecum. Avoiding damage to the mesenteric vessels, the cecum is ligated with suture distal to the ileocecal valve, punctured once or twice (through-and-though) from the mesenteric to anti-mesenteric direction halfway between the ligation and cecal end, and aspirated for trapped gasses [3]. A small amount of fecal content is extruded to allow for patency of the puncture(s) and continuous flow of feces post-operatively. The cecum is returned into the peritoneal cavity taking care not to spread fecal content on the incision and the peritoneum and abdomen are closed separately with sutures [11].
Significant elements of the host response to CLP-induced polymicrobial sepsis are present in clinical sepsis. The hemodynamic profiles, cardiovascular response, metabolic phases, systemic involvement of cytokine responses (ex. profiles of interleukins), changes in the innate and adaptive immune response, and abnormalities in mediators of coagulation which occur following CLP are also observed in the clinical course of sepsis [12]. Moreover, CLP involves multiple, complicated elements which are unaccounted for by models of endotoxemia and bacterial inoculum. These include a laparotomy which mimics surgery-induced trauma in the septic patient, the presence of inflamed tissue (peritonitis), necrosis via ligation of the cecal end, apoptosis of specific leukocytes, bacteremia induced by pathogens from a host-derived flora (fecal spillage), and translocation of enteric, living, multiplying bacteria into the bloodstream [12]. The inclusion of these elements as part of the CLP model improves the clinical relevance of outcomes from these preclinical studies.
As observed in clinical sepsis, the hyperinflammatory state which occurs during the systemic inflammatory response syndrome (SIRS) transitions to an immunosuppressed state characterized by a compensatory acute response (CARS) [12;13]. However, the point at which this transition occurs in both clinical and experimental sepsis is unclear. The early hyperdynamic stage of sepsis and the later hypodynamic state following the CLP procedure is indicated by changes in response to immune challenge and changes in peripheral blood cells, plasma levels of cytokines, and chemokines. Neutropenia and lymphopenia are characterized by significant peripheral blood alterations rapidly following CLP parallel to leukocytosis or leukocytopenia observed in the clinical SIRS condition [1;12-14]. Following CLP, total white blood cells, polymorphonuclear cells, and lymphocytes increase rapidly within the first 2 hours, decrease from 2-4 hours, and plateau until endpoint at 8 hours following CLP [15]. The pro-inflammatory response is also characterized by significant increases in cytokines TNFα and IL-6 and chemokines KC, MIP-2, and MCP-1 from non-detectable plasma levels that increase and remain elevated over an 8 hour period [12;15]. Several studies have demonstrated the importance of an early pro-inflammatory response in the progression of sepsis. Antibody-mediated blockade of IL-6 [16;17], complement factor C5a or C5a receptor [18-20], and depletion of neutrophils offered protective effects and increased survival of animals subjected to CLP.
While anti-inflammatory proteins like IL-10 and glucocorticoids are crucial for dampening and terminating the inflammatory response [21-23], the hypoinflammatory phase of experimental sepsis characterized by neutrophil paralysis (shutting down of signaling pathways), apoptosis of lymphocytes and dendritic cells, and elevations in anti-inflammatory mediators significantly increase the susceptibility of the septic host to nosocomial infection. It is during this hypoinflammatory, immunosuppressed state when most mortality is observed in clinical sepsis [21] however it is unknown if this holds true in animal sepsis.
Multiple elements of the CLP procedure can be modified to model the wide spectrum of conditions observed in clinical sepsis. These factors include the number of cecal punctures, gauge of needle used to puncture the cecum, and the length of cecum ligated in the animal [3]. However, there are conflicting findings as to whether the number of cecal punctures affects disease severity. One group reported that two cecal punctures does not result in a significant increase in mortality but is associated with a decrease time to endpoint [24]. Variations in the CLP protocol can be used to produce different disease severities and mimic various stages of the sepsis spectrum from the rapid onset of a robust, hyperinflammatory state to a gradual progression of severe sepsis to an immunosuppression in septic shock. For instance, modifications such as using a smaller gauge/thicker needle [18G ½ rather than a 26G½) or ligating a larger amount of cecum (by placing the suture proximal to the ileocecal valve) can increase the severity and produce a mortality rate that may be more clinically relevant [25].
The flexibility in modeling various severities of disease, ability to recreate hemodynamic, metabolic, and immune changes, the inclusion of surgical trauma, necrosis, and apoptosis of specific cell types which more closely correlate with clinical sepsis contribute to the acceptance of CLP as the gold standard for modeling polymicrobial, intra-abdominal sepsis.
Almost two decades following the introduction of the CLP model by Chaudry et al., Zantl et al. introduced a polymicrobial, peritonitis sepsis model termed colon ascendens stent peritonitis or CASP [26]. The CASP model is a reproducible model suitable for studying the pathophysiology of abdominal sepsis and can be successfully varied by using stents of different diameters ranging from 14G to 22G [26;27]. It can also be used to study surgical interventions which involve the elimination of the infectious focus by stent removal [27]. The prevalence of the CASP model in sepsis studies has only recently increased. Currently, the number of studies utilizing the CLP sepsis model far exceed those using CASP [25].
In the CASP procedure, a laparotomy is performed to exteriorize the cecum, terminal ileum, and ascending colon. The ascending colon wall is pierced and the suture is fixed on the colon wall 15 mm distal from the ileocecal valve. A prepared stent or cannula is used to puncture the ascending colon around 1-2 mm proximal from the suture and the cannula is inserted into the colon and sutured securely. Fecal content is milked through the stent which provides a pathway between the intestinal lumen and the peritoneum, allowing unobstructed influx of enteric bacteria into the peritoneal cavity [26;28]. Disease severity of this model, can be modified by adjusting the size of the cannula used for stenting from 14 G to 20 G [100% lethality and less than 50% mortality at 48 hours following surgery, respectively) [24;28]. Within 3 hours of stent implantation, levels of circulating and systemic cytokines and chemokines including TNFα, IFN-γ, IL-1, IL-12, IL-18, KC/GRO-α, MCP-1, and anti-inflammatory IL-10 increase [26;29;30]. Unlike CLP however, progression of sepsis in the CASP model appears less dependent on the initial immune response elicited by TNFα and more heavily focused on innate immune activation via toll-like receptors (TLRs) and TLR signalling. In several studies, progression of sepsis required the TLR adaptor molecule MyD88 and antibody mediated inhibition of TLR4/MD2 prevented lethality induced by CASP [31;32]. IL-12 and inducible nitric oxide synthase (iNOS) offer some antibacterial and immunoprotective effects since mice genetically deficient of IL-12 and iNOS are more susceptible to CASP-induced sepsis [33]. Additionally, exogenous addition of complement factor C3 [34] and activated protein C [35] offered cytoprotective and anti-inflammatory effects against CASP-induced lethality.
The CASP model of polymicrobial sepsis and peritonitis appears to have substantial utility, however our understanding of signalling pathways and the pathobiology which results in disease in this model are lacking. Animals subjected to CASP appear to mount a rapid, stronger immune response than animals subjected to CLP but further experimentation is required to determine the efficacy of using CASP as a sepsis model. Undoubtedly, the use of CASP will increase in prevalence as the pathophysiology and underlying mechanisms of disease which produce the septic conditions are uncovered.
Notable differences in the host response to CLP versus CASP-induced sepsis have been documented. The progression of sepsis in the CLP model involves TNFα-mediated activation of the immune response while the CASP model is less dependent on the initial immune response elicited by TNFα and more so dependent on TLR activation and signalling [12;25;29]. One study comparing the two distinct models found that bacterial counts of peritoneal lavage, liver, lung, and serum levels of TNFα, IL-1β, and IL-10 increase steadily over a 24 hour period in the CASP model and were significantly higher than that of mice subjected to CLP [24]. In this study, the authors observed continuously low bacterial counts and cytokine levels at all time-points as well as abscess formation around the cecum in mice subjected CLP. In light of these observations, it has been suggested that CASP is a true model of peritonitis with early SIRS, while CLP more closely mimics abscess formation [24;25]. Alternatively, others interpret the rapid elevations in systemic cytokines, bacterial counts and strong immune response following CASP to be comparable to that observed in endotoxemia models and the inflammatory reactions characterized by protracted cytokine profiles following CLP to more closely reflect clinical sepsis [12].
Discrepancies between the host response to CLP versus CASP have been observed in studies using genetically-modified animals, although consensus on the interpretation of these results is lacking. While TNFα-deficient mice were protected in a CLP model, TNFRp55/TNFR1-deficient mice did not appear to have an altered resistance to CASP-induced sepsis [26;36]. However, the differences observed may be due to the incomplete abolishment of TNFα activity as these mice may be deficient in only one of two TNF receptors, abolishing the cytotoxic TNFR1 and leaving the protective TNFR2 intact [37]. Additionally, IFNγ exhibited protective effects in a CASP model and not in CLP-induced sepsis [26;36]. Abolishing cytokine IL12p40 rendered the host more susceptible to CASP-induced sepsis while the same deficiency was found to either have no significant effect in some CLP studies [36] or increase susceptibility to sepsis in others CLP studies [38]. The discrepancies in experimental outcomes of studies using CLP and CASP may be results of differences in the host response between these sepsis models, and an incomplete understanding of the pathophysiology of each model. Careful consideration should be taken to choose an appropriate model to address the primary research question to be investigated.
Earlier models of endotoxemia and bacterial inoculum fall short in modelling the complex changes which occur in clinical sepsis. Many cardiovascular, respiratory, metabolic, hormonal, inflammatory, innate and adaptive immune changes associated with the spectrum of septic conditions cannot be sufficiently reproduced by a single injection of endotoxin or bacteria [12]. Injection of isolated microorganisms fails to mimic the host response to the diversity of causative agents in clinical sepsis. A specific instance is the injection of an endotoxin like lipopolysaccharide (LPS, a component of the cell wall of gram-negative bacteria) [4]. LPS endotoxemia is dependent on TLR4 signalling and represents one specific aspect of the immune response, not the complex interactions of multiple signalling pathways during the progression of sepsis. Endotoxin injection and bacterial inoculum are followed by rapid elevations in cytokines which are much higher than what is observed in human sepsis [4]. These models are more reflective of endotoxic shock rather than sepsis due to the overload of endotoxins in murine animals which exhibit a much higher endotoxin resistance than humans, further decreasing the clinical relevance of these studies [12]. Moreover, these increases are transient, occurring in a short time span, and fail to reflect the complex physiological response in clinical sepsis.
Although surgical polymicrobial sepsis models show a greater clinical relevance than earlier models of endotoxemia and bacterial or fecal injections, both CLP and CASP are not without limitations. While the severity of experimental sepsis can be controlled by modifying certain elements of a CLP protocol (e.g. length of ligated cecum) these factors may also decrease the consistency between animals and reproducibility of the study, as the amount of cecum ligated may vary between subjects [3]. More contributing factors to inter-animal variability include differences in the amount of fecal content in the cecum at the time of surgery, the size of cecum of each animal, and bacterial flora in different animals. Moreover, conflicting findings over the effect of the number of punctures on disease severity further highlight differences that may result due to surgical manipulation at the hands of different experiments [24]. Another limitation of CLP and CASP……..
Another limitation of CLP and CASP is the inter-study variations due to differences in protocols used between investigators. The number of cecal punctures and sizes of needles used to perforate the bowels vary between studies using the CLP model. Likewise, differences in the diameter of catheter used, location of stent insertion and suturing in the CASP model also influence disease severity [24;25]. While some of these limitations will inevitably affect the consistency and reproducibility of sepsis studies in animals, standard protocols can be enforced to reduce potential discrepancies.
Small size, shorter reproductive cycles as well as less housing and maintenance costs are some advantages of utilizing small animals for scientific research. However certain physiological features of small animals vary considerably from its human counterparts [39]. In addition, serial tissue and blood samples cannot be extracted from small animals, increasing the number of animals required to study whether an affect exists. Large animals, on the other hand not only allow serial sampling but also have very similar immunological and physiological functions to humans, rendering them better subjects to model clinical sepsis and drug testing. The Food and Drug Association (FDA) also recognizes the value of large animals requiring all new drug applications to include data from at least one non-rodent animal. In this section the rabbit, canine, porcine, ovine and non human primate models of sepsis and their limitations will be discussed.
A rabbit model of pneumococcal sepsis was developed in 1970. It encompasses several aspects of clinical sepsis including increased cardiac output and body temperature [40;41]. This model involves the use of
The canine models of endotoxemia and bacteremia have been used extensively to study cardiovascular function during sepsis. Dogs subjected to septic shock, show responses that parallel human sepsis. For example, there is a severe but reversible decrease in systolic ventricular function [44], a 32-108% increase in cardiac output, decrease in mean arterial pressure, and leukocytosis as well as increase in plasma epinephrine and norepinephrine levels during septic shock [45]. Cytokine profiles in the canine model of endotoxemia also mirror those reported in human sepsis patients with a 62 fold increase in IL-6 mRNA in peripheral blood mononuclear cells (PBMC) and 4.5 fold increase in TNFα within the first hours compared to controls [46].
Despite the analogous physiological responses, there are several limitations of this model pertaining to clinical relevance. Canines have an adrenergic sensitive sphincter around the hepatic vein which constricts during sepsis increasing intestinal venous pressure and damaging the mucosal barrier, increasing its relevance to a gut injury model [47;48]. Another important limitation is the resistance of canines to endotoxins and the requirement of high LPS dosage to induce sepsis [39;49]. This requirement for increased endotoxemia results in a severe hypodynamic response in canines [50] which does not mimic hyperdynamic human sepsis.
Pigs are popular animals for since they are readily available and relatively easy to handle. Continuous infusion of live bacteria, endotoxins as well as CLP are some of the methods used to induce sepsis in pigs. Porcine models of sepsis have been used extensively to investigate therapeutic agents focusing on improving renal, hepatic, intestinal and cardiovascular function [51-53]. For instance, using a porcine model of LPS induced shock, Cohen et al. observed that increasing levels of nitric oxide (NO) improves renal blood flow and glomerular filtration rate [51]. In addition, the porcine model of fecal peritonitis has been previously used to investigate the impact of adding low dose arginine vasopressin (AVP) to norepinephrine infusion for improving organ function [52]. In the AVP treatment group, renal function was significantly improved, and significantly less hepatic apoptosis compared to the group treated with norepinephrine alone, suggesting that the addition of AVP to norepinephrine improves renal function [52].
The porcine model of fecal peritonitis has also been used to study acute lung injury associated with sepsis. Septic pigs have decreased left ventricular function, respiratory dysfunction as well as hemorrhage, pulmonary congestion associated with neutrophil infiltration characteristic of acute lung injury [53]. This model has been further used to study effects of certain therapeutic agents such as N-acetylamrinone and L-arginine [54;55].
Due to the close similarity between pig and human anatomies, these animals have also been utilized to develop techniques commonly performed on sepsis patients such as laparoscopy [56]. Porcine models of neonatal sepsis also closely mimic the clinical course of neonatal sepsis with a significant decrease in systemic blood pressure [71 ± 3 mmHg in sepsis and 64 ± 3 mmHg in control at 3 h) and increases in serum levels of endotoxins, TNFα, and IL-6 [57]. Anatomical and physiological similarities between porcine and human anatomy have allowed for the successful testing of techniques and study of therapeutic agents which translate to human sepsis.
In sheep, infusion of endotoxins [58], live bacteria as well as administration of fecal content into the abdominal cavity are some methods used to induce sepsis. The ovine model of sepsis has distinct similarities to human sepsis and several advantages over other animal models specifically regarding cardiopulmonary responses. One similarity between sheep and human sepsis is the biphasic cardiovascular profile. In endotoxemia models of ovine sepsis, two phases of cardiopulmonary function are observed [59]. Within the first hour, the animal is in a hypodynamic state with a low cardiac index, myocardial contractility, high mean pulmonary arterial pressure, and pulmonary vascular resistance [59]. The first phase is followed by a hyperdynamic state with significant increase in cardiac output [59].
The ovine is also a popular specie to study sepsis associated with lung injury. Daniel Traber is a pioneer in developing the ovine model of \'smoke injury\' which involves insufflating sheep with smoke from burning cotton cloth [60] and the \'ovine burn model\' which involves third degree burns and smoke injury [61]. Smoke inhalation is induced using a modified bee smoker filled with 40 g of burning cotton cloth attached to a tracheostomy tube through a modified endotracheal tube [61]. Based on similar principles, the ovine model of \'smoke inhalation and septic shock\' was developed to study sepsis associated with pneumonia that develops due to acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) after smoke inhalation [62]. In this model, the aforementioned technique of \'smoke inhalation’ is used to induce lung injury [63], followed by instilling bacteria such as
Using the ‘ovine smoke inhalation and septic shock\' model several therapeutic agents have been investigated for sepsis management. Necrosis and apoptosis in this model were improved by administration of WW-85, a peroxynitrite decomposition catalyst. Other notable changes include improved gas exchange, decreased levels of myeloperoxidase (MPO) and lung 3-nitrotyrosine [65]. There was also an overall improvement in pulmonary function suggesting that blocking nitric oxide-peroxynitrite pathway may ameliorate some effects of septic shock [65].The ovine model of sepsis and septic shock has made important contributions in the areas of cardiovascular [66] and pulmonary research as well as in determining the efficacy of fluids [58] and therapeutic agents [67], such as anticoagulants [68] and recombinant human protein C [69] to improve sepsis outcomes.
Due to genetic similarities to humans, non-human primates are an ideal model for testing species-specific therapies [70]. The method of inducing sepsis and septic shock in baboons utilizes an intravenous infusion of live
Large animal models have provided tremendous insight into the pathophysiology of sepsis, however there are certain limitations to the use of these animals. Due to their large size, they are also more difficult to handle, house, and anaesthetize and in the case of primates, pose risks of cross transmissible diseases [47].
There have been several therapeutic agents with positive outcomes in animal models however majority failed to show efficacy in clinical settings. This disconnect is partially due to discrepancies in translating findings from animal models to clinical sepsis. Defined human target populations and established severities of sepsis would allow for more realistic and applicable animal modelling.
Clinical studies report that individuals with co-morbidities such as diabetes are at a higher risk of mortality and morbidity from infectious diseases [74-77]. It has also been estimated that greater than 50% of septic patients have at least one additional co-morbid condition [78]. Therefore, incorporating co-morbid conditions into animal models is one method by which modeling clinical sepsis in animal models could be improved.
Several animal studies and clinical trials have demonstrated that diabetes increases the risk of infections and mortality [74-77]. A retrospective cohort study from Ontario, Canada by Shah et al. reported that almost half of the diabetic patients in their study (n=513,749] had a minimum of one case of infectious disease for which they were either hospitalized or received a physician claim [74]. It was also found that bacterial infections and infections in general were also more commonly found in diabetics [74]. From animal models of type II diabetes it is known that sepsis induced inflammation is more severe in diabetic animals [79]. Septic diabetic animals also have increased bacteria load [80;81], altered levels of cytokine expression [81;82] and dysfunctional immune cells such as PMNs contributing to poor outcomes.
Studies in animal models of obesity parallel the findings in diabetic animals. Strandberg et al. reported that mice fed a high fat diet (HFD) with saturated fats, had higher mortality rates when challenged with
The urogenital tract is the focus of infection in approximately 25% of all sepsis cases [84]. Urinary tract infection (UTI) associated sepsis is predominantly found in elderly individuals, diabetics and immuno-suppressed patients [85]. Several animal models have been developed to study different aspects of these co-morbid conditions. Harberg et al. developed a model of
Several studies have identified the implications of genetic variance, for immune responses to sepsis. Even while comparing across different strains of septic mice, mortality rates, liver MPO activity, metallothione mRNA, leptin as well as IL-10 levels are significantly higher in C57BL/6J compared to A/J mice [88]. Thus, genetic differences in mice and possibly humans are associated with differences in the inflammatory processes initiated in response to infection that ultimately affects sepsis outcomes [88].
Transgenic animal models have also been utilized extensively in sepsis research [89]. Transgenic apolipoprotein (ApoE) polymorphs, ApoE3TR and ApoE4TR, generated by replacing mouse ApoE allele with its human counterpart coding sequences were used to investigate if ApoE genotype affects sepsis outcomes [89]. ApoE is a ligand for low density lipoprotein (LDL) receptor, responsible for clearance of VLDLs and chylomicrons residues. ApoE plays key role in lipid metabolism [90] and mediates removal of inflammatory apoptotic substances [91]. Mice with the human APOE4 allele have greater inflammatory response (with two to four fold increase in synthesis of cytokine) as well increased time to mortality [89].
ApoE knockout mice (apoE -/-) generated by gene targeting are more susceptible to endotoxemia and K.
Other strains of transgenic mice have also been utilized for sepsis research such as mice with inactivated beta2 integrin CD11b. These transgenic mice are partially protected against micro-vessel permeability and edema formation, suggesting key role of Cd11b in lung PMN sequestration and vascular injury during the early phase of gram-negative sepsis [94]. Lectin-like oxidized low density lipoprotein receptor 1 knockout (LOX-1-/-) mice have been used to investigate the role of LOX-1 in sepsis induced mortality. Wu et al. reported that LOX-1-/- mice with CLP-induced sepsis had decreased systemic inflammation, neutrophil migration to sites of infection, levels of pro-inflammatory cytokines, and lung edema as well as increased bacterial clearance implicating the key role of LOX-1 in systemic inflammation [95]. Important findings have also been discovered in toll like receptor knockout mice (TLR-/-). Toll like receptors (TLRs) recognize different components of bacteria cell wall and mediate immune responses to infection. TLR-4 for instance interacts with LPS, activating the expression of target genes. TLR-/- knockout mice have been utilized to study dendrititc cell maturation and cytokine production [96], infections of the urogenital organs [97] and defense against infections such as murine tuberculosis [98] among several others aspects of adaptive and immune responses.
Transgenic mice such as ob/ob have also been used to examine the effects of leptin deficiency on sepsis outcomes [99]. Leptin deficiency, in ob/ob mice, is associated with greater organ dysfunction and increased mortality rates during sepsis [99]. Leptin signaling appears to improve survival and may be required for immune responses, implicating therapeutic potential for leptin analogues for sepsis. Thus transgenic mice have made important contributions to Improve our understanding of sepsis at physiological, immunological and cellular levels.
An emerging area of sepsis research involves the formation of neutrophil extracellular traps. In 2004, Brinkmann et al. characterized a novel mechanism of innate immunity exhibited by neutrophils. Upon stimulation by endotoxins or pro-inflammatory mediators, activated neutrophils released chromatin material composed of a DNA backbone and antimicrobial granular proteins in the form of neutrophil extracellular traps (NETs) [100]. NETs ensnare circulating microorganisms, preventing further dissemination of pathogens in the vasculature while providing a scaffold for neutrophil granular proteins. This creates a high, local concentration of proteins with antimicrobial properties [100] and allows for microbicidal synergy, concentrating their ability to disarm and kill microorganisms [101].
The formation of NETs, recently coined NETosis [100;102] is an active process. Cleavage of histones by neutrophil elastase is sufficient to cause decondensation of nuclear material [103] which can be observed before disintegration of nuclear and granule membranes. Chromatin material mixes with nuclear and granular proteins with potent antimicrobial properties and is released extracellularly [102;104] forming stretches of DNA and globular proteins with diameters of 15-17 nm and 25 nm respectively which can aggregate to form thicker threads around 50 nm in diameter [100]. The process of NETosis is distinct from that of necrosis (the plasma membrane remains intact while nuclear and cytoplasmic granular components mix) and that of apoptosis (no phosphatidylserine exposure signaling phagocytosis, or nucleosomal cleavage) [102;105].
NETosis can be experimentally induced by exposure to endotoxins (e.g. LPS), gram-negative and gram-positive bacteria, fungi, pro-inflammatory mediators (e.g. IL-8, phorbol myristate acetate or PMA a protein kinase activator), and activated platelets [103;106-111]. NET formation in experimental conditions results in the release of proteins which degrade virulence factors expressed on the pathogen surface and create a toxic environment for invading microorganisms [102;107;112-115]. NET-associated azurophilic granule proteins which have antimicrobial or immunomodulating effects include histones with specific post-translational modifications (and histone cleavage products like buforin), cathepsin G, elastase, MPO, pentraxin, gelatinase (matrix metalloproteinase-9], catalase, lactoferrin, peptidoglycan recognition proteins (PGRPs), and bactericidal permeability-increasing protein (BPI) [100;107;116]. NETs have also been shown to kill infectious organisms and impair pathogenic invasion of gram-negative bacteria (e.g.
In 2007, a landmark paper published by the Kubes group characterized the mechanism by which platelet-neutrophil interactions contribute to pathogenesis of severe sepsis. It was observed that LPS-induced endotoxemia in mice as well as plasma from severe sepsis patients were able to trigger NET formation but at a cost to the host. Upon detection of TLR4 ligands (e.g. LPS), platelets bind to neutrophils adhered to the endothelium [118]. Within minutes, TLR4-dependent platelet-neutrophil interactions resulted in the robust release of granules and DNA in the form of NETs the integrity of which was maintained under flow conditions reflective of physiologic shear forces in the microvasculature [0.5 dyne/cm2]. NET-mediated bacterial clearance through trapping and ensnaring of bacteria was primarily observed in liver sinusoids and lung capillaries, areas with the greatest capacity for immobilizing bacteria due to the decreased lumen size of vessels [118]. However, formation of NETs is not without a cost as it was also found to cause cellular and tissue damage to the host.
NETs induce pathogenic effects via multiple mechanisms. Platelet-mediated NET formation trapped platelets, leukocytes, and red blood cells causing microvascular plugging and areas of ischemia in downstream tissues [118;119]. Ischemic conditions increase the production of IL-8 [120] and reactive oxygen species [121] both of which can induce further NET formation [122]. Several lines of evidence also suggest that NETs exert pro-coagulant effects in sepsis and other conditions of deregulated coagulation and inflammation. In studies of thrombosis, NETs were found to interact closely with fibrin and platelets via von Willebrand factor, fibronectin, and fibrinogen, effectively stabilizing platelet-rich clots [119;123]. NETs perfused with blood or platelet-rich plasma stimulated platelet aggregation and promoted thrombus formation [124]. Another
NETs and NET-associated proteins have also been found to exert pro-inflammatory effects. MPO released during NET formation binds the negatively-charged, proteoglycan-rich endothelium, inducing endothelial cell damage and vascular permeability [130;131]. Incubation of NETs with human platelets and THP-1 cells resulted in the release of pro-inflammatory cytokines including IL-1β, IL-8, and TNFα [132].
The pathogenic nature of NETs is also supported by studies showing improved outcome associated with NET destruction via DNase administration [133]. Furthermore, DNaseI-deficient mice develop lupus-like symptoms induced by NET formation, indicating that NET removal is likely a crucial process in the proper immune response [123]. The formation of NETs and release of NET-associated proteins (e.g. histones) potentiate the pro-inflammatory and pro-coagulant response, both of which contribute to end organ morphology in sepsis. These changes include neutrophil adherence to microvascular endothelium, inflammatory infiltration, vacuolization of epithelial and endothelial cells, fibrin deposition, microvascular ruptures with intra-alveolar hemorrhaging, and the formation of platelet-rich micro- and macrovascular thrombi [118;127;129;134;135]. While clinical isolates have been shown to have the ability to generate NETs
Despite the success of many therapeutic agents in improving outcome in preclinical studies of sepsis, many have failed to demonstrate efficacy in the clinical setting. To illustrate, studies investigating the efficacy of anti-TNF therapies [5;5;136;137] and activated protein C (APC) were promising in animal studies, but this success failed to translate to clinical outcomes [35;73;138]. Neutralization of TNF was beneficial in some models of endotoxemia challenge with viable gram-negative bacteria (e.g. group B streptococci) [139-145] but in other preclinical studies where microorganisms such as Candida, S. pneumoniae, Listeria, or mycobacteria were used, neutralizing TNF exacerbated outcome in animals subjected to the pathogenic challenge [36;70;136;137;146-149]. In complex models of sepsis, no consistent harm or benefit was validated and some clinical studies found an increased mortality associated with anti-TNF therapy [150;151]. Similar to anti-TNF therapy, some preclinical studies demonstrated the protective or therapeutic effect of APC [35;73;138] but these results failed to translate in some clinical studies [72]. The therapeutic efficacy of anti-TNF therapy and APC in preclinical studies is largely influenced by the animal model used. Thus the appropriate translation of findings and inferences from preclinical studies to clinical outcomes in sepsis remains a heavily debated area.
Limitations in both the use of animal models and design of experimental studies contribute to the poor translation of preclinical animal studies to clinical sepsis. Some limitations are inherent to the use of animals to model any clinical condition. It is commonly the case that young animals of a specific gender, species, genetic background, and nutritional status housed in a pathogen-free, sterile facility unexposed to the natural environment are used [5]. Many of these elements are tightly controlled to maintain consistency at the expense of clinical relevance. However, an attempt at balancing both consistency and clinical relevance can be made if the investigator designs treatment groups for different animals (e.g. separate treatment groups for females and males, groups for young and aged mice, etc.) given the heterogeneity of the sepsis patient population.
Other limitations may be appropriately addressed by establishing clear research questions and implementing an experimental protocol which would adequately investigate these questions. For instance, if the objective of an animal study is to test the clinical applicability of a therapeutic agent, it would be more clinically relevant if therapies currently used for the management of sepsis including the adequate administration of resuscitation fluids, antibiotics, and supportive therapies [14] were incorporated into the experimental protocol of the animal study. Additionally, the experimental protocol of animal studies can be modified to include clinically relevant management procedures such as constant monitoring and assessment for hemodynamic parameters, tissue perfusion, or dehydration as would occur in clinical sepsis. Other factors which may contribute to the gap between findings in experimental and clinical studies include the time at which therapeutic agents are administered, the lack of staging of sepsis to reflect disease progression at different severities on the sepsis continuum or different patient populations, and the risk of experimenter bias in animal studies [5;12]
It is commonly the case that murine studies of sepsis utilize 8 week-old mice, the physiological equivalent of a young adulthood in humans. However, the sepsis patient population consists largely of patients over 60 years of age which is not adequately represented in sepsis literature using animal models [152]. There are limitations to examining the true pathophysiology and clinical treatment of a condition which occurs most commonly in the aging population when findings are extrapolated from preclinical studies using young, healthy mice exclusively. For example, aging is associated with increased apoptosis of rapidly dividing epithelial cells of the gut and spleen in animal sepsis [153]. This may contribute to the increased mortality in aged septic mice, a factor which would not accounted for when using young animals to model sepsis occurring in an aging population. The effect of age in sepsis in a clinical setting can be appreciated in the PROWESS trials. Substantial difference in absolute risk reduction of mortality were found to be associated with age in clinical sepsis [154]. Age should be considered in animal studies of sepsis to increase the clinical relevance to human sepsis.
In addition to limitations produced by using young animals exclusively, male animals are almost exclusively chosen in intra-abdominal sepsis studies which poorly reflects the incidence of clinical sepsis in both genders around 40% of which is female [155]. Male mice are often chosen over females to avoid confounding effects and variables posed by varied expression of biomarkers and circulating cells associated with different phases of the estrous cycle. For instance, steroid hormones are implicated in the expression of adhesion molecules resulting in different peripheral blood leukocyte concentrations and an altered coagulant response [156;157]. Neutrophil concentrations and response to stimulation also vary considerably during different phases of the estrous cycle [158]. For consistency, male mice are often chosen of sepsis studies, despite the similar occurrence of sepsis in males and females however, an investigator should consider, if feasible, treatments with one group for each gender of animal used.
The importance of early goal-directed treatment including adequate fluid resuscitation and treatment with antibiotics has been thoroughly demonstrated in severe sepsis and septic shock [159]. Correction of hemodynamic abnormalities and hypovolemia associated with sepsis is integral to reducing mortality rates and improving sepsis outcomes. Hypovolemia compromises tissue oxygenation due to inadequate blood flow within the microvasculature and is the prime cause of organ dysfunction and failure [159]. In order to differentiate pathology due to sepsis from pathology resulting solely from circulatory decline and lack of hemodynamic support, the administration of balanced fluids is crucial. This is supported by findings showing significant differences between the hemodynamic profiles of under-resuscitated animals versus those with adequate supportive fluids, and aggressive fluid resuscitation was required to replicate hemodynamic profiles observed in patients with severe sepsis [47]. In canines with septic shock, animals that received combination therapy of antibiotics and cardiovascular support (via fluid resuscitation and dopamine) had a 43% improvement in survival rates compared to septic animals treated with either therapy alone [160;160]. Furthermore, experiments elucidating the effects of various fluid regimes on resuscitation in sepsis have demonstrated that lactated Ringer’s crystalloid solution but not saline-based solutions reduced sepsis-induced leukocyte recruitment in the liver of mice subjected to CLP [161]. Based on these studies, an appropriate fluid regime which would account for surgical losses and provide adequate hemodynamic support to maintain circulatory and cardiovascular function, as would occur in the management of clinical sepsis, should be considered in preclinical studies of sepsis [12].
The human septic population is diverse and highly heterogeneous. Clinical cases of sepsis are often much more complex than sepsis induced in animal studies. The diversity of infectious agents as well as sites of infection in clinical sepsis are not always reflected in animal studies, factors which should be considered in the translation of preclinical to clinical studies. For instance, clinical sepsis may result from trauma and subsequent fecal spillage into a sterile peritoneum or staphylococcal bacteremia in an elderly patient with congestive heart failure. Evidently, animal models like those inducing sepsis via injections of endotoxic bacterial components or even live bacteria fail to induce the range of conditions observed in a septic patient [5]. To decrease this gap, one may choose to consider introducing cormorbidities as previously described or other trauma or infectious injuries to animal sepsis models.
Animal | Species, genetic background, gender, age, nutritional status following insult, comorbidities incorporated in model |
Source of Infection | Single versus multiple organisms, local versus systemic challenge, addition of adjuvants, presence and extent of tissue damage from challenge |
Intervention | Dose and timing of intervention compared to septic insult (administered before, concurrent with, or following insult) |
Co-interventions | Fluid resuscitation, antibiotics, analgesics, source removal |
Experimental Design | Risk bias (blinding or randomization methods are used), assay methodology |
Markers of Outcome | Parameters used as markers of outcome (choice of biomarker and quantification methods, physiological response, inflammatory parameters, survival) |
Limitations to the Clinical Relevance of Preclinical Studies
Factors which limit the translation of preclinical studies to clinical outcomes include the following [5]: the animal used for experimental studies (species, genetic background, gender, age, nutritional status following insult, comorbidities incorporated in model), the source of infection (organism, local vs. systemic challenge, adjuvants, tissue damage from challenge), dose and timing of intervention (administered before, concurrent with, or following insult), co-interventions (fluid resuscitation, antibiotics, analgesics, source removal), experimental design (risk bias, blinding, randomization, assay methodology), and parameters used as markers of outcome (quantification and choice of biomarker, physiological response, inflammatory parameters, survival) which can significantly alter response to treatment. These potential limitations are summarized in Table 1 adapted from Marshall et al, 2005.
It is evident that experimental design greatly influences the findings of both experimental and clinical studies in sepsis. One method to increase the transparency, reproducibility, consistency, and efficacy of sepsis research is to increase standards for reporting of animal sepsis studies. The Consolidated Standards of Reporting Trials (CONSORT 1996, 2001, and 2010] statement is a rigorous and highly standardized approach to conducting clinical research which addresses variability of results due to the study design and reporting of clinical trials [162-164]. In an attempt to limit the variability in both the use of animal studies and reporting of results, Marshall et al. propose that a similar checklist should be required for preclinical studies [5]. The checklist includes the following variables which should be explicitly described and recorded in detailed manner to ease pooling of results from various preclinical studies, define a framework from which to understand divergent results, improve the consistency of reporting of results, and enhance the reliability of reported results. The standards for reporting animal research in bioscience were raised further in 2010 with the publication of the ARRIVE guidelines, Animals in Research: Reporting
In the design and reporting of a preclinical sepsis study, a basic but crucial aspect that one should consider is the very definition of sepsis, the conditions and presentation of which differ from one animal model to another. Clinical sepsis is defined by meeting specific criteria which involve inflammatory, hemodynamic, organ dysfunction, and tissue perfusion variables. Sepsis is no longer defined solely by changes in physiological parameters (e.g. body temperature, heart rate, respiratory rate, and white blood cell count) which are common to many other conditions. Likewise, defining sepsis based on levels of several biomarkers of inflammation, for instance fails to capture the complexity of cardiovascular, hormonal, metabolic, innate and adaptive immune changes which occur in this heterogeneous condition. Although it would be helpful to document changes in biomarkers and physiological parameters (e.g. heart rate, etc.), current preclinical studies are limited as the criteria which define sepsis (and the severity) in various animals have not been clearly elucidated. There is currently no consensus over the physiological parameters, levels of biomarkers, or variables of end organ health which would indicate sepsis in an animal although these parameters are much more clearly outlined by clinical sepsis definition guidelines [1]. This limitation will need to be addressed in future preclinical sepsis studies. Other parameters which may be informative of the septic condition in animals include appearance (perfusion of mucous membranes), urine production, and bacterial cultures from the blood, peritoneal fluid, and local site of infection, which may be considered in the design of animal studies. Successful animal studies should establish a clear definition of the conditions which would indicate a septic animal, explicitly defining the severity of disease induced as part of the study protocol [167].
Sepsis Definition | Clear sepsis definition as indicators of sepsis, severe sepsis, or septic shock in the animal used [166] |
Animal | Species, strain, genetic background, gender, age, weight, handling, housing, feeding conditions |
Experimental Design | Method of sepsis induction (details which indicate severity), intervention (timing and dose), experimental methodology, controls included, randomization methods, blinding of experimenter |
Analytic Plan | Primary and secondary endpoints, power calculations for sample size determination, intention-to-treat analysis, criteria for animals excluded from studies (substitute endpoints) |
Co-Interventions | Resuscitation fluids, antibiotics, source removal, feeding |
Results | Flowchart of included and excluded animals, establish mortality rates in studies conducted |
Checklist for the Reporting of Animal Studies
In the interests of reproducibility and reliability, the method of sepsis induction, intervention (timing and dose), experimental protocol, inclusion of proper controls, randomization methods, and methods of experimenter blinding should be documented in detail. Although experimental methodologies are commonly recorded, there is a significant lack of attention to reduce experimenter bias by randomization and blinding, despite the confounding effects recently addressed in a systematic review of literature of studies using animal sepsis models. It was observed that only 2% of systematic reviews and meta-analyses appraised the risk of bias or clinical relevance of the underlying animal research despite a significant proportion of the literature extrapolating results from animal studies to clinical sepsis [168]. In one review of studies using animal models, more than 80% of research papers surveyed did not report the use of methods to minimize the risk of bias by the investigator [165;169]. To improve the translation of inferences and findings from experimental sepsis to clinical sepsis in future studies, one should consider reducing the effects of experimenter bias by randomizing animals in the treatment groups of the study and blinding the experimenter to the intervention provided given that implementation of these elements would not compromise the protocol or results of the study [5;166;167].
In the absence of randomization, the results of clinical and animal studies may be unintentionally biased by factors which may influence study outcomes. Nonrandomized clinical studies have frequently shown a larger treatment effect than randomized control trials, an effect which should be studied and addressed in animal studies [168;170-173]. Randomization involves taking proactive measures in assigning subjects to a treatment or control group in studies testing the efficacy of a potential treatment drug versus a placebo control or potential treatment drug versus a pre-existing treatment [167;173]. The systematic randomization of subjects or animals minimizes allocation or selection bias by eliminating influences from unknown or known prognostic variables (like the hemodynamics in septic and nonseptic animals) which may influence the response to treatment, mortality, or outcome, unduly producing biased results in a study [166;170-172]. Randomization also ensures that protocols and procedures are conducted consistently and systematically in treatment and control groups. Elements of animal studies where randomization may offer an efficacious benefit include the procedure to induce sepsis (preventing unintentional bias by variations in sepsis severity), treatment (consistent timing, route, and method of administering treatments or controls), monitoring of subjects following sepsis induction and treatment, and resuscitation procedures of the study groups [169;173].
Although various methods of randomization have been established and recommended for different clinical trials, consensus and literature on randomization in pre-clinical and experimental studies are lacking. Simple randomization methods such as the "repeated fair coin-tossing" have been recommended for large clinical studies (n > 200], but are inadequate for studies with small samples sizes like experimental animal studies [167;173]. Restricted randomization methods like permutated-block randomization where allocation ratios are used to specify the number of subjects in each group (or ratio of subjects in one treatment group to another) [170-172] successfully address this issue, but restricted randomization methods have not been validated in animal studies. Given that the risk of allocation and selection bias have been shown to unduly influence the results of clinical studies and that randomization and blinding have successfully minimized these risks, it is crucial that the same level of criticism and caution be applied in the experimental design and conducting of experimental studies by addressing these risks [167;170-173].
In addition to randomization, blinding of investigators also minimizes the risk of bias at many levels of the preclinical study. Blinding decreases differential treatment of animals in the study groups (more intensive monitoring or closer supervision of one treatment group over another) to unintentional biases in analyzing data and adjudicating outcomes [5;166]. For instance, markers of disease severity or mortality outcome such as organ histology may be interpreted in a biased manner in an untreated control group versus the treated group if the investigator is not blinded to the treatment received [169]. Likewise, marginal findings may be analyzed in a biased manner when interpretation of the outcome marker is subjective. In studies where substitute endpoints are used (such is the case when mortality is not used as endpoints for ethical reasons), blinding reduces the chance of experimenters prolonging the time to endpoint (in an attempt to establish a therapeutic effect when one does not exist) or decreasing the time to a substitute endpoint or even excluding animals (if unexpected results are observed in either treatment or control groups) [165;167;170;172]. Blinding of the investigator to the treatment or control substance received minimizes the risk of experimenter bias.
The analytic plan should be described in detail such that the procedure is reproducible by others and variations in models used by other investigators can be appreciated (e.g. size and number of enterotomies and location of ligature in a CLP model) when comparing various preclinical studies [5]. Primary and secondary endpoints of the study should be clearly outlined, whether they are to determine the efficacy of a potential therapy, improve prognosis, or diagnosis of sepsis. If the primary objective is to investigate a potential therapy, endpoints and markers of outcome of the animal study should also be clinically relevant [166;173;174]. The measurement of outcome should also appropriately reflect markers relevant to the potential therapy being tested
In studies elucidating the therapeutic potential of a compound, power calculations can be used to reliably determine the sample size required to show that an effect is associated with treatment as well as exclude an effect when none exists [5]. The magnitude of a treatment effect is most commonly expressed as the relative risk reduction (RRR, RRR = [1-(X/Y] x 100 where Y is the proportion of animals that expired in the treatment group and X is the proportion of animals that expired in the control group [175]. Given that mortality (and therefore the power of the study) will be altered should any subjects be excluded, the exclusion of subjects should only occur if subjects meet pre-established criteria.
As in clinical studies, conditions under which subjects will be excluded should be clearly outlined before the animal study [169]. The study should account for all animals, including those that do not complete the experimental protocol and observations and reasons for excluded animals should be appropriately documented. This is important to determine whether prognosis independent of the treatment may in actuality be related to the exclusion of certain subjects [5;174]. Data should also be recorded and kept for subjects excluded from the study for any post-hoc analyses, standards which are expected of clinical studies.
Many animal ethics and welfare committees are preventing the use of mortality as an endpoint and encouraging the use of surrogate markers of death. However, investigators and funding agencies are critical of such studies which do not use changes in mortality as a measure of therapeutic efficacy. There is currently no consensus nor scientific validation of clinically relevant, reliable substitute markers of death in septic animals [5;174]. Moreover, there is concern over the use of analgesics and anaesthetics which have been shown to interfere with components in the natural progression of sepsis and even exert protective, anti-inflammatory effects. For example,
Given the potential limitations of preclinical studies when designing preclinical studies with potential clinical applications, one should take a critical approach when validating the therapeutic efficacy of a potential drug in preclinical studies. Recommendations for conducting preclinical studies with clinical applications will be provided with the intent to address the concerns raised previously. Literature by Marshall et al. suggest that early proof of concept studies should 1] delineate a potential pathological role for the target in question by quantifying its increases in a simple acute model like endotoxemia or bacteremia (LPS or E. coli challenge) and 2] demonstrate that attenuating such target may improve outcome or offer some therapeutic benefit by a measurable decrease in harmful effects of the initial challenge or insult [5;165;174]. If a proof of concept can be established, time course studies should be conducted to determine if the potential treatment offers therapeutic effects if administered before, during the progression of, or after the insult in both simple acute and complex models (e.g. LPS challenge and CLP). Experimental methodology and design of further studies should consider the results of studies on targets with similar biological or pathophysiological effects. Any adverse effects of the intervention, like impairment of the host’s natural anti-microbial immune function or further dysregulation of the coagulant state should be determined in a high-risk model using live organisms as the challenges. [5;174] Moreover, the systemic physiological and biological responses to the therapeutic intervention (e.g. cardiac output, oxygenation, glomerular filtration) should be determined in a large animal model, given the limitations of small rodent models of sepsis. Interventions used in the management of clinical sepsis such as fluid resuscitation, ventilatory support, and hemodynamic monitoring, can be incorporated into the study design of preclinical trials using large animal models [5].
Goals for the future use of experimental sepsis models ought to focus on improving the relevance, translation, and applicability of results and inferences from animal studies to what is observed clinically. This requires consideration of the limitations of current sepsis models described such as considering age, gender, genetic background, nutritional status, and the environment of the housing facility, incorporating cormorbidities and supportive therapies which may be clinically relevant, and finally addressing the risk of bias by randomization and blinding methods in preclinical studies of sepsis.
Given the importance of the organizational and interpersonal benefits shown from people’s ability to gain trust, it is crucial to see what influences how trustworthy a person is deemed. Past research has identified predictors of trustworthiness that include personality traits and physical attributes. Trait agreeableness and honesty-humility show positive correlations with trustworthiness [1, 2, 3]. Guilt-proneness—how guilty a person thinks they would feel about doing something wrong—was found to predict trustworthiness even better than agreeableness and other Big Five personality traits (i.e., extraversion, conscientiousness, and openness), and this relationship was mediated by interpersonal responsibility [2]. Facial expressions were also found to have a significant relationship with perceived trustworthiness [4, 5]. Although these elements are important to understand, there are additional influential factors of trust that still need more research. In this chapter, we underscore a growing body of research [6] that reveals one essential yet understudied personal trait that reliably impacts trustworthiness: self-control. Below, we review and highlight the role self-control plays in garnering trustworthiness, identifying the range of related positive and negative outcomes and questions for future research to explore.
Trust and trustworthiness are critical factors in social dynamics. Trust (a.k.a. propensity to trust) is described as the amount of vulnerability a person allows themselves in a particular situation [7]. Broadly speaking, trust occurs between two or more people, groups, or entities (e.g., romantic partners, co-workers, organizations with shared interests, an athlete and their team, and a political party and their candidate) and is characterized by feelings of confidence that the trustee will meet expectations of the trustor, which are generally positive or non-negative [8]. Trust is especially relevant in situations with no certain or guaranteed outcomes, where the trustor allows themselves to be vulnerable to the possibility that their expectations will not be met [9].
Separately, trustworthiness encompasses the perception the trustor has of the trustee’s ability to meet their expectations. It is often developed from past experiences and can differ depending on the context of the expectations. Past research has identified both a 2 and 3-dimensional model to conceptualize the way we understand trustworthiness. The 2-dimensional model suggests that trustworthiness is formed by both affect-based and cognition-based trust, where affect based-trust describes the belief or perception that the trustee will act in a manner that preserves the relationship, and cognition-based trust entails the trustee behaving in a competent and dependable manner [10]. The other model proposes three components that form perceptions of trustworthiness—benevolence, integrity, and ability [7]. A person who demonstrates that they [1] are capable of meeting an expectation, [2] without defying their accepted principles, and [3] without exploiting or taking advantage of the trustor’s vulnerability, would be viewed as trustworthy. While distinct, these models appear to overlap in their theory, as noted by Ferrin [11], who suggested that perceived ability and integrity signal cognition-based trust, and perceived benevolence communicates affect-based trust. Research has since supported this belief, finding that, for cognition-based trust, ability and integrity were better predictors, while benevolence was the best predictor of affect-based trust [12]. These findings overall support the idea that cognitive and affect-based trust are distinct from each other.
Trust and trustworthiness provide benefits to a wide range of situations. Research in management and organizational psychology suggests these traits lead to more productive workplace outcomes. For instance, in management settings, increased trust and trustworthiness in co-workers promotes openness, cooperation, information sharing, the exchange of ideas, opportunities for the development of beliefs and attitudes, and the acceptance of shared ideas [13, 14]. Trust has also been positively associated with job performance and citizenship behavior, and negatively associated with counterproductive behavior [15, 16, 17]. One study [18], found that “trustworthy managers preside over more productive organizations and are better able to maintain and even increase organizational outcomes in agencies challenged by low levels of performance and perturbations in the external environment.” More trustworthy managers were also associated with greater procedural and interpersonal justice in the workplace [19], and perceptions of trustworthiness provided by coworkers have been identified as predictors of work performance, specifically through impressions of ability and integrity [20].
Trust and trustworthiness are related to beneficial outcomes in non-work relationships as well. Interpersonal trust increases the closeness, quality and communication within interpersonal relationships with intimate partners, siblings, and children and parents [21, 22, 23, 24, 25]. Rotter [25] found that those who are more trusting are less likely to participate in immoral behaviors such as lying, stealing or cheating, and have a decreased likelihood of being maladjusted or unhappy. Divergently, high trustors are more likely to respect the rights of others, give second chances, be desired as a friend more, and be more well liked [25]. Greater perceived trustworthiness was found to contribute to peer acceptance, school adjustment and performance [26], and was positively related to developing relations with peers and having more friendships [26, 27, 28]. Considering the many benefits that trust and trustworthiness can produce, it is advantageous and important to understand what predicts it, and some recent research has identified self-control as a signal of trust.
Self-control has been vastly studied through the decades, with over 2 million related search results on Google Scholar as a testament to the topic’s importance. By definition, self-control is the regulation of behaviors and thoughts to pursue a more distant and abstract goal or motive when a directly conflicting opportunity to satisfy an immediate and concrete motive or goal is present [29, 30]. In other words, self-control is demonstrated in your decision to forgo the tasty treat that is currently available to you, in order to remain committed to your diet and long-term goals of a healthy lifestyle. It is no surprise that this characteristic, and its related outcomes, have maintained the interest of psychologists for so long.
The outcomes related to self-control are as important as the trait itself. Research has identified that trait self-control is positively linked to better physical health and performance in school and work [30, 31, 32], along with greater attainment and subjective well-being [33, 34]. It has also been shown that those with greater self-control show more empathy, perspective taking, less deception, and report better behaviors in romantic relationships [21, 35, 36]. These positive associations with self-control have naturally led researchers to investigate how individuals can increase this beneficial trait. Past findings have recommended methods related to goal setting, monitoring, and implementing [37], and described various types of interventions (i.e., social skills development programs, cognitive coping strategies interventions, video tape training/role-playing interventions, immediate/delayed rewards clinical interventions, and relaxation training) that have helped increase self-control and reduce delinquency in children [38]. Practicing mindfulness and small acts of self-control, such as eating fewer sweets, has also led to improved performance on self-control tasks [39].
While having trait self-control and being seen as someone who demonstrates self-control are not necessarily the same, positive consequences have been identified based on mere perceptions of the trait. A person who is viewed as being self-controlled has better social relationships, with greater satisfaction and success [31, 40, 41]. Perceived self-control is also related to greater organizational outcomes like being viewed as more fair at work [42]. However, these perceptions have also been associated with some negative outcomes including assumptions that the work done by highly controlled individuals is less arduous and time-consuming, which can lead to the employee being overburdened with extra assignments [43]. High perceptions of self-control can also cause an individual to face negative consequences in social settings where their company may be less desired [44] or in academic settings where their peers may be less likely to offer them assistance [45].
While self-control is commonly considered an intrapersonal trait, our perceptions of other people’s self-control are important signals during interpersonal settings. The amount of self-control a person demonstrates significantly impacts other perceptions we have about them, which can ultimately influence our behaviors and attitudes towards the person.
As suggested by an increasing amount of recent empirical evidence [6, 21], we argue that perceptions of self-control function as a reliable signal for trustworthiness. Below, we summarize the varied emerging evidence and elaborate on how self-control induced trustworthiness manifests across different relationship contexts.
In
Notably, while there are many upsides of high self-control perceptions in organizational relationships, some downsides have been identified by more recent research as well. While high self-control individuals benefit by being trusted and desired more as partners in work-related settings [44], they are also relied on more, and have more expected of them by their workmates, compared to those perceived as lower in self-control [43]. Their associates also tend to think the work done by those with high self-control takes less effort and is easier [43]. High self-control people may then be asked or expected to complete more tasks because their associates trust them to meet the heightened expectations, while receiving less recognition compared to their co-workers who are lower in self-control. These beliefs can lead to high self-control individuals feeling overburdened and underappreciated, resulting in a decrease in relationship satisfaction [43].
Organizational relationships are not alone in their potential for negative consequences of high self-control perceptions. Research by Röseler [44] has found that, while those perceived as having greater self-control are preferred in settings of work, they are less preferred in social settings, such as parties, compared to people with lower levels of self-control. This may result from the belief that the high self-control person, who suppresses desires and forgoes immediate satisfaction in pursuit of long-term goals, will continue to meet that expectation as they have previously. If it is trusted that these expectations will be maintained, then the person’s high self-control “may interfere with being perceived as good company during leisure time and at parties” [44].
Overall, perceptions of self-control play an important role in how trustworthy a person is considered and the ramified positive and negative outcomes. While this connection is recognized across relationship types, distinct differences remain between their contexts. Naturally, organizational and work relationships are unique from social relationships. Social relationships are less formal and usually focus on personal connection, while organizational relationships often revolve around productivity and teamwork [50, 51]. These divergent characteristics likely contribute to how self-control and trust are understood in the respective relationships, breeding the variation of outcomes across contexts.
The unique findings from past research on self-control create an interesting paradigm for its relationship with trust and their related outcomes. Diving deeper into this relationship, we ask, what else might impact self-control’s signal of trustworthiness, and what would it mean?
Thus far, we have discussed the self-control and trust relationship in a quantitative sense, examining the extent to which high self-control is associated with greater perceptions of trust. While informative, a holistic understanding beyond the intensity of the relationship remains to be studied, and it requires us to better understand how and when the relationship occurs. We propose that the less explored, qualitative differences underlying self-control may impact the dynamic of the self-control and trust relationship, along with its potential outcomes. Here we offer our ideas and some relevant questions for future research.
As discussed above, self-control is conceptualized by choosing to pursue higher-order goals over lower-order goals [29]. Considering this definition, it is important to understand
One popular framework of goal content has been agentic and communal. Agentic goals are pursued in an effort to improve or satisfy oneself, while communal goals relate to the more interpersonal and connected pursuits of the person [56]. Agentic goals could include working out more to lose weight and look fit or reading more to grow your knowledge on different subjects. The person’s level of success in pursuing these goals can signal their competence or ability to others. Separately, communal goals could include being more proactive in reaching out to others to be a better friend or working hard to make extra money and better support your family. These goals can signal a person’s benevolence, or care and interest in others. These different types of goals serve varied functions in our lives, therefore, the content of a goal is important for the message it translates. The type of goal that is pursued can foster different perceptions of a person’s self-control abilities, even if the intensity of the person’s regulation is the same across the varied goals.
If someone is successful in pursuing their agentic goals it will signal high competence and ability in the person. This will subsequently act as a signal for cognition-based trust, which is partially formed from perceptions of ability, and thus cognition-based trust perceptions will increase towards the person. For example, someone that studies for an extra 5 hours during the week may be viewed as highly capable of improving their GPA, and thus more trustworthy in situations that test ability, which could lead to positive downstream outcomes like increased peer acceptance and better school adjustment [26].
In contrast, someone that is successful in pursuing their communal goals, which are based on interpersonal connection and care for others, will signal their high benevolence, promoting perceptions of affect-based trust. For example, a person that dedicates 5 hours a week to calling their family members to catch up may be seen as very caring, which would signal their affect-based trustworthiness. This would likely lead to positive outcomes for that person such as more friendships and greater acceptance from their peers [26, 27, 28].
While the promotion of cognitive-based or affect-based trust is likely beneficial to the perceived person, some recent research suggests that perceptions of high self-control can lead to negative outcomes as well, and those could be the result of a differential activation of the two kinds of trust. In one study, those viewed as high in self-control were seen as more “robot-like,” more competent, and less warm than those perceived as lower in self-control [42]. Those perceptions of high ability and competence would likely foster cognition-based but not affective-based trustworthiness in the perceived person. This asymmetry or lack of perceived benevolence (or warmth) then explains downstream negative social outcomes (e.g., reduced interest in socially connecting with the person) [45]. Additionally, this suggests a possible remedy that the presence of affect-based trust would act as a buffer to the negative outcomes. Fostering affect-based trust through successful self-control of communal goal pursuits, in addition to the already present cognition-based trust, may eliminate the negative outcomes that can be observed from perceptions of high self-control.
Overall, we propose that the goal content (e.g., agentic or communal) has an important impact on the formation of trust perceptions due to self-control, which can potentially lead to both positive and negative outcomes for the perceived. This proposition gives rise to new questions for the study of the relationship between self-control and trust perceptions. We have suggested that the negative social outcomes identified in recent research may be corrected by the addition of affect-based trust perceptions formed from successful communal goal pursuits. Alternatively, would relationships that demonstrate high self-control through only communal goals, producing solely affect-based trust perceptions, also result in negative outcomes? If so, would these consequences be exclusively agentic, and what would they entail? Additionally, how does the ratio of agentic and communal goal success relate to the formation of trust perceptions, and does this differ based on the context of the relationship (e.g., co-workers vs. romantic partners)? Finally, how would failed agentic or communal goal pursuits affect the outcomes of trust perceptions? These questions offer interesting potential avenues for future research.
An additional interesting qualitative factor to consider in the self-control and trust relationship is how differences in beliefs about self-control, in general, can impact perceptions of trustworthiness.
Lay theories of self-control recognize views that are commonly held about a person’s ability to self-regulate. Lay theories, or mindsets, are developed from our socialization and past experiences, and different types of lay theories have been identified in the self-control literature. The first relates to the belief that a person generally has a limited (slowly replenishing) or nonlimited (quickly replenishing) amount of self-control [57]. For example, if an individual successfully demonstrated their self-control abilities, someone with a limited self-control mindset would believe that the person no longer has their full capacity for implementing self-control, and that it will take time to be completely restored. Alternatively, a person with a nonlimited mindset would believe that an individual who demonstrated their self-control ability would have the same full capacity for self-control before their implementation of it, as well as quickly after.
This difference in mindset may create an important nuance for self-control’s relationship with trust perceptions. If a person holds a limited mindset about self-control abilities, they would believe that once an individual exhibits successful self-control, they will be less capable of successfully implementing self-control in subsequent tasks, as they have already used up some of their resource. While this would likely increase self-control perceptions for the already completed task, it may reduce expectations for the person’s future self-control abilities. In other words, the perceiver may have weaker trust perceptions because they expect the person to fail in demonstrating self-control in subsequent tasks, if there is not adequate time for their self-control abilities to replenish. An individual with a nonlimited mindset, however, would likely have greater trust perceptions, as they believe the person who just demonstrated successful self-control will have the same full capacity to do so in all subsequent self-control conflicts. This would likely lead to more positive outcomes for the perceived person.
Future research should test this idea, by investigating if those with limited self-control mindsets view others as less trustworthy after successfully demonstrating self-control. Other interesting questions remain as well, such as “How much time is needed for self-control abilities to replenish?” “Would the perceived person face negative outcomes from reduced trust perceptions?” “Do the types of goals pursued in the self-control action ‘use up’ one’s self-control reserves differently?” “Is self-control for agentic goals different from self-control for communal goals?” Lastly, “Would trustworthiness be reduced overall, or would perceptions of affect-based and cognition-based trust be impacted independently?”
The second type of mindset related to self-control focuses on the trait’s plasticity. It consists of a fixed (stable and unchanging) or malleable (varied and mutable) mindset [58]. Fixed vs. malleable mindset affects dispositional judgments [59]. Someone with a fixed mindset of self-control would believe that the amount of self-control displayed by a person in a particular situation represents their overall self-control abilities. Conversely, someone with a malleable mindset would believe that a person’s self-control abilities are susceptible to change, and therefore, a single instance that demonstrates self-control may not be indicative of the person’s abilities overall.
Similar to limited and nonlimited mindset, the assumptions that a person’s capacity for self-control will, or will not, change could color perceptions of the person’s trustworthiness. For instance, dispositionism in social judgments can be a double-edged sword, depending on the valence of first impressions. Those with a fixed mindset are more likely to believe that a person holds the same amount of self-control across different conflicts, and they would likely base their self-control perceptions off their first impressions of the perceived person’s self-control abilities. Thus, if they initially view a person to have low self-control, they may then see the person as untrustworthy overall. However, if the person is initially seen as high in self-control, they may then view them as an overall trustworthy person. Hence, the timing of the self-control incidence matters, and especially so for those with a fixed mindset of self-control.
The consideration of fixed and malleable mindset in relation to self-control trust perceptions breeds additional important questions. Primarily, since self-control abilities will likely fluctuate at some point, what does this mean for those with a fixed self-control mindset? What effect does a ‘slip up’ have on previously formed perceptions of self-control and trust? In relation to agentic and communal goals, would self-control perceptions formed by one of the goal types translate to assumptions for the other goal type? And regarding those with malleable mindsets, how strong can trust perceptions be if it is understood that one’s capacity for self-control is able to change?
Another way people may conceptualize self-control is in the materialization of their self-control efforts. In the process of pursuing a higher-order and distal goal over a lower-order and proximal goal, one may choose to utilize their willpower to effortfully inhibit the desire and temptation of the proximal goal. Another route the person could take would be to use strategies that allow them to proactively reduce their exposure to, and impact of, the temptation [29]. This can be done through manipulating the situation itself, such as selecting to be in an environment where the desire is not apparent (situation selection), or modifying the situation so it is easier to overcome the temptation (situation modification). Other strategies focus on altering the responses to temptations, such as directing focus away from the desire (attentional deployment) or manipulating the way we think about it, so it becomes less appealing (cognitive change) [60].
Research has shown that both types of strategies are used in self-control conflicts, however, there are mixed findings related to the success and prominence of these different methods [61, 62, 63]. Since both these dimensions of self-control implementation—willpower and strategies—show a range of conflicting results, it is understood that the way people demonstrate self-control can vary, and this could be due to their self-control beliefs. Some people may have a willpower-based mindset where they rely on effortful inhibition to overcome desire, while others could have a strategy-based mindset and utilize one or more strategies in their self-control efforts. The way a person thinks about self-control the method(s) of implementing it could influence how they perceive other’s self-control abilities.
The consideration of willpower-based and strategy-based mindsets in relation to self-control and trust perceptions sprouts several important questions. How does demonstrating control over oneself (i.e., willpower), compared to controlling the environment (i.e., using strategies), impact how trustworthy a person is viewed? Some research has suggested a timeline for when strategies and willpower are implemented in self-control conflicts. It is suggested that situational self-control strategies (i.e., situation selection and situation modification) are used first, followed by intrapsychic strategies (i.e., attentional deployment and cognitive change) [61]. Willpower, also referred to as response modulation, offers the final opportunity to overcome the desire. Since willpower can be considered the “last line of defense” in resisting a temptation, would a person that demonstrates self-control through effortful inhibition (i.e., willpower) be considered less trustworthy, as they could only overcome the desire in their final opportunity to do so? Or, would a person that demonstrates self-control through the use of strategies be considered less trustworthy, as the opportunity to change one’s environment may not always be present? Since one’s environment is more susceptible to change than the person themself, would someone that demonstrates strategy-based self-control be less reliable, and therefore less trustworthy than a person who demonstrates willpower-based self-control?
These questions are important for future research on self-control perceptions and their subsequent effects on trustworthiness. Willpower-based and strategy-based self-control mindsets may also lead to implications for the downstream outcomes of trustworthiness. Future research should examine the potential effects of willpower and strategy-based mindset, along with limited (nonlimited) and fixed (malleable) mindsets, on self-control and trust perceptions to increase insight into the relationship and its related outcomes.
Research has identified that self-control is an important predictor of trustworthiness. In considering the quantitative factors between self-control and trust, the relationship is almost exclusively positive, where greater self-control perceptions lead to increased perceptions of trustworthiness, which result in positive downstream outcomes. However, when considering the less researched potential qualitative factors that can impact the relationship, such as goal content and mindset, the connection between the traits and their subsequent outcomes becomes much more nuanced. This suggests that, future research should examine the impacts of goal content and mindset on the self-control and trust relationship, as well as their (positive and negative) downstream effects in order to form a more holistic understanding of self-control’s relationship with trust.
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Delac received his B.Sc.E.E. degree in 2003 and is currentlypursuing a Ph.D. degree at the University of Zagreb, Faculty of Electrical Engineering andComputing. His current research interests are digital image analysis, pattern recognition andbiometrics.",institutionString:null,institution:{name:"University of Zagreb",country:{name:"Croatia"}}},{id:"557",title:"Dr.",name:"Andon",middleName:"Venelinov",surname:"Topalov",slug:"andon-topalov",fullName:"Andon Topalov",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/557/images/1927_n.jpg",biography:"Dr. Andon V. Topalov received the MSc degree in Control Engineering from the Faculty of Information Systems, Technologies, and Automation at Moscow State University of Civil Engineering (MGGU) in 1979. He then received his PhD degree in Control Engineering from the Department of Automation and Remote Control at Moscow State Mining University (MGSU), Moscow, in 1984. 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These approaches are implemented on identifying human activities including normal movements of typical subjects and disorder-related movements such as stereotypical motor movements of autistic subjects. Experimental results show that our approaches improve performance of time series classification.",book:{id:"8362",slug:"time-series-analysis-data-methods-and-applications",title:"Time Series Analysis",fullTitle:"Time Series Analysis - Data, Methods, and Applications"},signatures:"Lamyaa Sadouk",authors:[{id:"257943",title:"Ph.D.",name:"Lamyaa",middleName:null,surname:"Sadouk",slug:"lamyaa-sadouk",fullName:"Lamyaa Sadouk"}]},{id:"56653",doi:"10.5772/intechopen.70230",title:"Bayesian Hypothesis Testing: An Alternative to Null Hypothesis Significance Testing (NHST) in Psychology and Social Sciences",slug:"bayesian-hypothesis-testing-an-alternative-to-null-hypothesis-significance-testing-nhst-in-psycholog",totalDownloads:3428,totalCrossrefCites:14,totalDimensionsCites:19,abstract:"Since the mid-1950s, there has been a clear predominance of the Frequentist approach to hypothesis testing, both in psychology and in social sciences. Despite its popularity in the field of statistics, Bayesian inference is barely known and used in psychology. Frequentist inference, and its null hypothesis significance testing (NHST), has been hegemonic through most of the history of scientific psychology. However, the NHST has not been exempt of criticisms. Therefore, the aim of this chapter is to introduce a Bayesian approach to hypothesis testing that may represent a useful complement, or even an alternative, to the current NHST. The advantages of this Bayesian approach over Frequentist NHST will be presented, providing examples that support its use in psychology and social sciences. Conclusions are outlined.",book:{id:"5964",slug:"bayesian-inference",title:"Bayesian Inference",fullTitle:"Bayesian Inference"},signatures:"Alonso Ortega and Gorka Navarrete",authors:[{id:"203438",title:"Dr.",name:"Alonso",middleName:null,surname:"Ortega",slug:"alonso-ortega",fullName:"Alonso Ortega"},{id:"208842",title:"Dr.",name:"Gorka",middleName:null,surname:"Navarrete",slug:"gorka-navarrete",fullName:"Gorka Navarrete"}]},{id:"59936",doi:"10.5772/intechopen.75007",title:"Application of Principal Component Analysis to Image Compression",slug:"application-of-principal-component-analysis-to-image-compression",totalDownloads:1819,totalCrossrefCites:12,totalDimensionsCites:15,abstract:"In this chapter, an introduction to the basics of principal component analysis (PCA) is given, aimed at presenting PCA applications to image compression. Here, concepts of linear algebra used in PCA are introduced, and PCA theoretical foundations are explained in connection with those concepts. Next, an image is compressed by using different principal components, and concepts such as image dimension reduction and image reconstruction quality are explained. Also, using the almost periodicity of the first principal component, a quality comparative analysis of a compressed image using two and eight principal components is carried out. 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Experimentation is an application of treatments applied to experimental units and is then part of a scientific method based on the measurement of one or more responses. It is necessary to observe the process and the operation of the system well. For this reason, in order to obtain a final result, an experimenter must plan and design experiments and analyzes the results. One of the most commonly used experimental designs for optimization is the response surface methodology (RSM). Because it allows evaluating the effects of multiple factors and their interactions on one or more response variables it is a useful method. In this section, recent studies have been compiled which aim to extraction of plant material in high yield and quality and determine optimum conditions for this extraction process.",book:{id:"5856",slug:"statistical-approaches-with-emphasis-on-design-of-experiments-applied-to-chemical-processes",title:"Statistical Approaches With Emphasis on Design of Experiments Applied to Chemical Processes",fullTitle:"Statistical Approaches With Emphasis on Design of Experiments Applied to Chemical Processes"},signatures:"Alev Yüksel Aydar",authors:[{id:"218870",title:"Dr.",name:"Alev Yüksel",middleName:null,surname:"Aydar",slug:"alev-yuksel-aydar",fullName:"Alev Yüksel Aydar"}]},{id:"60864",title:"Statistical Methodology for Evaluating Business Cycles with the Conditions of Their Synchronization and Harmonization",slug:"statistical-methodology-for-evaluating-business-cycles-with-the-conditions-of-their-synchronization-",totalDownloads:1357,totalCrossrefCites:1,totalDimensionsCites:2,abstract:"The importance of the topic of business cycle research and their interaction is due to the fact that the cyclical nature of development is a universal feature of the market economy (regardless of the level of development of the country’s economy and the principles of its organization). In all cases, cyclical ups and downs depend not only on internal system cyclical processes and their factors in countries but also on the consequences of intercountry interaction. The ability to measure and predict business cycles, taking into account their mutual influence, is a prerequisite for the development of an adequate business policy of countries and their associations.",book:{id:"6703",slug:"statistics-growing-data-sets-and-growing-demand-for-statistics",title:"Statistics",fullTitle:"Statistics - Growing Data Sets and Growing Demand for Statistics"},signatures:"Elena Zarova",authors:null},{id:"60246",title:"Statistical Research of Investment Appeal of Russian Regions",slug:"statistical-research-of-investment-appeal-of-russian-regions",totalDownloads:1012,totalCrossrefCites:0,totalDimensionsCites:0,abstract:"In this chapter, the methodological results directed on realization statistical research of investment appeal of Russian regions are offered. 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It is a statically approach where we develop the mathematical models through experimental trial runs to predict the possible output on the basis of the given input data or parameters. The aim of this chapter is to stimulate the engineering community to apply Taguchi technique to experimentation, the design of experiments, and to tackle quality problems in industrial chemical processes that they deal with. Based on years of research and applications, Dr. G. Taguchi has standardized the methods for each of these DOE application steps. Thus, DOE using Taguchi approach has become a much more attractive tool to practicing engineers and scientists. And since the last four decades, there were limitations when conventional experimental design techniques were applied to industrial experimentation. And Taguchi, also known as orthogonal array design, adds a new dimension to conventional experimental design. 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He pursued his postdoctoral studies at Rutgers University Medical School and the National Institutes of Health (NIH/NIDDK), USA. His research focuses on biochemistry, biophysics, genetics, molecular biology, and molecular medicine with specialization in the fields of drug design, protein structure-function, protein folding, prions, microRNA, pseudogenes, molecular cancer, epigenetics, metabolites, proteomics, genomics, protein expression, and characterization by spectroscopic and calorimetric methods.",institutionString:"University of Health Sciences",institution:null},{id:"180528",title:"Dr.",name:"Hiroyuki",middleName:null,surname:"Kagechika",slug:"hiroyuki-kagechika",fullName:"Hiroyuki Kagechika",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/180528/images/system/180528.jpg",biography:"Hiroyuki Kagechika received his bachelor’s degree and Ph.D. in Pharmaceutical Sciences from the University of Tokyo, Japan, where he served as an associate professor until 2004. He is currently a professor at the Institute of Biomaterials and Bioengineering (IBB), Tokyo Medical and Dental University (TMDU). From 2010 to 2012, he was the dean of the Graduate School of Biomedical Science. Since 2012, he has served as the vice dean of the Graduate School of Medical and Dental Sciences. He has been the director of the IBB since 2020. Dr. Kagechika’s major research interests are the medicinal chemistry of retinoids, vitamins D/K, and nuclear receptors. He has developed various compounds including a drug for acute promyelocytic leukemia.",institutionString:"Tokyo Medical and Dental University",institution:{name:"Tokyo Medical and Dental University",country:{name:"Japan"}}},{id:"94311",title:"Prof.",name:"Martins",middleName:"Ochubiojo",surname:"Ochubiojo Emeje",slug:"martins-ochubiojo-emeje",fullName:"Martins Ochubiojo Emeje",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/94311/images/system/94311.jpeg",biography:"Martins Emeje obtained a BPharm with distinction from Ahmadu Bello University, Nigeria, and an MPharm and Ph.D. from the University of Nigeria (UNN), where he received the best Ph.D. award and was enlisted as UNN’s “Face of Research.” He established the first nanomedicine center in Nigeria and was the pioneer head of the intellectual property and technology transfer as well as the technology innovation and support center. Prof. Emeje’s several international fellowships include the prestigious Raman fellowship. He has published more than 150 articles and patents. He is also the head of R&D at NIPRD and holds a visiting professor position at Nnamdi Azikiwe University, Nigeria. He has a postgraduate certificate in Project Management from Walden University, Minnesota, as well as a professional teaching certificate and a World Bank certification in Public Procurement. Prof. Emeje was a national chairman of academic pharmacists in Nigeria and the 2021 winner of the May & Baker Nigeria Plc–sponsored prize for professional service in research and innovation.",institutionString:"National Institute for Pharmaceutical Research and Development",institution:{name:"National Institute for Pharmaceutical Research and Development",country:{name:"Nigeria"}}},{id:"436430",title:"Associate Prof.",name:"Mesut",middleName:null,surname:"Işık",slug:"mesut-isik",fullName:"Mesut Işık",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/436430/images/19686_n.jpg",biography:null,institutionString:null,institution:{name:"Bilecik University",country:{name:"Turkey"}}},{id:"268659",title:"Ms.",name:"Xianquan",middleName:null,surname:"Zhan",slug:"xianquan-zhan",fullName:"Xianquan Zhan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/268659/images/8143_n.jpg",biography:"Dr. Zhan received his undergraduate and graduate training in the fields of preventive medicine and epidemiology and statistics at the West China University of Medical Sciences in China during 1989 to 1999. He received his post-doctoral training in oncology and cancer proteomics for two years at the Cancer Research Institute of Human Medical University in China. In 2001, he went to the University of Tennessee Health Science Center (UTHSC) in USA, where he was a post-doctoral researcher and focused on mass spectrometry and cancer proteomics. Then, he was appointed as an Assistant Professor of Neurology, UTHSC in 2005. He moved to the Cleveland Clinic in USA as a Project Scientist/Staff in 2006 where he focused on the studies of eye disease proteomics and biomarkers. He returned to UTHSC as an Assistant Professor of Neurology in the end of 2007, engaging in proteomics and biomarker studies of lung diseases and brain tumors, and initiating the studies of predictive, preventive, and personalized medicine (PPPM) in cancer. In 2010, he was promoted to Associate Professor of Neurology, UTHSC. Currently, he is a Professor at Xiangya Hospital of Central South University in China, Fellow of Royal Society of Medicine (FRSM), the European EPMA National Representative in China, Regular Member of American Association for the Advancement of Science (AAAS), European Cooperation of Science and Technology (e-COST) grant evaluator, Associate Editors of BMC Genomics, BMC Medical Genomics, EPMA Journal, and Frontiers in Endocrinology, Executive Editor-in-Chief of Med One. He has\npublished 116 peer-reviewed research articles, 16 book chapters, 2 books, and 2 US patents. His current main research interest focuses on the studies of cancer proteomics and biomarkers, and the use of modern omics techniques and systems biology for PPPM in cancer, and on the development and use of 2DE-LC/MS for the large-scale study of human proteoforms.",institutionString:null,institution:{name:"Xiangya Hospital Central South University",country:{name:"China"}}},{id:"40482",title:null,name:"Rizwan",middleName:null,surname:"Ahmad",slug:"rizwan-ahmad",fullName:"Rizwan Ahmad",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/40482/images/system/40482.jpeg",biography:"Dr. Rizwan Ahmad is a University Professor and Coordinator, Quality and Development, College of Medicine, Imam Abdulrahman bin Faisal University, Saudi Arabia. Previously, he was Associate Professor of Human Function, Oman Medical College, Oman, and SBS University, Dehradun. Dr. Ahmad completed his education at Aligarh Muslim University, Aligarh. He has published several articles in peer-reviewed journals, chapters, and edited books. His area of specialization is free radical biochemistry and autoimmune diseases.",institutionString:"Imam Abdulrahman Bin Faisal University",institution:{name:"Imam Abdulrahman Bin Faisal University",country:{name:"Saudi Arabia"}}},{id:"41865",title:"Prof.",name:"Farid A.",middleName:null,surname:"Badria",slug:"farid-a.-badria",fullName:"Farid A. Badria",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/41865/images/system/41865.jpg",biography:"Farid A. Badria, Ph.D., is the recipient of several awards, including The World Academy of Sciences (TWAS) Prize for Public Understanding of Science; the World Intellectual Property Organization (WIPO) Gold Medal for best invention; Outstanding Arab Scholar, Kuwait; and the Khwarizmi International Award, Iran. He has 250 publications, 12 books, 20 patents, and several marketed pharmaceutical products to his credit. He continues to lead research projects on developing new therapies for liver, skin disorders, and cancer. Dr. Badria was listed among the world’s top 2% of scientists in medicinal and biomolecular chemistry in 2019 and 2020. He is a member of the Arab Development Fund, Kuwait; International Cell Research Organization–United Nations Educational, Scientific and Cultural Organization (ICRO–UNESCO), Chile; and UNESCO Biotechnology France",institutionString:"Mansoura University",institution:{name:"Mansoura University",country:{name:"Egypt"}}},{id:"329385",title:"Dr.",name:"Rajesh K.",middleName:"Kumar",surname:"Singh",slug:"rajesh-k.-singh",fullName:"Rajesh K. Singh",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329385/images/system/329385.png",biography:"Dr. Singh received a BPharm (2003) and MPharm (2005) from Panjab University, Chandigarh, India, and a Ph.D. (2013) from Punjab Technical University (PTU), Jalandhar, India. He has more than sixteen years of teaching experience and has supervised numerous postgraduate and Ph.D. students. He has to his credit more than seventy papers in SCI- and SCOPUS-indexed journals, fifty-five conference proceedings, four books, six Best Paper Awards, and five projects from different government agencies. He is currently an editorial board member of eight international journals and a reviewer for more than fifty scientific journals. He received Top Reviewer and Excellent Peer Reviewer Awards from Publons in 2016 and 2017, respectively. He is also on the panel of The International Reviewer for reviewing research proposals for grants from the Royal Society. He also serves as a Publons Academy mentor and Bentham brand ambassador.",institutionString:"Punjab Technical University",institution:{name:"Punjab Technical University",country:{name:"India"}}},{id:"142388",title:"Dr.",name:"Thiago",middleName:"Gomes",surname:"Gomes Heck",slug:"thiago-gomes-heck",fullName:"Thiago Gomes Heck",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/142388/images/7259_n.jpg",biography:null,institutionString:null,institution:{name:"Universidade Regional do Noroeste do Estado do Rio Grande do Sul",country:{name:"Brazil"}}},{id:"336273",title:"Assistant Prof.",name:"Janja",middleName:null,surname:"Zupan",slug:"janja-zupan",fullName:"Janja Zupan",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/336273/images/14853_n.jpeg",biography:"Janja Zupan graduated in 2005 at the Department of Clinical Biochemistry (superviser prof. dr. Janja Marc) in the field of genetics of osteoporosis. Since November 2009 she is working as a Teaching Assistant at the Faculty of Pharmacy, Department of Clinical Biochemistry. In 2011 she completed part of her research and PhD work at Institute of Genetics and Molecular Medicine, University of Edinburgh. She finished her PhD entitled The influence of the proinflammatory cytokines on the RANK/RANKL/OPG in bone tissue of osteoporotic and osteoarthritic patients in 2012. From 2014-2016 she worked at the Institute of Biomedical Sciences, University of Aberdeen as a postdoctoral research fellow on UK Arthritis research project where she gained knowledge in mesenchymal stem cells and regenerative medicine. She returned back to University of Ljubljana, Faculty of Pharmacy in 2016. She is currently leading project entitled Mesenchymal stem cells-the keepers of tissue endogenous regenerative capacity facing up to aging of the musculoskeletal system funded by Slovenian Research Agency.",institutionString:null,institution:{name:"University of Ljubljana",country:{name:"Slovenia"}}},{id:"357453",title:"Dr.",name:"Radheshyam",middleName:null,surname:"Maurya",slug:"radheshyam-maurya",fullName:"Radheshyam Maurya",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/357453/images/16535_n.jpg",biography:null,institutionString:null,institution:{name:"University of Hyderabad",country:{name:"India"}}},{id:"418340",title:"Dr.",name:"Jyotirmoi",middleName:null,surname:"Aich",slug:"jyotirmoi-aich",fullName:"Jyotirmoi Aich",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000038Ugi5QAC/Profile_Picture_2022-04-15T07:48:28.png",biography:"Biotechnologist with 15 years of research including 6 years of teaching experience. Demonstrated record of scientific achievements through consistent publication record (H index = 13, with 874 citations) in high impact journals such as Nature Communications, Oncotarget, Annals of Oncology, PNAS, and AJRCCM, etc. Strong research professional with a post-doctorate from ACTREC where I gained experimental oncology experience in clinical settings and a doctorate from IGIB where I gained expertise in asthma pathophysiology. A well-trained biotechnologist with diverse experience on the bench across different research themes ranging from asthma to cancer and other infectious diseases. An individual with a strong commitment and innovative mindset. Have the ability to work on diverse projects such as regenerative and molecular medicine with an overall mindset of improving healthcare.",institutionString:"DY Patil Deemed to Be University",institution:null},{id:"349288",title:"Prof.",name:"Soumya",middleName:null,surname:"Basu",slug:"soumya-basu",fullName:"Soumya Basu",position:null,profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y000035QxIDQA0/Profile_Picture_2022-04-15T07:47:01.jpg",biography:"Soumya Basu, Ph.D., is currently working as an Associate Professor at Dr. D. Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pune, Maharashtra, India. With 16+ years of trans-disciplinary research experience in Drug Design, development, and pre-clinical validation; 20+ research article publications in journals of repute, 9+ years of teaching experience, trained with cross-disciplinary education, Dr. Basu is a life-long learner and always thrives for new challenges.\r\nHer research area is the design and synthesis of small molecule partial agonists of PPAR-γ in lung cancer. She is also using artificial intelligence and deep learning methods to understand the exosomal miRNA’s role in cancer metastasis. Dr. Basu is the recipient of many awards including the Early Career Research Award from the Department of Science and Technology, Govt. of India. She is a reviewer of many journals like Molecular Biology Reports, Frontiers in Oncology, RSC Advances, PLOS ONE, Journal of Biomolecular Structure & Dynamics, Journal of Molecular Graphics and Modelling, etc. She has edited and authored/co-authored 21 journal papers, 3 book chapters, and 15 abstracts. She is a Board of Studies member at her university. She is a life member of 'The Cytometry Society”-in India and 'All India Cell Biology Society”- in India.",institutionString:"Dr. D.Y. Patil Vidyapeeth, Pune",institution:{name:"Dr. D.Y. Patil Vidyapeeth, Pune",country:{name:"India"}}},{id:"354817",title:"Dr.",name:"Anubhab",middleName:null,surname:"Mukherjee",slug:"anubhab-mukherjee",fullName:"Anubhab Mukherjee",position:null,profilePictureURL:"https://intech-files.s3.amazonaws.com/0033Y0000365PbRQAU/ProfilePicture%202022-04-15%2005%3A11%3A18.480",biography:"A former member of Laboratory of Nanomedicine, Brigham and Women’s Hospital, Harvard University, Boston, USA, Dr. Anubhab Mukherjee is an ardent votary of science who strives to make an impact in the lives of those afflicted with cancer and other chronic/acute ailments. He completed his Ph.D. from CSIR-Indian Institute of Chemical Technology, Hyderabad, India, having been skilled with RNAi, liposomal drug delivery, preclinical cell and animal studies. He pursued post-doctoral research at College of Pharmacy, Health Science Center, Texas A & M University and was involved in another postdoctoral research at Department of Translational Neurosciences and Neurotherapeutics, John Wayne Cancer Institute, Santa Monica, California. In 2015, he worked in Harvard-MIT Health Sciences & Technology as a visiting scientist. He has substantial experience in nanotechnology-based formulation development and successfully served various Indian organizations to develop pharmaceuticals and nutraceutical products. He is an inventor in many US patents and an author in many peer-reviewed articles, book chapters and books published in various media of international repute. Dr. Mukherjee is currently serving as Principal Scientist, R&D at Esperer Onco Nutrition (EON) Pvt. Ltd. and heads the Hyderabad R&D center of the organization.",institutionString:"Esperer Onco Nutrition Pvt Ltd.",institution:null},{id:"319365",title:"Assistant Prof.",name:"Manash K.",middleName:null,surname:"Paul",slug:"manash-k.-paul",fullName:"Manash K. Paul",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/319365/images/system/319365.png",biography:"Manash K. Paul is a Principal Investigator and Scientist at the University of California Los Angeles. He has contributed significantly to the fields of stem cell biology, regenerative medicine, and lung cancer. His research focuses on various signaling processes involved in maintaining stem cell homeostasis during the injury-repair process, deciphering lung stem cell niche, pulmonary disease modeling, immuno-oncology, and drug discovery. He is currently investigating the role of extracellular vesicles in premalignant lung cell migration and detecting the metastatic phenotype of lung cancer via machine-learning-based analyses of exosomal signatures. Dr. Paul has published in more than fifty peer-reviewed international journals and is highly cited. He is the recipient of many awards, including the UCLA Vice Chancellor’s award, a senior member of the Institute of Electrical and Electronics Engineers (IEEE), and an editorial board member for several international journals.",institutionString:"University of California Los Angeles",institution:{name:"University of California Los Angeles",country:{name:"United States of America"}}},{id:"311457",title:"Dr.",name:"Júlia",middleName:null,surname:"Scherer Santos",slug:"julia-scherer-santos",fullName:"Júlia Scherer Santos",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/311457/images/system/311457.jpg",biography:"Dr. Júlia Scherer Santos works in the areas of cosmetology, nanotechnology, pharmaceutical technology, beauty, and aesthetics. Dr. Santos also has experience as a professor of graduate courses. Graduated in Pharmacy, specialization in Cosmetology and Cosmeceuticals applied to aesthetics, specialization in Aesthetic and Cosmetic Health, and a doctorate in Pharmaceutical Nanotechnology. Teaching experience in Pharmacy and Aesthetics and Cosmetics courses. She works mainly on the following subjects: nanotechnology, cosmetology, pharmaceutical technology, aesthetics.",institutionString:"Universidade Federal de Juiz de Fora",institution:{name:"Universidade Federal de Juiz de Fora",country:{name:"Brazil"}}},{id:"219081",title:"Dr.",name:"Abdulsamed",middleName:null,surname:"Kükürt",slug:"abdulsamed-kukurt",fullName:"Abdulsamed Kükürt",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/219081/images/system/219081.png",biography:"Dr. Kükürt graduated from Uludağ University in Turkey. He started his academic career as a Research Assistant in the Department of Biochemistry at Kafkas University. In 2019, he completed his Ph.D. program in the Department of Biochemistry at the Institute of Health Sciences. He is currently working at the Department of Biochemistry, Kafkas University. He has 27 published research articles in academic journals, 11 book chapters, and 37 papers. He took part in 10 academic projects. He served as a reviewer for many articles. He still serves as a member of the review board in many academic journals. He is currently working on the protective activity of phenolic compounds in disorders associated with oxidative stress and inflammation.",institutionString:null,institution:{name:"Kafkas University",country:{name:"Turkey"}}},{id:"178366",title:"Dr.",name:"Volkan",middleName:null,surname:"Gelen",slug:"volkan-gelen",fullName:"Volkan Gelen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/178366/images/system/178366.jpg",biography:"Volkan Gelen is a Physiology specialist who received his veterinary degree from Kafkas University in 2011. Between 2011-2015, he worked as an assistant at Atatürk University, Faculty of Veterinary Medicine, Department of Physiology. In 2016, he joined Kafkas University, Faculty of Veterinary Medicine, Department of Physiology as an assistant professor. Dr. Gelen has been engaged in various academic activities at Kafkas University since 2016. There he completed 5 projects and has 3 ongoing projects. He has 60 articles published in scientific journals and 20 poster presentations in scientific congresses. His research interests include physiology, endocrine system, cancer, diabetes, cardiovascular system diseases, and isolated organ bath system studies.",institutionString:"Kafkas University",institution:{name:"Kafkas University",country:{name:"Turkey"}}},{id:"418963",title:"Dr.",name:"Augustine Ododo",middleName:"Augustine",surname:"Osagie",slug:"augustine-ododo-osagie",fullName:"Augustine Ododo Osagie",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/418963/images/16900_n.jpg",biography:"Born into the family of Osagie, a prince of the Benin Kingdom. I am currently an academic in the Department of Medical Biochemistry, University of Benin. Part of the duties are to teach undergraduate students and conduct academic research.",institutionString:null,institution:{name:"University of Benin",country:{name:"Nigeria"}}},{id:"192992",title:"Prof.",name:"Shagufta",middleName:null,surname:"Perveen",slug:"shagufta-perveen",fullName:"Shagufta Perveen",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/192992/images/system/192992.png",biography:"Prof. Shagufta Perveen is a Distinguish Professor in the Department of Pharmacognosy, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia. Dr. Perveen has acted as the principal investigator of major research projects funded by the research unit of King Saud University. She has more than ninety original research papers in peer-reviewed journals of international repute to her credit. She is a fellow member of the Royal Society of Chemistry UK and the American Chemical Society of the United States.",institutionString:"King Saud University",institution:{name:"King Saud University",country:{name:"Saudi Arabia"}}},{id:"49848",title:"Dr.",name:"Wen-Long",middleName:null,surname:"Hu",slug:"wen-long-hu",fullName:"Wen-Long Hu",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/49848/images/system/49848.jpg",biography:"Wen-Long Hu is Chief of the Division of Acupuncture, Department of Chinese Medicine at Kaohsiung Chang Gung Memorial Hospital, as well as an adjunct associate professor at Fooyin University and Kaohsiung Medical University. Wen-Long is President of Taiwan Traditional Chinese Medicine Medical Association. He has 28 years of experience in clinical practice in laser acupuncture therapy and 34 years in acupuncture. He is an invited speaker for lectures and workshops in laser acupuncture at many symposiums held by medical associations. He owns the patent for herbal preparation and producing, and for the supercritical fluid-treated needle. Dr. Hu has published three books, 12 book chapters, and more than 30 papers in reputed journals, besides serving as an editorial board member of repute.",institutionString:"Kaohsiung Chang Gung Memorial Hospital",institution:{name:"Kaohsiung Chang Gung Memorial Hospital",country:{name:"Taiwan"}}},{id:"298472",title:"Prof.",name:"Andrey V.",middleName:null,surname:"Grechko",slug:"andrey-v.-grechko",fullName:"Andrey V. Grechko",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/298472/images/system/298472.png",biography:"Andrey Vyacheslavovich Grechko, Ph.D., Professor, is a Corresponding Member of the Russian Academy of Sciences. He graduated from the Semashko Moscow Medical Institute (Semashko National Research Institute of Public Health) with a degree in Medicine (1998), the Clinical Department of Dermatovenerology (2000), and received a second higher education in Psychology (2009). Professor A.V. Grechko held the position of Сhief Physician of the Central Clinical Hospital in Moscow. He worked as a professor at the faculty and was engaged in scientific research at the Medical University. Starting in 2013, he has been the initiator of the creation of the Federal Scientific and Clinical Center for Intensive Care and Rehabilitology, Moscow, Russian Federation, where he also serves as Director since 2015. He has many years of experience in research and teaching in various fields of medicine, is an author/co-author of more than 200 scientific publications, 13 patents, 15 medical books/chapters, including Chapter in Book «Metabolomics», IntechOpen, 2020 «Metabolomic Discovery of Microbiota Dysfunction as the Cause of Pathology».",institutionString:"Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology",institution:null},{id:"199461",title:"Prof.",name:"Natalia V.",middleName:null,surname:"Beloborodova",slug:"natalia-v.-beloborodova",fullName:"Natalia V. Beloborodova",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/199461/images/system/199461.jpg",biography:'Natalia Vladimirovna Beloborodova was educated at the Pirogov Russian National Research Medical University, with a degree in pediatrics in 1980, a Ph.D. in 1987, and a specialization in Clinical Microbiology from First Moscow State Medical University in 2004. She has been a Professor since 1996. Currently, she is the Head of the Laboratory of Metabolism, a division of the Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology, Moscow, Russian Federation. N.V. Beloborodova has many years of clinical experience in the field of intensive care and surgery. She studies infectious complications and sepsis. She initiated a series of interdisciplinary clinical and experimental studies based on the concept of integrating human metabolism and its microbiota. Her scientific achievements are widely known: she is the recipient of the Marie E. Coates Award \\"Best lecturer-scientist\\" Gustafsson Fund, Karolinska Institutes, Stockholm, Sweden, and the International Sepsis Forum Award, Pasteur Institute, Paris, France (2014), etc. Professor N.V. Beloborodova wrote 210 papers, five books, 10 chapters and has edited four books.',institutionString:"Federal Research and Clinical Center of Intensive Care Medicine and Rehabilitology",institution:null},{id:"354260",title:"Ph.D.",name:"Tércio Elyan",middleName:"Azevedo",surname:"Azevedo Martins",slug:"tercio-elyan-azevedo-martins",fullName:"Tércio Elyan Azevedo Martins",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/354260/images/16241_n.jpg",biography:"Graduated in Pharmacy from the Federal University of Ceará with the modality in Industrial Pharmacy, Specialist in Production and Control of Medicines from the University of São Paulo (USP), Master in Pharmaceuticals and Medicines from the University of São Paulo (USP) and Doctor of Science in the program of Pharmaceuticals and Medicines by the University of São Paulo. Professor at Universidade Paulista (UNIP) in the areas of chemistry, cosmetology and trichology. Assistant Coordinator of the Higher Course in Aesthetic and Cosmetic Technology at Universidade Paulista Campus Chácara Santo Antônio. Experience in the Pharmacy area, with emphasis on Pharmacotechnics, Pharmaceutical Technology, Research and Development of Cosmetics, acting mainly on topics such as cosmetology, antioxidant activity, aesthetics, photoprotection, cyclodextrin and thermal analysis.",institutionString:null,institution:{name:"University of Sao Paulo",country:{name:"Brazil"}}},{id:"334285",title:"Ph.D. Student",name:"Sameer",middleName:"Kumar",surname:"Jagirdar",slug:"sameer-jagirdar",fullName:"Sameer Jagirdar",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/334285/images/14691_n.jpg",biography:"I\\'m a graduate student at the center for biosystems science and engineering at the Indian Institute of Science, Bangalore, India. I am interested in studying host-pathogen interactions at the biomaterial interface.",institutionString:null,institution:{name:"Indian Institute of Science Bangalore",country:{name:"India"}}},{id:"329248",title:"Dr.",name:"Md. Faheem",middleName:null,surname:"Haider",slug:"md.-faheem-haider",fullName:"Md. Faheem Haider",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329248/images/system/329248.jpg",biography:"Dr. Md. Faheem Haider completed his BPharm in 2012 at Integral University, Lucknow, India. In 2014, he completed his MPharm with specialization in Pharmaceutics at Babasaheb Bhimrao Ambedkar University, Lucknow, India. He received his Ph.D. degree from Jamia Hamdard University, New Delhi, India, in 2018. He was selected for the GPAT six times and his best All India Rank was 34. Currently, he is an assistant professor at Integral University. Previously he was an assistant professor at IIMT University, Meerut, India. He has experience teaching DPharm, Pharm.D, BPharm, and MPharm students. He has more than five publications in reputed journals to his credit. Dr. Faheem’s research area is the development and characterization of nanoformulation for the delivery of drugs to various organs.",institutionString:"Integral University",institution:{name:"Integral University",country:{name:"India"}}},{id:"329795",title:"Dr.",name:"Mohd Aftab",middleName:"Aftab",surname:"Siddiqui",slug:"mohd-aftab-siddiqui",fullName:"Mohd Aftab Siddiqui",position:null,profilePictureURL:"https://mts.intechopen.com/storage/users/329795/images/system/329795.png",biography:"Dr. Mohd Aftab Siddiqui is an assistant professor in the Faculty of Pharmacy, Integral University, Lucknow, India, where he obtained a Ph.D. in Pharmacology in 2020. He also obtained a BPharm and MPharm from the same university in 2013 and 2015, respectively. His area of research is the pharmacological screening of herbal drugs/natural products in liver cancer and cardiac diseases. He is a member of many professional bodies and has guided many MPharm and PharmD research projects. Dr. Siddiqui has many national and international publications and one German patent to his credit.",institutionString:"Integral University",institution:null}]}},subseries:{item:{id:"28",type:"subseries",title:"Animal Reproductive Biology and Technology",keywords:"Animal Reproduction, Artificial Insemination, Embryos, Cryopreservation, Conservation, Breeding, Epigenetics",scope:"The advances of knowledge on animal reproductive biology and technologies revolutionized livestock production. Artificial insemination, for example, was the first technology applied on a large scale, initially in dairy cattle and afterward applied to other species. Nowadays, embryo production and transfer are used commercially along with other technologies to modulate epigenetic regulation. 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Possible contributions can address (but are not limited to) the following research topics: Bioinspired design and control of exoskeletons, orthoses, and prostheses; Experimental evaluation of the effect of assistive devices (e.g., influence on gait, balance, and neuromuscular system); Bioinspired technologies for rehabilitation, including clinical studies reporting evaluations; Application of neuromuscular and biomechanical models to the development of bioinspired technology.',annualVolume:11404,isOpenForSubmission:!0,coverUrl:"https://cdn.intechopen.com/series_topics/covers/8.jpg",editor:{id:"144937",title:"Prof.",name:"Adriano",middleName:"De Oliveira",surname:"Andrade",fullName:"Adriano Andrade",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002bRC8QQAW/Profile_Picture_1625219101815",institutionString:null,institution:{name:"Federal University of Uberlândia",institutionURL:null,country:{name:"Brazil"}}},editorTwo:null,editorThree:null,editorialBoard:[{id:"49517",title:"Prof.",name:"Hitoshi",middleName:null,surname:"Tsunashima",fullName:"Hitoshi Tsunashima",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002aYTP4QAO/Profile_Picture_1625819726528",institutionString:null,institution:{name:"Nihon University",institutionURL:null,country:{name:"Japan"}}},{id:"425354",title:"Dr.",name:"Marcus",middleName:"Fraga",surname:"Vieira",fullName:"Marcus Vieira",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0033Y00003BJSgIQAX/Profile_Picture_1627904687309",institutionString:null,institution:{name:"Universidade Federal de Goiás",institutionURL:null,country:{name:"Brazil"}}},{id:"196746",title:"Dr.",name:"Ramana",middleName:null,surname:"Vinjamuri",fullName:"Ramana Vinjamuri",profilePictureURL:"https://mts.intechopen.com/storage/users/196746/images/system/196746.jpeg",institutionString:"University of Maryland, Baltimore County",institution:{name:"University of Maryland, Baltimore County",institutionURL:null,country:{name:"United States of America"}}}]},{id:"9",title:"Biotechnology - Biosensors, Biomaterials and Tissue Engineering",keywords:"Biotechnology, Biosensors, Biomaterials, Tissue Engineering",scope:"The Biotechnology - Biosensors, Biomaterials and Tissue Engineering topic within the Biomedical Engineering Series aims to rapidly publish contributions on all aspects of biotechnology, biosensors, biomaterial and tissue engineering. We encourage the submission of manuscripts that provide novel and mechanistic insights that report significant advances in the fields. Topics can include but are not limited to: Biotechnology such as biotechnological products and process engineering; Biotechnologically relevant enzymes and proteins; Bioenergy and biofuels; Applied genetics and molecular biotechnology; Genomics, transcriptomics, proteomics; Applied microbial and cell physiology; Environmental biotechnology; Methods and protocols. Moreover, topics in biosensor technology, like sensors that incorporate enzymes, antibodies, nucleic acids, whole cells, tissues and organelles, and other biological or biologically inspired components will be considered, and topics exploring transducers, including those based on electrochemical and optical piezoelectric, thermal, magnetic, and micromechanical elements. Chapters exploring biomaterial approaches such as polymer synthesis and characterization, drug and gene vector design, biocompatibility, immunology and toxicology, and self-assembly at the nanoscale, are welcome. Finally, the tissue engineering subcategory will support topics such as the fundamentals of stem cells and progenitor cells and their proliferation, differentiation, bioreactors for three-dimensional culture and studies of phenotypic changes, stem and progenitor cells, both short and long term, ex vivo and in vivo implantation both in preclinical models and also in clinical trials.",annualVolume:11405,isOpenForSubmission:!0,coverUrl:"https://cdn.intechopen.com/series_topics/covers/9.jpg",editor:{id:"126286",title:"Dr.",name:"Luis",middleName:"Jesús",surname:"Villarreal-Gómez",fullName:"Luis Villarreal-Gómez",profilePictureURL:"https://mts.intechopen.com/storage/users/126286/images/system/126286.jpg",institutionString:null,institution:{name:"Autonomous University of Baja California",institutionURL:null,country:{name:"Mexico"}}},editorTwo:null,editorThree:null,editorialBoard:[{id:"35539",title:"Dr.",name:"Cecilia",middleName:null,surname:"Cristea",fullName:"Cecilia Cristea",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002aYQ65QAG/Profile_Picture_1621007741527",institutionString:null,institution:{name:"Iuliu Hațieganu University of Medicine and Pharmacy",institutionURL:null,country:{name:"Romania"}}},{id:"40735",title:"Dr.",name:"Gil",middleName:"Alberto Batista",surname:"Gonçalves",fullName:"Gil Gonçalves",profilePictureURL:"https://s3.us-east-1.amazonaws.com/intech-files/0030O00002aYRLGQA4/Profile_Picture_1628492612759",institutionString:null,institution:{name:"University of Aveiro",institutionURL:null,country:{name:"Portugal"}}},{id:"211725",title:"Associate Prof.",name:"Johann F.",middleName:null,surname:"Osma",fullName:"Johann F. 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