\r\n\t2) Human sexual disorders in males and females.
\r\n\t3) Psychological aspects of the human sexual response cycle and its disorders.
\r\n\t4) The therapeutic aspects.
\r\n\tThe human sexual response cycle and human sexual behavior are interrelated. How this inter-relationship and its association to normal sexual health need to be delineated. In a world torn between sex and sexually transmitted disease, clear-cut scientific information in the form of a monograph is required to educate.
\r\n\r\n\tHuman sexuality, gender identity, and sexuo-erotic orientation play great roles in human health and disease. Sex education is the need of the hour and a reflection will be timely.
",isbn:"978-1-80355-151-7",printIsbn:"978-1-80355-150-0",pdfIsbn:"978-1-80355-152-4",doi:null,price:0,priceEur:0,priceUsd:0,slug:null,numberOfPages:0,isOpenForSubmission:!1,isSalesforceBook:!1,hash:"13af09c4cf93ae89789a3db597972cf6",bookSignature:"Dr. Dhastagir Sultan Sheriff",publishedDate:null,coverURL:"https://cdn.intechopen.com/books/images_new/11267.jpg",keywords:"Master and Johnson's Cycle, Sex Education, Premature Ejaculation, Orgasmic Disorders, Sexual Aversion Disorders, Dyspareunia, Vaginismus, Sex Hormones, Sexually Transmitted Diseases, Impotence, Low Libido, Blood Analyses",numberOfDownloads:99,numberOfWosCitations:0,numberOfCrossrefCitations:0,numberOfDimensionsCitations:0,numberOfTotalCitations:0,isAvailableForWebshopOrdering:!0,dateEndFirstStepPublish:"August 18th 2021",dateEndSecondStepPublish:"March 3rd 2022",dateEndThirdStepPublish:"May 2nd 2022",dateEndFourthStepPublish:"July 21st 2022",dateEndFifthStepPublish:"September 19th 2022",remainingDaysToSecondStep:"3 months",secondStepPassed:!0,currentStepOfPublishingProcess:4,editedByType:null,kuFlag:!1,biosketch:"Dr. Sheriff is a life counselor, sex educationist, and researcher with over 35 years of teaching experience, five authored books, and editorials written in the British Journal of Sexology and the Journal of Royal Society of Medicine. 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Each lobule is composed of 10–100 grapelike clusters of milk‐secreting glands termed acini, which are connected to lactiferous ducts [3]. The epithelium throughout the acini and ducts consists of two layers: an inner layer of polarized and cuboidal luminal cells that encapsulate a central lumen, and a basal outer layer of myoepithelial cells with contractile properties conferring these cells an active role in the milk excretion during lactation [3, 4]. Myoepithelial cells also ensure the maintenance of the adjacent luminal epithelial cell polarity and the synthesis of a laminin‐rich basement membrane (BM) that forms a structural barrier separating the glandular epithelium from the stroma [5]. In the normal mammary gland, luminal epithelial cells are characterized by the expression of the luminal cytokeratins CK7, CK8 and CK18, sialomucin, epithelial‐specific antigen, occludin and integrin β4 [5, 6]. On the other hand, myoepithelial cells express the basal cytokeratins CK5, CK14 and CK17 along with CD10/CALLA, alpha‐smooth actin and P63 [6, 7]. The stroma surrounding the mammary gland consists of an insoluble extracellular matrix (laminin, fibronectin, collagen, proteoglycans), mesenchymal cells (fibroblasts, adipocytes, endothelial cells and resident immune cells), and various growth factors and cytokines [8]. Aberrant interactions between mammary epithelial cells and the stroma may lead to structural and functional alterations of the mammary gland biology and ultimately promote breast malignancy [8].
Many suspicious mammograms or palpable findings turn out to be benign lesions following breast biopsy [9]. However, based on the histopathological report and family history, about 3–10% of these benign lesions are considered to be at high risk of later breast cancer and are referred to as atypical hyperplasia [10, 11]. Atypical hyperplasia is a premalignant lesion diagnosed based on the architectural pattern, cytology and the disease extent and is traditionally classified into two subtypes: atypical ductal hyperplasia (ADH) and atypical lobular hyperplasia (ALH) [12]. The absolute risk of developing breast cancer has been estimated at about 30% for women diagnosed with atypical hyperplasia after 25 years of follow‐up [13].
\n“In Situ” carcinoma, also known as stage 0 breast cancer, is defined by the clonal proliferation of neoplastic epithelial cells within the ducts (e.g., ductal carcinoma in situ) or the lobules (e.g., lobular carcinoma In Situ) of the mammary gland. “In Situ
Even if DCIS is not immediately life‐threatening, 14–53% of untreated DCIS lesions will progress to invasive ductal carcinoma (IDC) with considerable inconsistency in the timing and nature of this transition [16–20]. The most widely accepted model of breast carcinogenesis is the model of “linear” progression that hypothesizes that DCIS is an obligate precursor of IDC evolving through sequential stages, dependent upon early genetic and/or epigenetic changes [21–26].
Generally, surgery alone will reduce the risk of mortality following DCIS to less than 5% for lumpectomy and 1% for mastectomy [27]. Surgery followed by radiation and/or hormonal therapy may not alter overall survival dramatically but tends to reduce recurrence, and in the case of hormonal therapy, contralateral breast cancers [28–30]. Decisions regarding therapy are made on a case‐by‐case basis in function of the clinical presentation and patient choice. Though controversy remains surrounding treatment strategies, generally an argument with concerns of overtreatment voiced against fears of under‐treatment; it seems unlikely that current paradigms will change without further research and understanding into the natural history of DCIS progression and identification of clinically actionable markers of risk.
Using molecular profiling, Perou et al. first described the subtypes of human breast cancer generating four intrinsic subtypes: luminal A, luminal B (that are frequently ER (estrogen receptor) and PR (progesterone receptor) positive); HER2+ (human epidermal growth factor receptor); and basal‐like (frequently HER2‐,ER‐, PR‐ also known as triple negative) [31]. These subtypes are utilized currently to subcategorize breast cancer and have demonstrated clinical applicability as individual subtypes display differences in biology and behavior that inform prognosis and course of treatment [31–33].
\nIn 2008, Tamimi et al. performed tissue microarrays for demonstrably reliable [34–37] surrogates for the intrinsic subtypes identified by Perou et al. in 2000 in order to evaluate the prevalence of these phenotypes among DCIS cases. A large cohort of both DCIS and IDC samples were analyzed by immunohistochemistry for ER, PR, HER2, cytokeratin 5/6 and epidermal growth factor receptor
Two types of mechanisms drive the invasive progression of DCIS: genetic and/or epigenetic modifications occurring in tumor epithelial cells, or nongenetic aberrations as a result of the bidirectional interactions between cancer epithelial cells and their microenvironment. No molecular markers have been convincingly validated to predict which subsets of DCIS are expected to progress to IDC. Nevertheless, a plethora of studies have demonstrated that the most significant changes in gene expression profiles are observed during the transition from normal tissue to DCIS [43] and revealed that the genetic patterns observed in IDC are already present in DCIS, suggesting a probable common origin [24, 44].
\nIn addition to HER2, ER and PR status, similarities between DCIS and IDC begin broadly with chromosomal aberrations that are present in both invasive and in situ stages of disease [45–48]. Several studies have demonstrated that the majority of DCIS cases harbor large copy number alterations that appear to have origins in the transition from normal mammary epithelium to ADH, indicative of a role in early tumorigenesis [40, 48, 49]. The general profile and distribution of DCIS copy number variation have proven to very nearly match that which was previously established for invasive breast cancer with 63% of the peak regions overlapping and 21% within 10 Mb of peak regions established in IDC [46, 48, 50, 51]. Many of the most frequently observed and well‐characterized alterations in invasive cancer, including gains on 1q, 5p, 8q, 12q, 16p, 20q and Xq along with losses on 8p, 9p, 11q, 13q, 14q, 16q, 17p and Xp, were similar among ADH, DCIS and IDC [45, 52]. It has been demonstrated that, on average, 83% (range 59–100%) of matched DCIS and IDC genome sample displays the exact same copy number status [48, 53]. A large body of work has failed to find significant, nonrandom discrepancies in the quantity or quality of DCIS copy number aberrations relative to invasive carcinoma [24, 25, 44, 52, 54–60]. These data support the notion of clonal disease origins and add evidence to the case that DCIS is a precursor lesion to invasive stages. Some studies have shown these genetic alterations to increase in frequency during progression from ADH through DCIS to IDC [24, 45, 49]. This suggests that at least some of the genomic alterations may be required for progression though they could be indicative of global genomic instability resulting in the accumulation of chromosomal gains and losses over time. Some studies utilizing matched synchronous ipsilateral DCIS and IDC have identified potential copy number variations that may be related to progression including amplification of genes mediating proliferative, invasive and migratory function such as the growth factor receptor, fibroblast growth factor receptor 1 [61], V‐myc avian myelocytomatosis viral oncogene homolog (MYC) [62] and cyclin D1 [63] in invasive disease relative to in situ lesions. Despite these findings, it appears that these changes alone are not necessary or sufficient to drive the acquisition of an invasive phenotype and tend to be inconsistent across individual studies [45–47].
In a 2015 study by Abba et al., the full exome, transcriptome and methylome of 30 pure high‐grade DCIS cases were examined. 100% of DCIS cases displayed numerous somatic mutations, 62% harboring mutations in known and potential cancer driver genes; though moderately lower than invasive disease, overall the mutational profile of DCIS is remarkably similar to later stages [40]. Not all DCIS cases with chromosomal copy number variation also exhibit mutations in driver genes, which could suggest that chromosomal alteration proceeds some mutation and is a very early event in the natural history of breast cancer [25, 40]. In fact, only 10% of cases in this study displayed unaltered chromosomal copy number and cancer driver genes. They report evidence of P53 pathway inactivation in every lesion analyzed; regardless of its mutation status or intrinsic subtype, a provocative finding as P53 inactivation is not commonplace in all varieties of invasive disease [64, 65]. Other studies have found P53 mutation in anywhere from 15 to 22% of cases [66–69]. A higher frequency was observed in high‐grade lesions, a trend that is reflected in invasive breast cancer [66, 67]. Overall, this study concluded that on a whole, the molecular profiles they identified in DCIS were indistinguishable from invasive cancer suggesting that the known major genomic anomalies present in later stages of disease are present in their in situ origins. The results of this study corroborate earlier studies comparing the molecular profiles of ADH, DCIS and IDC, all of which find that the stages of breast cancer progression are extremely similar to one another, providing evidence in support of the now widely held notion that the gene and protein expression changes present in IDC proceed the transition from in situ [39, 40, 43, 70–75].
\nA recent study by Lesurf et al. compared DCIS and IDC after stratifying by intrinsic subtype and was able to identify differential and highly specific gene sets that distinguish IDC from DCIS [38]. The gene sets are remarkably distinct with no single gene present in every subtype\'s “invasion signature.” It is thus possible that previous studies lack of stratification by subtype could have generated systematic errors in attempts to identify genetic predictors of progression. Strengthening this case, when genetic analysis is performed without intrinsic subtype stratification, it generates relatively inconclusive differentially expressed gene lists with significant overlap relative to previous lists generated without stratification [39, 43, 70, 74, 76, 77]. These previously generated gene lists have not be overwhelmingly useful as predictive tools for progression of DCIS, making Lesurf\'s findings of invasive signatures valuable. Further work needs to be done to validate these signatures before anyone can comfortably rely on them for consideration in therapy.
\nDCIS has also been analyzed at the proteomic level, and compared to normal ductal and lobular units, this has revealed that alterations in protein expression occur during carcinogenesis [72]. Proteins that have been identified to be significantly differentially expressed between normal structures and in situ carcinoma have functions ranging from control of cytoskeletal architecture, intracellular trafficking, apoptosis, chaperone functions to regulation of genomic stability [72]. Differential expression of actin‐binding proteins was considered to be an unusual finding as remodeling of the actin cytoskeleton tends to be related to lamellar protrusions utilized in invasion and motility, and DCIS is defined as a pre‐invasive lesion [72]. It seems that, in a manner similar to the genomic and transcriptomic variation observed in breast cancer progression, proteomic alterations are an early event in the natural history of tumor progression and do not display extensive changes in the transition from in situ.
Based on the overall lack of differences observed at both the global genomic and transcriptomic levels, some have postulated that epigenetic alterations, inheritable changes that do not modify DNA sequences, may be involved in the transition from in situ to IDC. DNA methylation is a common mechanism of gene promoter silencing [78–80] and has been demonstrated to increase in breast tumorigenesis [81–83]. In comparing normal breast epithelium, ADH, DCIS and IDC, Park et al. noted an increase in methylation status of interrogated breast cancer‐specific CpG islands in the transition from normal to ADH and again when comparing DCIS to ADH though DCIS and IDC did not differ in methylation levels or frequencies. Even the earliest morphologically identifiable stages of breast disease, columnar cell lesions, display an increase in the number of methylated genes, with a similar profile to DCIS and IDC [84]. This suggests that changes in methylation frequency and patterning are an early event in the natural history of breast cancer and may not significantly contribute to the transition between disease stages.
\nOutside of methylation status, chromatin remodeling via modification of histone residues results in differential gene transcription and has been linked to carcinogenesis [85–87]. Hints at the importance of chromatin remodeling in DCIS formation and progression have been demonstrated with over expression of chromatin remodeling proteins associated with transformation of premalignant lesions and poor prognosis in invasive disease [88, 89]. One study has even demonstrated that overexpression of the chromatin remodeling protein EZH2 is able to drive the acquisition of malignant phenotypes in immortalized mammary epithelial cells [88]. Chromatin remodeling also appears to be involved in the epithelial to mesenchymal transition, a process reported to be important in the DCIS transition to IDC [76, 90].
\nAnother mechanism of epigenetic gene regulation is the expression of microRNAs (miRNAs) which are short noncoding sequences that are able to bind and repress translation of messenger RNAs [91, 92]. When compared to normal breast epithelium, DCIS miRNA profiles do display differences such as increased miR‐21 and decreased miR‐98 and let‐7, though these changes are consistent between DCIS and IDC [93]. Some studies have identified potential miRNA invasive signatures highlighting differential expression of a subset of miRNAs in the transition from DCIS to IDC while others have found essentially no difference between the two [38, 40, 94]. Further studies will need to be done to validate the potential pro‐invasive effect of miRNAs that could be involved in the transition to invasive carcinoma.
\nA more recently recognized mechanism of epigenetic regulation is the alternative splicing of mRNA transcripts, allowing for the generation of multiple unique proteins from the same message, which may have differential and even opposing functions [95, 96]. For example, our lab has studied the role of the nuclear co‐activator and oncogene, amplified in breast cancer 1 (AIB1), and we have identified an alternatively processed transcript of the mRNA which generates a shorter form of the protein named AIB1‐Δ4 [97]. We have demonstrated an upregulation of this variant in breast cancer and have associated the alternative processing with loss of a regulatory domain, potentiating the oncogenic function of AIB1 [97]. Our lab has also shown that AIB1 is upregulated in the transition from normal breast to DCIS and maintained in the transition to invasive carcinoma [98]. It is possible that given the enhanced oncogenic activity of Δ4 and correlation of this variant with metastatic capability that alternative splicing of this oncogene could play a role in progression from DCIS to invasive disease. Future investigation into alternative splicing in the acquisition of invasive capacity could yield fruitful results in understanding, predicting and potentially preventing progression.
\nOverall the epigenetic changes interrogated as a potential drivers of tumor progression have returned similar results to genetic and proteomic investigations—differences between invasive and in situ disease are minimal suggesting that epigenetic alterations seen in advanced disease are present from an early stage in the natural progression of breast cancer [81, 84, 99].
Breast cancer cells are integrated within a complex microenvironment that has been increasingly recognized to influence tumor initiation and invasiveness [100, 101]. The tumor microenvironment (TME) is composed of multiple stromal cells (e.g., myoepithelial cells, fibroblasts, immune cells and adipocytes), insoluble extracellular matrix (ECM), newly formed vasculature, as well as growth factors and cytokines [101].
\nThe disruption of the myoepithelial cell layer that separates in situ lesions from the surrounding breast stroma is considered to be the initial step required for DCIS to progress to IDC [102–106]. Normal myoepithelial cells secrete proteinase inhibitors along with factors like thrombospondin, laminin and the oxytocin receptor that ensure the maintenance of BM integrity and suppress epithelial cell proliferation and invasion [104, 105]. By contrast, cancer‐associated myoepithelial cells (CAMs) aid in BM destruction through proteinase production [5, 107]. Though they appear genomically normal, CAMs are significantly different from those associated with normal ductal structures in terms of gene expression and tumor‐suppressive function [73, 102, 106, 108] and engage in paracrine signaling with adjacent cancer cells [109, 110]. One such signaling axis is the upregulation of the chemokine CXCL14 which has been demonstrated to positively influence proliferation, migration and invasion of cancer epithelial cells [73]. Loss of tumor‐suppressive signaling and the loss of this physical cell barrier along with associated ECM signaling unleash progressive potential [5, 102, 106, 111, 112].
In response to impairment of the BM, tumor cells express chemokines (e.g., colony‐stimulating factor 1 receptor) that attract macrophages within the TME [113, 114]. Tumor‐associated macrophages (TAMs), mainly of M2 phenotype, can constitute up to 50% of the breast tumor mass [115], and increased TAMs density has been shown to relate poor prognosis in most human tumors [114, 116]. M2‐type TAMs are critical modulators that potentiate the invasion of tumor cells through various mechanisms: secretion of chemotactic factors (e.g., EGF) [117], pro‐angiogenic molecules (e.g., vascular endothelial growth factor) [118] and anti‐inflammatory cytokines (such as interleukin‐10) [119, 120], as well as remodeling of the ECM [121]. Tumor infiltrating lymphocytes (TILs) have also been identified as a prognostic factor in breast cancer, generally associated with improved survival, decreased distant recurrence and increased metastatic latency predicting a better response to therapeutic interventions and overall survival [122–126]. Though there have been many studies on the importance of immune presence and regulation in advanced breast cancers, the immune infiltrate in DCIS specifically is less well characterized and has only recently started to be evaluated. A novel 2016 study by Thompson et al. investigated the immune microenvironment of 27 DCIS cases of known intrinsic subtype [127, 128]. CD3+ T cells were the predominate lymphocyte subtype across all DCIS cases with CD4+ T‐helper cells making up a slightly larger proportion compared to CD8+ effector T cells. Also present, though at a lower frequency were CD20+ B cells and FoxP3+ T regulatory cells. Interestingly, it was noted that the DCIS cases included that had concurrent invasive disease tended to have more CD20+ B cell and CD8+ T cell infiltrate. Additionally, the DCIS cases known to recur later had greater CD8+ T cells than other subsets of DCIS cases and also displayed an increased relative presence of regulatory T cells than those that did not [127]. These findings suggest that an active adaptive immune response is mounted early in the natural history of breast cancer and that suppression of the host immune system constitutes another crucial step in the malignant progression through the inhibition of immune effector cells (e.g., myeloid‐derived suppressor cells) and the stimulation of immunosuppressive cells (e.g., regulatory T cells) [129].
Cancer‐associated fibroblasts (CAFs) are predominant components of the TME that enhance tumor growth and invasiveness by conferring a mesenchymal‐like phenotype in premalignant mammary epithelial cells [130]. CAFs create a pro‐tumorigenic environment through high deposition, cross‐linking and remodeling of the ECM [131], and by regulating the immune polarization [52]. In breast cancer carcinoma, transforming growth factor beta (TGF‐β), stromal cell‐derived factor‐1, platelet‐derived growth factor α/β and interleukin 6 are the major tumor‐derived factors that have been described to induce CAFs activation [132–134]. Reciprocally, CAFs secrete tumor‐promoting factors, such as hepatocyte growth factor, that stimulate the invasive behavior of DCIS cells [135]. Co‐implantation of CAFs with DCIS cells has been shown to increase the invasive capacity of the in situ lesions [136, 137]. For instance, the presence of CAFs resulted in activation of cyclooxygenase‐2 (COX‐2) in the epithelial component, driving cancer progression [136]. It should be noted that COX‐2 expression was demonstrated to be one of three markers, along with P16 and Ki‐67 that were found to be associated with significantly increased risk of invasive recurrence within 8 years of initial diagnosis and treatment of DCIS [138].
Factors that mediate ECM remodeling and degradation have been of interest in studying the transition of DCIS to invasive disease as destruction of the BM is a hallmark of progression. Several studies have shown matrix metalloproteinases (MMPs) such as MMP1, 2 11, 12 and 13 as well as other proteases and protease inhibitors such as cathepsins, PLAU, SERPINS and metallopeptidase inhibitors to be regulated, up and down respectively, in both DCIS and invasive cancer‐associated stromal cells [73, 74, 139]. These expression changes are further linked to poor prognosis and likely related to the acquisition of invasive capacity [73, 74, 139]. Lyons et al. suggested that mammary gland involution, which is a natural driving force of ECM remodeling following pregnancy [140–142], may recapitulate alterations that occur in the initiation of tumor progression [143]. They demonstrated in a mouse model of involution that xenografted MCF10DCIS.com cells grown in this environment formed larger more invasive lesions marked by increased fibrillar collagen deposition and COX‐2 expression and that anti‐inflammatory treatments with NSAIDs were able to at least partially prevent this progression [143].
In order to sustain their expanding neoplastic growth and eventually disseminate to distant sites, tumors are capable of stimulating the formation of new blood vessels, a process referred to as angiogenesis [144]. Strikingly, the tumor‐associated neovasculature is observed early during carcinogenesis in both murine and human premalignant, noninvasive lesions [145, 146]. The transition from dormant nonvascularized hyperplasia to vascularized proliferative tumor requires the cooperation of various TME cell types (e.g., endothelial and pericytes) and is regulated by counteracting molecules, of which the main pro‐ and anti‐angiogenic factors are vascular endothelial growth factor‐A and thrombospondin‐1, respectively [147].
Our understanding of the natural history of early‐stage breast cancer remains challenging due to the tumor heterogeneity and requires the implementation of experimental models that are capable of mimicking all aspects of the disease. Cell lines and mouse models are valuable tools routinely used to investigate the mechanisms underlying the initiation and progression of breast cancer and will be reviewed in this section.
\nMost in vitro studies aiming to model early‐stage breast cancer are based on the utilization of immortalized mammary epithelial cells
\nMCF10A cell line is the most commonly used breast epithelial cell line to model normal breast epithelium. This immortal cell line was generated from the fibrocystic breast tissue of a 36‐year‐old patient and emerged spontaneously as a result of continuous trypsin‐versene passages [150]. These cells are considered as “normal” breast epithelial cells based on various characteristics commonly found in the normal glandular epithelium, including lack of tumorigenicity, anchorage‐dependent growth, as well as hormonal and growth factor‐dependent proliferation in vitro. MCF10A cells are ER negative and express wild‐type P53 [151] along with markers of basal‐like cells, such as P63 [152, 153]. Although MCF10A cells are non‐transformed and exhibit near diploidy, cytogenetic analyses revealed that these cells are karyotypically abnormal following immortalization. Their genetic abnormalities include amplification of the oncogene MYC and the deletion of the chromosomal locus containing genes regulating the cellular senescence, especially P14ARF and P16 [150]. These latter molecular characteristics render MCF10A cell line particularly adapted for oncogenic transformations. Cui et colleagues recently reported that MCF10A cells do not fully recapitulate in vitro the architectural features of normal human breast tissue most likely due to epigenetic derivations driven by the immortalization process and a continuous culture [154].
In vitro systems using primary human mammary epithelial cells (HMEC) are believed to be a more reliable model of normal breast epithelial cells. Usually easily isolated from reduction mammoplasty tissues, the life span and propagation of HMEC in vitro remain challenging as they stop doubling and undergo cellular senescence after several passages [155]. In addition, these cells tend to lose their lineage commitment as well as their capacity to grow and normally differentiate when cultured ex vivo. To overcome the senescence block, primary human epithelial cells have been immortalized using exogenous expression of viral oncogenes [156] and the telomerase reverse transcriptase [157]. None of these strategies is capable of maintaining these cells in culture without permanently altering their normal phenotype and genetic background. Schlegel and colleagues in collaboration with our laboratory recently established a novel method that can be used to indefinitely propagate a wide range of normal primary epithelial cells, including breast cells [158, 159]. This technique is based on the coculture of primary epithelial cells in presence of irradiated fibroblasts and requires the utilization of a specialized medium containing a Rho‐kinase inhibitor. The resultant cells, also referred to as conditionally reprogrammed cells (CRCs), although highly proliferative, remain karyotypically normal, non‐tumorigenic [158] and exhibit hallmarks of adult stem cells [159, 160]. Because human breast CRCs can be genetically modified in culture and implanted into mouse models as discussed below, the CRC system appears as an in vitro method of choice to study the phenotypic and molecular alterations underlying the benign to malignant transition in breast cancer.
To explore the mechanisms promoting the invasive progression of DCIS, the scientific community has at its disposal few cellular models, although no single cell line is capable of fully recapitulating the different subtypes of DCIS tumors.
\nThe majority of research studies focused on early‐stage breast cancer utilized the premalignant MCF10A series established by Miller and Colleagues [161, 162]. One of these variants, termed MCF10DCIS.com, was isolated upon successive passages in culture of lesions obtained from xenografted MCF10AT cells [162]. At the molecular level, MCF10DCIS.com is a ER‐negative basal‐like cell line that expresses high levels of signaling proteins well‐known to play a crucial role in malignant progression, including CD44v, HER2, COX‐2, Smad4, Stat3, Pak4 and the phosphorylated forms of ERK and AKT [163]. Similarly, a gain of function mutation conferring an increased oncogenic potential to the phosphatidylinositol 3‐kinase has also been found in MCF10DCIS.com cells [164]. Of note, although MCF10DCIS.com cells are considered as a model of early‐stage disease, these cells secrete a significant amount of the metastatic galectin‐3‐binding protein [165], which suggests that they also contain precursors with metastatic capacities. The essential advantage of using MCF10DCIS.com cells relies on their ability to give rise to fast‐growing and comedo‐like DCIS tumors when injected into xenograft mouse model [162, 166]. The particular features of the tumors derived from MCF10DCIS.com xenograft will be described in the next section.
Eleven breast cancer cell lines, referred to as SUM, have been generated by Forozan et al. from different subtypes of primary breast tumors [167]. Two of them, called SUM‐102 and SUM‐225 cells, were immortalized from human DCIS tumors containing microinvasive lesions or from recurrent lesions formed in the chest wall of a patient with DCIS history that did not receive chemotherapy treatment, respectively. SUM‐102 cells express CK19 and are considered as basal B‐type breast cancer cells [167, 168]. These cells also overexpress Cyclin D1 while they possess mutations in PIK3CA, P16 [169] and checkpoint kinase 2 genes [170]. On the other hand, SUM‐225 cells are ER and PR negative, whereas they are amplified for HER2 and are thus classified as luminal epithelial cells [167, 171]. Of note, a P53 missense mutation frequently observed in breast cancer within the sequence encoding the DNA‐binding region has also been found in SUM‐225 cells [171]. Like MCF10DCIS.com cells, SUM‐225 cells generate tumors resembling human DCIS lesions when injected into immuno‐compromised mice [166].
Band and colleagues developed another series, named 21T, including four cell lines established from the tumor tissues of a 36‐year‐old woman that was first diagnosed with stage 3 intraductal carcinoma, then developed lung metastases 1 year later [172]. 21PT and 21NT cells were both isolated and immortalized from the primary breast tumor and were found to resemble ADH and DCIS, respectively. Phenotypically, 21PT cells are normal spindle epithelial cells, whereas 21NT cells are polygonal‐shaped tumor cells of different sizes. At the molecular level, these two cell lines are aneuploid, HER2‐amplified and are believed to not express ER and PR, reflecting the original patient biopsy. The most striking difference between 21PT and 21NT cells relies on their ability to form tumors when grown into immunodeficient mice, and this tumorigenic property was shown to be restricted to 21NT cells [172, 173].
To date, two additional early‐stage breast cancer cell lines have been reported: h.DCIS.01 cells established from columnar cell hyperplastic lesions [77] and FSK‐H7 cells isolated from human DCIS tumors positive for HER2 [166]. Similar to the previous cell lines, h.DCIS.01 and FSK‐H7 cells are capable of producing xenograft tumors in vivo as reviewed previously [77, 166]. In addition to these established cell lines, various technologies have been developed to generate primary breast cancer cell lines representing the full spectrum of human breast cancer subtypes, such as the mammary‐optimized EpiCult‐B technology and the CRC system (described above) [158, 159, 174]. The CRC system is particularly advantageous as it allows for isolation and rapid expansion of tumor cells from a core needle biopsy of human or murine breast cancer [158, 174]. Notably, primary murine CRCs are able to form tumors recapitulating the original carcinoma when implanted orthotopically into syngeneic mice [159].
\nAs previously discussed in this chapter, signals from the breast microenvironment play a key role in the differentiation and maintenance of normal breast epithelial cells [8], as well as during breast cancer initiation and progression [101]. For these reasons, homotypic culture of breast cancer cell lines does not provide the optimal system for studying the multicellular complexity of breast carcinogenesis. This latter limitation emphasizes the importance of developing in vitro culture systems that allow investigations of the cross talk between breast cancer epithelial cells and the surrounding stroma.
Breast cancer cells cultured ex vivo in three‐dimensional and heterotypic systems represent advanced and effective tools for elucidating the morphological and molecular changes governing the epithelial‐stromal interactions during breast cancer invasive progression. Besides recapitulating the breast cellular complexity, organotypic 3D cultures are also practicable systems for experimental research and manipulations of cell lines.
\nMorphogenesis and homeostasis of the normal breast epithelium depend on the balanced relationship between ductal epithelial cells and the ECM [175]. Conversely, the altered communication between epithelial cells and ECM results in the loss of polarity together with the invasion of epithelial cells through the ECM and ultimately contributes to cancer initiation and progression. Breast epithelial cells grown in three‐dimensional (3D) systems can recapitulate the architectural and functional features of the glandular epithelium in vivo in response to molecular signals provided by the ECM substratum [176, 177]. The development of biologically relevant 3D models relies on matching specific matrices and culture media with specific cell types. Particularly, mammary epithelial cells are capable of forming differentiated and functional organoids that resemble normal mammary acini when grown within substrata rich in collagen I [178] or laminin [179], and under culture conditions allowing their survival and proliferation [180, 181]. The vast majority of the 3D cultures of breast epithelial cells imply the utilization of an ECM secreted by the Englebreth‐Holm‐Swarm mouse tumor cells and commercially available as Matrigel™ [179]. Matrigel™ is composed of laminin, collagen IV, entactin and proteoglycans and is supposed to mimic the ductal BM. Monotypic and Matrigel™‐based 3D culture assays recapitulating the breast epithelial signaling have been originally developed by Brugge, Bissell and colleagues in order to unravel the morphogenetic processes supporting the normal mammary gland development and its tumorigenesis [180–182]. Notably, Petersen et al. reported that normal breast cells seeded singly within Matrigel™ are capable of forming spherical, polarized and growth‐arrested acini‐like structures with a central hollow lumen and deposition of BM rich in laminin V and collagen IV [176, 180]. By contrast, breast tumor cells failed to adopt acini‐like phenotypes and instead evolved into poorly differentiated, nonpolarized and highly disordered aggregates when grown in Matrigel™ [176].
\n3D culture systems offer the opportunity to investigate a large spectrum of phenotypic effects mediated by oncogenes and tumor suppressors on the architecture of breast epithelia by using two main strategies. The first strategy intends to reconstruct the tumorigenic phenotypes as a result of targeted genetic modifications in normal epithelial cells. For example, Muthuswamy et al. demonstrated that MCF10A cells genetically engineered to overexpress the oncogene HER2 can give rise to hyperproliferative multi‐acinar structures showing filled lumina and loss of the apicobasal polarization when cultured in homotypic 3D assays [183]. These disorganized structures are characterized by the absence of invasive properties and thus mirror the histopathological hallmarks of precancerous epithelial cells, especially human DCIS. The second strategy aims to genetically manipulate breast cancer cells to possibly restore the organized and polarized phenotype observed in the normal breast duct. Recently, our laboratory followed this strategy to delineate the functional role of the oncogene AIB1 during DCIS progression [98]. We demonstrated that small hairpin RNA (shRNA)‐mediated knockdown of AIB1 in MCF10DCIS.com cells generates more normal acini‐like spheroids with deposition of laminin V to the periphery when these knockdown cells are grown in Matrigel™, overall suggesting that AIB1 plays a key role in the maintenance of the DCIS‐like structure in 3D culture.
\nBesides alterations of the normal breast acinar architecture, the invasion of malignant epithelial cells into the surrounding stroma relies on their migratory properties and implies the protease‐mediated disruption of the BM barrier [73]. 3D culture techniques can be applied to assess the migratory behavior of breast cancer cells through specific 3D matrices in response to particular cytokines [184]. As an illustration, Zaman and colleagues revealed that the co‐overexpression of the oncogenes HER2 and 14‐3‐3ζ in MCF10A cells significantly induces their motility within 3D type I collagen matrices in a stiffness‐dependent manner [185].
\nHomotypic organoid models involving a single cell type grown within reconstituted BM matrices remain the most simplistic approach used to appreciate the epithelial‐stromal interactions. Significant technological progresses had been made in the past decade to provide the cancer cell scientists with a variety of evolved 3D coculture systems reflecting more faithfully the breast cellular complexity in vivo.
Multiple organotypic coculture systems have been elaborated to selectively investigate the cross talk between preneoplastic breast epithelial cells and particular stromal cells, such as myoepithelial cells, fibroblasts and macrophages.
As previously discussed, myoepithelial cells play an essential role in preventing breast cancer dissemination by expressing genes specifically responsible of maintaining a polarized bilayered acinar organization [186]. To decipher the molecular mechanisms underlying the tumor‐suppressive role of myoepithelial cells, Petersen and colleagues recently developed 3D cocultures of human primary luminal and myoepithelial cells isolated from either normal breast reduction mammoplasty or breast tumor tissues [5]. They confirmed that luminal epithelial cells embedded in type I collagen matrices required the presence of normal myoepithelial cells to form polarized acini‐like structures. By contrast, 3D collagen cocultures of luminal epithelial cells with tumor‐derived myoepithelial cells fail to generate properly polarized organoids as a consequence of decreased synthesis of functional laminin I by tumor myoepithelial cells.
In addition to being the most abundant cancer‐associated stromal cells present in the TME, CAFs have been largely shown to modulate the invasive transition of breast cancer [130]. To better comprehend the influence of the cross talk fibroblasts‐epithelial cells during early‐stage breast cancer, Sadlonova et al. grew premalignant breast MCF10AT cells on top of Matrigel™ in the presence of primary fibroblasts [187]. Only fibroblasts purified from normal breast reduction mammoplasty were able to notably suppress MCF10AT cell growth in 3D culture, whereas breast cancer‐derived fibroblasts reduced the proliferation of these transformed cells to a lesser extent. Those results further indicate that upon their malignant conversion, breast cancer epithelial cells become insensitive to the tumor growth inhibitors synthesized by stromal fibroblasts, which underlines the importance of establishing new treatment paradigms for early‐stage breast cancer based on recovering the tumor‐suppressive function of fibroblasts.
Similar heterotypic 3D coculture strategies have been applied to elucidate the molecular mechanisms by which macrophages modulate the behavior of tumor cells during breast carcinogenesis [188, 189]. As an example, Balkwill and colleagues plated MCF‐7 cells or human mammary epithelial cells genetically immortalized with hTERT, on top of Matrigel™ [188]. Then, they tested the invasive phenotype of these two breast cancer cells along with the level of expression of 22 inflammatory‐related genes in the presence of macrophages, previously isolated from human bone marrow and seeded into a modified Boyden chamber to avoid direct cell‐to‐cell contact. This method permitted them to prove that the cancer cell ability to invade through Matrigel™ is induced when cocultured with macrophages. This invasive phenotype was further correlated to increased activation of JNK and NF‐Kappa B pathways. Linde et al. investigated the macrophage‐mediated invasive properties of tumor cells using an approach that integrates macrophages into cocultures of tumor cells with fibroblasts [189]. They plated tumor cells on top of collagen I‐rich dermal equivalents containing macrophages derived from bone marrow alone or together with primary dermal fibroblasts. Using this tri‐culture model, they concluded that activation of macrophages toward M2 phenotype promotes cancer cell invasion through the proteolytic degradation of the BM likely due to increased levels of MMP‐2 and MMP‐9 [189]. Although this model was developed using tumor cells derived from squamous cell carcinoma, it was successfully generated in both a murine and human background, and its applicability can presumably be extended to study the interactions between early‐stage breast tumor cells and TAMs.
\nThe establishment of a wide collection of heterotypic 3D models and their interchangeable utilization have allowed the extensive study of the molecular roles of each cellular component within the breast TME during carcinogenesis. In addition, these models are easily reproducible and particularly handy for the development of targeted therapies. However, even if organotypic 3D cultures are preferred to in vitro models for the identification of new stromal targets for breast cancer progression, only in vivo models have yet been able to thoroughly recapitulate the complexity of the tumor‐stromal interactions that govern breast tumor initiation and invasiveness.
Xenografts of human breast cancer cell lines or tissues implanted into immuno‐compromised recipient mice represent powerful tools for understanding the multiple aspects of the human disease within an in vivo context. Athymic nude, severe combined immune deficient (SCID), NOD/SCID IL2Rgammanull (NGS) and “humanized” NSG mice are the most commonly utilized immunodeficient animals [190]. Two types of xenograft mouse models of breast cancer are usually established: through subcutaneous injection [162], or after orthotopic transplantation of cancer cells or tissues into the mammary fat pad or the duct [166, 173, 191]. Breast cancer cell lines can be genetically manipulated prior to being grafted into mice, which allows to study the tumorigenic properties of specific factors on the tumor take, growth and dissemination in vivo [98]. On the other hand, the actual human cancer tissue can be used to rapidly generate xenograft tumors recapitulating the cellular and molecular heterogeneity inherent to a particular cancer [191]. As a result, xenograft mouse models are frequently employed to predict the tumor response to clinically relevant drugs as well as their possible adverse effects and thus have served as preclinical models for multiple clinical trials [190]. In order to identify the possible factors driving the invasive transition of DCIS, various xenograft models of human DCIS have also been developed based on the utilization of the DCIS cell lines reviewed above, especially MCF10ADCIS.com [162, 166, 192], 21NTci [173] and SUM‐225 cells [166].
\nAs previously discussed
As detailed previously, 21PT and NT cell lines are primary tumor‐derived cells believed to mimic early‐stage breast cancer when implanted into the mammary fat pad of female nude mice at 8–9 weeks of age [173]. Souter et al. reported that 21PT cells were not tumorigenic or metastatic in vivo and instead formed xenograft structures that shared features of ADH. Furthermore, atypical/neoplastic and normal/benign epithelial cells were shown to coexist within 21PT xenograft tissues. Conversely, 21NT cells were able to give rise to malignant lesions in about 20% of the mice, and the resultant tumors displayed intermediate to high‐grade DCIS lesions, with no evidence of invasive progression. Interestingly, 21NT‐derived lesions exhibited phenotypic traits identical to the ones obtained in MCF10ADCIS.com xenograft mouse model, although in contrast to 21NT tumors, MCF10ADCIS.com tumors ultimately progress to IDC as a result of RAS‐induced transformation.
Most of the xenograft models are obtained upon subcutaneous injection of cancer cells and thus fail to replicate the early steps of breast carcinogenesis that occur inside the mammary duct. In order to better recreate the natural progression of DCIS within conditions mimicking the stromal microenvironment, Medina and colleagues developed a novel method based on the intraductal transplantation of human breast cancer cells into immunodeficient female mice [166]. To generate this unique mouse intraductal model, they injected the previously described DCIS cell lines, MCF10ADCIS.com, SUM‐225 and FSK‐H7 cells, through the nipple directly into the mammary ducts of 6‐ to 10‐week‐old SCID‐beige mice. Following intraductal transplantation, all three cells lines were able to form DCIS‐like lesions that slowly evolved into IDC in 90% of the mice. Unlike subcutaneous injection, the intraductal transplantation of MCF10ADCIS.com cells into immuno‐compromised mice gave rise to cribriform DCIS, whereas SUM‐225 and FSK‐H7 cells displayed comedo and apocrine‐like DCIS lesions, respectively. Using immunostaining analyses, Behdoh et al. further reported that SUM‐225 and FSK‐H7 xenograft samples are characterized by the overexpression of HER2 along with moderate expression of CK8 and 19. MCF10ADCIS.com‐derived tumors were also positively stained for CK8 although they contained lesions classified as basal‐like as they express CK5. More recently, Behbod et colleagues extended the intraductal mouse model to primary human DCIS [191]. They successfully xenotransplanted DCIS cell lines, derived from eight different patient samples, within the mammary duct of 8‐ to 10‐week‐old virgin NSG mice. At 8 weeks, the tissues collected from the xenografted tumors exhibited noninvasive lesions that closely resemble human DCIS and that retain the histopathological and molecular hallmarks of the patient\'s original biopsy. Strikingly, the engraftment of primary DCIS cells obtained from human tumors was only permitted using mice depleted in both mature T and B cells, suggesting that T and B cells both regulate the tumor growth of primary cells into in vivo models
As the fundamental role exerted by the tumor microenvironment during early‐stage breast cancer is increasingly recognized, the subcutaneous implantation of cancer cells into mouse models deficient in T and B lymphocytes remains problematic and may result in decreased tumor take and abnormal cancer progression. To overcome this critical limitation, “humanized” mouse models have been generated based on the engraftment of human CD34+ hematopoietic stem cells onto irradiated NOD/SCID mice [193] and the orthotopic implantation into cleared mammary fat pads humanized using stromal cells of human origin [194]. Humanized mice represent very promising models for better defining the tumorigenic properties of tumor‐associated stromal cells during breast cancer development. This strategy can further be used to create a wide range of analogous models reflecting the various subtypes of DCIS.
Genetically engineered mouse models (GEMM) constitute invaluable resources for experimentally examining the in vivo function of specific oncogenes or tumor suppressor genes within immunocompetent animals. The primary benefit of employing GEMM as model of human cancer is that this approach allows modulation of the expression of particular genes in a tissue and time‐specific fashion. Many GEMM models of breast cancer are currently available and are of two kinds: transgenic models in which specific oncogenes are overexpressed, and knockout models created upon targeted deletion of tumor suppressors [195]. In the following section, we will focus on GEMM that were found to develop tumors containing early‐stage breast cancer lesions, especially DCIS.
\nIn transgenic mouse models, the transcription of particular oncogene is induced throughout the mammary luminal epithelium under the control of strong mammary‐specific epithelial promoters, of which the mouse mammary tumor virus (MMTV) and the whey acidic protein (WAP) are the most frequently utilized [195]. Transgenic mammary carcinoma models usually give rise to highly penetrant tumors after short latency. To date, few transgenic mouse models of human breast cancer have been shown to develop premalignant and DCIS‐like lesions. They are discussed below.
Given the undeniable association of
The WAP promoter sequence has also been employed to induce mammary intraepithelial neoplasia in mice. Tzen et al. initially applied this strategy to drive the murine mammary epithelial cell transformation through WAP‐mediated increased transcription of the SV40 large T antigen (SV40 TAg) [201]. SV40 TAg possesses the unique properties to physically interact with both P53 [202] and the retinoblastoma protein [203] and abrogate their tumor‐suppressive function, leading to cellular hyperproliferation. Deppert and colleagues recently expanded the histological analysis of the WAP‐T transgenic mouse model and reported that these animals carried multifocal DCIS‐like carcinomas progressing to IDC after a short latency period [204]. Notably, the resultant in situ carcinoma was composed of differentiated lesions showing tubular and papillary architecture comparable to those diagnosed in human DCIS.
Because MMTV and WAP were shown to be regulated by hormones [205, 206], Dr. Green\'s laboratory engineered a novel transgenic model by transcriptionally overexpressing the SV40 TAg in the mammary and prostate glands under the hormone‐independent control of the C3(1) component of the prostate steroid‐binding protein (PSBP) 5′ flanking sequence [207]. All C3(1)‐SV40 TAg female mice bore mammary tumors that were found to share similar histologic and molecular hallmarks of human infiltrating ductal carcinoma in a predictable time‐dependent manner. Eight‐week‐old transgenic mice, in fact, did exhibit ADH‐like lesions that evolve into DCIS at about 12 weeks, invasive carcinoma at 16 weeks of age and ultimately metastasized into the lung with a 15% incidence. In addition, well‐known chromosomal and molecular aberrations driving mammary carcinogenesis were also observed in C3(1)‐SV40 TAg mice, thus suggesting that this transgenic model provides the unique opportunity to assess the antitumor potential of targeted therapies.
Increased levels of ERα in mammary epithelial cells are believed to be correlated to initiation and progression of premalignant breast cancer [208]. In order to corroborate this hypothesis, our collaborator Dr. Furth established a unique mouse model with dominant gain of ERα by breeding together three types of transgenic animals expressing: (i) the tetracycline‐responsive transactivator tTA under control of MMTV promoter (MMTV‐tTA); (ii) SV40 TAg under control of the tetracycline‐responsive promoter (tetop); (iii) FLAG‐tagged ERα under control of tetop [209, 210]. Strikingly, 4‐month‐old triple transgenic mice (MMTV‐tTA/tetop‐SV40 TAg/tetop‐ERα) gave rise to ERα positive and estrogen‐sensitive mammary tumors that contained preneoplastic lesions identical to those found in human ADH and DCIS. Using the double transgenic mice MMTV‐tTA/tetop ERα, Frech et al. further reported that the rate of mammary epithelial cell proliferation was higher in ADH and DCIS lesions and was accompanied by increased expression of cyclin D1 in the nuclear compartment of these cells [210]. Altogether, this work reported for the first time that high ERα expression can promote the benign to cancerous transformation of mammary epithelial cells in vivo and provided unique mouse models to unravel the neoplastic events regulated by ERα signalings.
Due to the time‐consuming nature of using GEMM and their costs, various methods have recently emerged based on the establishment of stable mammary intraepithelial neoplasia outgrowth (MIN‐O) lines derived from tumors carried by GEMM, such as MMTV‐PyV‐mT [211]
In summary, the molecular and cellular profiling of early‐stage breast cancer using in vitro and in vivo models that reflect the heterogeneity of the disease have drastically allowed for improved understanding of the mechanisms underlying invasive progression of DCIS. As no individual model is capable of recapitulating the complexity of the human tumors, efforts should be made in the future to develop integrated strategies for better defining the drivers of early‐stage progression of breast cancer and thus open new avenues for targeted therapy.
This work was funded in part by RO1CA205632 (ATR) and the Nina Hyde Center Breast Cancer Fund (ATR).
Hepatocellular carcinoma (HCC) is the most common primary malignant tumor of the liver in adults, the fifth most common cancer in the world and also the third most common cancer of cancer-related deaths [1]. It is the malignancy of hepatocytes with varying degrees of differentiation [2]. The most common cause of death in patients with HCC is cirrhosis. Despite all the unknowns, heptocarcinogenesis is a multistep process, and chronic inflammation plays the major role [2, 3, 4, 5, 6].
HCC has a multifactorial etiology, and its incidence and prevalence varies by country [7]. Although the incidence of HCC is different in different geographies, the incidence increases with age [8]. It is more common in men than in women (male:female ratio; ranging between 2:1 and 4:1 in various countries). Cirrhosis, viral hepatitis, alcohol, aflatoxin, metabolic diseases, metabolic syndrome characterized by fatty liver are the main causes of HCC etiology. Although the activation of the WNT/B-catenin pathway is one of the main events in HCC, the effects of viral antigens on the nucleus, mutations, and DNA instability constitute the pathogenesis of HCC [9]. There are also molecular studies showing that activation of the JAK/STAT pathway also contributes to the development of SH-HCC [1, 10].
Most patients with HCC have underlying liver cirrhosis. Cirrhosis is therefore considered a major risk factor for HCC [8, 11, 12]. Although macronodular cirrhosis is considered as a higher risk for HCC than micronodular cirrhosis, cirrhotic liver can create HCC for any reason [13]. Cirrhosis also has a geographical distribution, the etiology of cirrhosis is chronic viral hepatitis in Asian countries, and nonviral causes in European and American countries [14]. Despite this known association between cirrhosis and HCC, HCC also develops from the noncirrhotic liver [15, 16, 17].
Most HCCs develop from the background of chronic viral hepatitis, including hepatitis B and Hepatitis C [18, 19, 20, 21]. Viral hepatitis-related HCCs are more common in countries where hepatitis B and hepatitis C are more prevalent, such as Asia and Africa. Viral-related HCC appears to be decreasing in countries where clinical follow-up increases, and whom include hepatitis B vaccine in regular vaccination programme. Integration of hepatitis B virus into the host hepatocyte genome is thought to initiate hepatocarcinogenesis. In the etiology of HCC, hepatitis C is as important as hepatitis B [22, 23, 24]. Being men and older, having coinfection (such as HBV, HIV), alcohol use, diabetes, and fatty liver constitute a high risk for HCC formation. Even the development of HCC in liver coinfected with hepatitis C and hepatitis B viruses, is higher than in those infected with other viruses [13]. It is thought that ongoing liver damage and accompanying regeneration caused by the immune response and direct cytopathic effect in hepatitis C infection induce malignant transformation [25, 26].
Consumption of foods contaminated with aflatoxins produced by fungi can lead to the formation of HCC [8, 27, 28]. Aflatoxin B1, one of the toxin types, is thought to be mainly responsible for HCC formation [8]. It contributes to the formation of HCC by making mutations (Guanin and Thymine mutations) in DNA via cytochrome p450. Aflatoxin exposure is thought to affect patients with chronic HCV hepatitis more [8, 29, 30].
The relationship between alcohol use and HCC is both by direct effect and being a cofactor in viral infections [31, 32]. Reactive oxygen radicals, that occur while alcohol is metabolized to acetaldehyde, initiate hepatocarcinogenesis by causing damage and transformation in DNA. HCC development in alcoholic cirrhosis is in the form of DNA instability caused by DNA hypomethylation [33, 34, 35, 36].
HCC can develop in some of the livers with metabolic diseases. However, the development of HCC is more common with hereditary hemochromatosis, tyrosinemia and α1-antiripsin deficiency [37, 38, 39, 40, 41]. In these diseases, the direct toxic effect of accumulations (such as iron), mutation (p53 mutation), immunological abnormalities and DNA damage by lipid peroxidation initiate the development of HCC [8, 42, 43].
Metabolic syndrome is a mortal endocrinopathy that is accompanied by systemic disorders such as abdominal obesity that begins with insulin resistance, diabetes, dyslipidemia, hypertension and coronary artery disease. This situation has led to an increase in HCC formation, which has the characteristics of metabolic syndrome [44, 45, 46, 47]. The risk of HCC increases 2–3 times in patients with diabetes [37, 48, 49]. The increase in metabolic syndrome in developed countries also brought an increase in nonalcholic fatty liver disease (NAFLD) [50, 51, 52, 53, 54, 55].
In obese patients, the decrease in the release of fatty acids from adipose tissue, tumor necrosis factor-α and adiponectin causes insulin resistance and thus chronic hyperinsulinemia. Insulin and insulin growth factor-1 (IGF-1) contribute to hepatocarcinogenesis by preventing apoptosis and increasing cellular proliferation with the signals they send to insulin receptors and IGF-1 receptors [8].
Since steatohepatitic HCC (SH-HCC) will be mentioned here, NAFLD, steatohepatitis and their associated HCC formation mechanism are explained in a little more detail.
There are many studies on the incidence and prevalence of HCC in NAFLD cases, with rates varying between 3 and 35% [51, 56, 57]. Steatohepatitis varies between 3 and 5%. In some cohort studies, the rate of development of HCC (1-year cumulative incidence) was reported as 2–5% in patients with NAFLD compared to hepatitis C cases. The 5-year incidence was reported as 11% [51, 58]. In another study, the annual cumulative rate was 2–6%. In a retrospective study, NAFLD was detected in 21.2% of HCC cases. In fact, 23% of NAFLD patients without histopathologically and radiologically significant cirrhosis developed HCC [59]. In a different study, HCC develops in 5% of patients with cirrhosis secondary to NAFLD [53]. In cohort studies with large case series, both steatosis and steatohepatitis in nontumoral liver were found to be statistically significant with HCC. Moreover, a close relationship between the steatohepatitic variant of HCC (SH-HCC), which has been recently defined, and NAFLD has been described and demonstrated [22, 53, 58, 60]. Although its relationship with fatty liver diseases has been clarified, there are studies showing that SH-HCC can also develop in viral hepatitis [16, 61].
The clinical manifestations of HCC are quite ambiguous and are related to the tumor and underlying chronic liver disease [1]. Usually, patients show signs in advanced stages and even miss the chance of treatment. Patients may present with upper abdominal pain, hepatomegaly, splenomegaly, weight loss, jaundice or decompensated liver finding such as ascites [1, 8]. HCC most commonly spreads intrahepaticly via the portal vein [1]. While HCC spreads with intrahepatic portal vein branches, the main portal vein and hepatic vein involvement can also be seen. Invasion of the bile duct causes liver decompensation, resulting in rapid ascites accumulation, obstructive jaundice, variceal hemorrhages, and hepatic encephalopathy [8]. Although extrahepatic dissemination is rare, it can metastasize to the lung, lymph nodes, bone, and adrenal gland in advanced disease [1]. Paraneoplastic syndrome findings such as hypoglycemia, hypercholesterolemia, hyperkalemia, gynecomastia, carcinoid syndrome, hypertrophic pulmonary osteoarthropathy, osteopetrosis, hypertension, hyperthyroidism, porphyria cutanea tarda can be seen [8]. Median suvival in patients with clinical findings who have the chance for curative treatment is around 1–3 months, and survival over 1 year is also unusual. Today, thanks to definitive treatments and advanced surgeries, patients at risk of developing HCC are followed more closely and the tumor is diagnosed at an early stage [8, 28]. Radiologic imaging methods (ultrasound, computed tomography, magnetic resonance imaging, angiography) are used for the diagnosis of liver masses and HCC [8, 17, 62, 63].
HCC is a highly heterogeneous tumor. Heterogeneity is both molecular and morphological [64, 65]. Understanding the heterogeneity is important for the diagnosis, treatment and follow-up of the disease [64].
HCCs below 2 cm are called small HCC (s-HCC) and early HCC (e-HCC) [1, 64, 66]. These tumors are divided into two as prominent nodular or indistinct nodules [64]. Early-HCC is in the form of nodules with indistinct borders and usually develops from a dysplastic nodule background. They are well differentiated, develop from the background of fibrosis-cirrhosis, and are radiologically hypovascular and rarely vascular invasion (5%) [1, 64]. Small-HCC has a prominent pseudocapsule, is well-moderately differentiated, radiologically hypervascular, and invades more frequently (40%) [1, 64]. Pedinculated HCC has a growth pattern protruding from the capsular surface [67]. Diffuse HCC is in the form of proliferation of small tumor nodules and resembles cirrhotic nodules (cirrhotomimetic) [64]. SH-HCC is more solid than other HCC subtypes and has more golden-yellow color due to the lipid contains. When the macroscopic specimen is carefully examined, fibrotic bands that divide the tumor into lobules can be seen. The tumor usually tends to be well-circumscribed or nodular and may range in diameter from 0.5 cm to 11 cm [68]. The prognosis of SH-HCC is similar to that of classical HCC [40, 57, 69, 70]. Although nontumoral liver can be cirrhotic or noncirrhotic, it is usually yellowish-brown in color suggestive of fatty liver (Figure 1).
Tumor with nodular border is seen in the brown-yellow noncirrhotic liver. The tumor is lobulated by fibrotic bands and has yellow areas.
After these macroscopic definitions and macroscopic heterogeneity, it is necessary to mention microscopic heterogeneity. This heterogeneity is also reflected in the histopathological subtyping of HCC [71]. In the 5th edition of WHO classification of the tumors of the digestive system (2019), the subtypes of HCC are as follows; fibrolamellar, scirrhus, cear cell type, steatohepatitic, macrotrabecular massive, chromophobe, neutrophil-rich, lympocyte-rich [1]. More on SH-HCC will be mentioned here. SH-HCC is a newly identified subtype of HCC. It accounts for approximately 5–20% of all HCCs [1]. It is characterized by steatohepatitic features such as steatosis in tumor cells, balloon degeneration, inflammation, Mallory-Denk bodies and pericellular fibrosis [58, 72]. Tumor is usually related to MetS and steatohepatitis is detected in the background. [22, 39, 57, 61, 69, 72]. Some studies have shown that steatosis and interstitial fibrosis are the main findings for SH-HCC [22, 40]. However, the minimum amount of steatosis in the steatohepatitic area in some tumors that are histomorphologically SH-HCC, the presence of only steatosis in some cases, the presence of steatotic areas or cells in HCC are confusing points in the diagnosis of SH-HCC. Despite all this confusion, the steatohepatitic area in HCC is diagnostic for SH-HCC. For the histopathological diagnosis of SH-HCC, the cut-off for steatohepatitic features was described more than 5% of the tumor before but later moved to 50% [10, 39, 64, 72]. Hepatocellular carcinoma morphologically has 4 histological growth patterns: trabecular, solid (compact), pseudoaglandular (pseudoacinar), and macrotrabecular (trabecular thickness consisting of more than 10 cells) [1]. When SH-HCC is examined microscopically, a steatotic tumor is seen, separated from the generally steatotic liver (cirrhotic or non-cirrhotic) by a nodular or infiltrative margin. Large fat droplets are detected in tumor cells. Mallory-Denk bodies are detected in most tumors. Thin connective tissue growth (pericellular fibrosis), trabecular fibrosis, and randomly distributed collagen bundles surrounding tumor cells can be easily selected. Trabecular fibrosis, including randomly distributed collagen bundles in the tumor, and fibrosis surrounding tumor cells (pericellular) can be easily distinguished. Inflammation in the tumor is also remarkable. The inflammation is lymphocyte predominant with sparse plasma cells. More prominent neutrophil and lymphocyte infiltrations can be detected around tumor cells which contains Mallory-Denk bodies. The nuclei of tumor cells have atypia. This atypia is mild in well-differentiated tumors and quite pronounced in poorly differentiated tumors. They may even have bizarre nuclei suggested of sarcomas or pleomorphic carcinomas. However, mitotic activity is very low. Again, as in classical HCC and other subtypes, the tumor does not contain portal tracts and unpaired arteries can be seen (Figures 2–4) [1, 10, 45, 68, 72, 73]. The differentiation of SH-HCC is the same as that of classical HCC and is graded as well differentiated (Grade1: Tumor cells resemble mature hepatocytes with minimal to mild atypia), moderately differentiated (Grade 2: Distinctly malignant and histomorphology strongly suggests hepatocellolar differentiation) and poorly differentiated (Grade 3: Clearly malignant, but histomorphology strongly suggests spectrum of poorly differentiated carcinomas) [1]. Most SH-HCCs are moderately differentiated and have a trabecular pattern and a pseudoglandular pattern [52].
The tumor (pale area) is located in the center of the figure, surrounded by cirrhotic nodules (a, Hematoxylin and Eosin-H&E). Masson’s trichrome (b) and reticulin (c) stains, both the tumor and its surrounding micronodular cirrhotic background are more prominent.
The parenchymal invasion area of steatohepatitic HCC is seen (arrows) (a), the tumor is seen adjacent to the fatty cirrhotic nodule (stars) (b), presence of large lipid droplets and chronic inflammation (arrows) (c), Masson’s trichrome stain shows thick fibrous septa (arrows) (d).
Dense inflammation and fibrosis (a), pleomorphism (b), Mallory-Denk bodies (arrows) (c), and ballooning (cells with pale cytoplasm) (d) are seen in different areas of the tumor.
Immunohistochemical antibodies are helpful and supportive in the diagnosis of HCC [74]. Although heppar-1, glypican-3, glutamine synthetase, arginase, heat shock protein-70 (HSP-70), β-catenin and sinusoidal staining with CD34, canalicular staining pattern with polyclonal carcinoma embryogenic antigen (pCEA) and CD10 antibodies are used in the diagnosis of HCC, immune studies for SH-HCC are limited (Figure 5) [22, 68, 72, 75].
Glutamine synthetase shows positive cytoplasmic staining (a), CD10 antibody shows positive canalicular staining (b).
The histopathological diagnosis of SH-HCC is usually easy in cases with explant and resection. However, tru-cut biopsies, which represent a small part of the tumor, may have diagnostic difficulties. These diagnostic difficulties are due to both the heterogeneity of the tumor and its similar morphological appearance to NAFLD with advanced fibrosis. A tru-cut biopsy from focal nodular hyperplasia (FNH) with fatty changes sometimes can be confused with a diagnosis of nodular and well differentiated SH-HCC. This difference between the diagnosis in the tru-cut biopsy and the resection material should not be interpreted as a misdiagnosis. Before interpreting it as an erroneous diagnosis, it should be remembered that this diagnostic difference is due to the heterogeneous and fat-containing nature of the tumor. Pathologists should remember that bile duct proliferation, presence of central scar (histologically and radiologically), and thick-walled abnormal vascular structures in the fibrous septa are more common in FNH when examining this tru-cut biopsy. Since fibrosis can be seen in both SH-HCC and FNH, it may not clarify the differential diagnosis. Non-invasive border and immunohistochemical staining (sinusoidal CD34 staining, glypican-3 positivity and diffuse glutamine synthetase staining) may be helpful in the differential diagnosis of steatohepatitis [8, 11, 68, 72]. Differentiation from classical HCC can be made by evaluating morphological and immune markers together [68]. In spite of all this, it would be appropriate to consult a pathologist experienced in liver pathology in cases where tumor specification could not be made.
The relationship between NAFLD, NASH, and HCC (especially SH-HCC) is now known. Adequate tumor sampling should be performed in resection materials, explants, particularly when identifying subtypes of large-diameter HCCs. It should be noted that classical HCC and other subtypes, including SH-HCC, have a heterogeneous histomorphology. While patients with metabolic syndrome, insulin resistance, obesity, fatty liver and steatohepatitis are followed up, careful radiological examination should be performed for SH-HCC that may develop from this background. In other words, the terminology of “neoplastic steatogenesis” should be kept in mind.
The Internet has irrevocably changed the dynamics of scholarly communication and publishing. Consequently, we find it necessary to indicate, unambiguously, our definition of what we consider to be a published scientific work.
",metaTitle:"Prior Publication Policy",metaDescription:"Prior Publication Policy",metaKeywords:null,canonicalURL:"/page/prior-publication-policy",contentRaw:'[{"type":"htmlEditorComponent","content":"A significant number of working papers, early drafts, and similar work in progress are openly shared online between members of the scientific community. It has become common to announce one’s own research on a personal website or a blog to gather comments and suggestions from other researchers. Such works and online postings are, indeed, published in the sense that they are made publicly available. However, this does not mean that if submitted for publication by IntechOpen they are not original works. We differentiate between reviewed and non-reviewed works when determining whether a work is original and has been published in a scholarly sense or not.
\\n\\nThe significance of Peer Review cannot be overstated when it comes to defining, in our terms, what constitutes a published scientific work. Peer Review is widely considered to be the cornerstone of modern publishing processes and the key value-adding contribution to a scholarly manuscript that a publisher can make.
\\n\\nOther than the issue of originality, research misconduct is another major issue that all publishers have to address. IntechOpen’s Retraction & Correction Policy and various publication ethics guidelines identify both redundant publication and (self)plagiarism to fall within the definition of research misconduct, thus constituting grounds for rejection or the issue of a Retraction if the work has already been published.
\\n\\nIn order to facilitate the tracking of a manuscript’s publishing history and its development from its earliest draft to the manuscript submitted, we encourage Authors to disclose any instances of a manuscript’s prior publication, whether it be through a conference presentation, a newspaper article, a working paper publicly available in a repository or a blog post.
\\n\\nA note to the Academic Editor containing detailed information about a submitted manuscript’s previous public availability is the preferred means of reporting prior publication. This helps us determine if there are any earlier versions of a manuscript that should be disclosed to our readers or if any of those earlier versions should be cited and listed in a manuscript’s references.
\\n\\nSome basic information about the editorial treatment of different varieties of prior publication is laid out below:
\\n\\n1. CONFERENCE PAPERS & PRESENTATIONS
\\n\\nGiven that conference papers and presentations generally pass through some sort of peer or editorial review, we consider them to be published in the accepted scholarly sense, particularly if they are published as a part of conference proceedings.
\\n\\nAll submitted manuscripts originating from a previously published conference paper must contain at least 50% of new original content to be accepted for review and considered for publication.
\\n\\nAuthors are required to report any links their manuscript might have with their earlier conference papers and presentations in a note to the Academic Editor, as well as in the manuscript itself. Additionally, Authors should obtain any necessary permissions from the publisher of their conference paper if copyright transfer occurred during the publishing process. Failure to do so may prevent Us from publishing an otherwise worthy work.
\\n\\n2. NEWSPAPER & MAGAZINE ARTICLES
\\n\\nNewspaper and magazine articles usually do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense. Articles appearing in newspapers and magazines rarely possess the depth and structure characteristic of scholarly articles.
\\n\\nSubmitted manuscripts stemming from a previous newspaper or magazine article will be accepted for review and considered for publication. However, Authors are strongly advised to report any such publication in an accompanying note to the External Editor.
\\n\\nAs with the conference papers and presentations, Authors should obtain any necessary permissions from the newspaper or magazine that published the work, and indicate that they have done so in a note to the External Editor.
\\n\\n3. GREY LITERATURE
\\n\\nWhite papers, working papers, technical reports and all other forms of papers which fall within the scope of the ‘Luxembourg definition’ of grey literature do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense.
\\n\\nAlthough such papers are regularly made publicly available via personal websites and institutional repositories, their general purpose is to gather comments and feedback from Authors’ colleagues in order to further improve a manuscript intended for future publication.
\\n\\nWhen submitting their work, Authors are required to disclose the existence of any publicly available earlier drafts in a note to the Academic Editor. In cases where earlier drafts of the submitted version of the manuscript are publicly available, any overlap between the versions will generally not be considered an instance of self-plagiarism.
\\n\\n4. SOCIAL MEDIA, BLOG & MESSAGE BOARD POSTINGS
\\n\\nWe feel that social media, blogs and message boards are generally used with the same intention as grey literature, to formulate ideas for a manuscript and gather early feedback from like-minded researchers in order to improve a particular piece of work before submitting it for publication. Therefore, we do not consider such internet postings to be publication in the scholarly sense.
\\n\\nNevertheless, Authors are encouraged to disclose the existence of any internet postings in which they outline and describe their research or posted passages of their manuscripts in a note to the Academic Editor. Please note that we will not strictly enforce this request in the same way that we would instructions we consider to be part of our conditions of acceptance for publication. We understand that it may be difficult to keep track of all one’s internet postings in which the researcher´s current work might be mentioned.
\\n\\nIn cases where there is any overlap between the Author´s submitted manuscript and related internet postings, we will generally not consider it to be an instance of self-plagiarism. This also holds true for any co-Author as well.
\\n\\nFor more information on this policy please contact permissions@intechopen.com.
\\n\\nPolicy last updated: 2017-03-20
\\n"}]'},components:[{type:"htmlEditorComponent",content:'A significant number of working papers, early drafts, and similar work in progress are openly shared online between members of the scientific community. It has become common to announce one’s own research on a personal website or a blog to gather comments and suggestions from other researchers. Such works and online postings are, indeed, published in the sense that they are made publicly available. However, this does not mean that if submitted for publication by IntechOpen they are not original works. We differentiate between reviewed and non-reviewed works when determining whether a work is original and has been published in a scholarly sense or not.
\n\nThe significance of Peer Review cannot be overstated when it comes to defining, in our terms, what constitutes a published scientific work. Peer Review is widely considered to be the cornerstone of modern publishing processes and the key value-adding contribution to a scholarly manuscript that a publisher can make.
\n\nOther than the issue of originality, research misconduct is another major issue that all publishers have to address. IntechOpen’s Retraction & Correction Policy and various publication ethics guidelines identify both redundant publication and (self)plagiarism to fall within the definition of research misconduct, thus constituting grounds for rejection or the issue of a Retraction if the work has already been published.
\n\nIn order to facilitate the tracking of a manuscript’s publishing history and its development from its earliest draft to the manuscript submitted, we encourage Authors to disclose any instances of a manuscript’s prior publication, whether it be through a conference presentation, a newspaper article, a working paper publicly available in a repository or a blog post.
\n\nA note to the Academic Editor containing detailed information about a submitted manuscript’s previous public availability is the preferred means of reporting prior publication. This helps us determine if there are any earlier versions of a manuscript that should be disclosed to our readers or if any of those earlier versions should be cited and listed in a manuscript’s references.
\n\nSome basic information about the editorial treatment of different varieties of prior publication is laid out below:
\n\n1. CONFERENCE PAPERS & PRESENTATIONS
\n\nGiven that conference papers and presentations generally pass through some sort of peer or editorial review, we consider them to be published in the accepted scholarly sense, particularly if they are published as a part of conference proceedings.
\n\nAll submitted manuscripts originating from a previously published conference paper must contain at least 50% of new original content to be accepted for review and considered for publication.
\n\nAuthors are required to report any links their manuscript might have with their earlier conference papers and presentations in a note to the Academic Editor, as well as in the manuscript itself. Additionally, Authors should obtain any necessary permissions from the publisher of their conference paper if copyright transfer occurred during the publishing process. Failure to do so may prevent Us from publishing an otherwise worthy work.
\n\n2. NEWSPAPER & MAGAZINE ARTICLES
\n\nNewspaper and magazine articles usually do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense. Articles appearing in newspapers and magazines rarely possess the depth and structure characteristic of scholarly articles.
\n\nSubmitted manuscripts stemming from a previous newspaper or magazine article will be accepted for review and considered for publication. However, Authors are strongly advised to report any such publication in an accompanying note to the External Editor.
\n\nAs with the conference papers and presentations, Authors should obtain any necessary permissions from the newspaper or magazine that published the work, and indicate that they have done so in a note to the External Editor.
\n\n3. GREY LITERATURE
\n\nWhite papers, working papers, technical reports and all other forms of papers which fall within the scope of the ‘Luxembourg definition’ of grey literature do not pass through any extensive peer or editorial review and we do not consider them to be published in the scholarly sense.
\n\nAlthough such papers are regularly made publicly available via personal websites and institutional repositories, their general purpose is to gather comments and feedback from Authors’ colleagues in order to further improve a manuscript intended for future publication.
\n\nWhen submitting their work, Authors are required to disclose the existence of any publicly available earlier drafts in a note to the Academic Editor. In cases where earlier drafts of the submitted version of the manuscript are publicly available, any overlap between the versions will generally not be considered an instance of self-plagiarism.
\n\n4. SOCIAL MEDIA, BLOG & MESSAGE BOARD POSTINGS
\n\nWe feel that social media, blogs and message boards are generally used with the same intention as grey literature, to formulate ideas for a manuscript and gather early feedback from like-minded researchers in order to improve a particular piece of work before submitting it for publication. Therefore, we do not consider such internet postings to be publication in the scholarly sense.
\n\nNevertheless, Authors are encouraged to disclose the existence of any internet postings in which they outline and describe their research or posted passages of their manuscripts in a note to the Academic Editor. Please note that we will not strictly enforce this request in the same way that we would instructions we consider to be part of our conditions of acceptance for publication. We understand that it may be difficult to keep track of all one’s internet postings in which the researcher´s current work might be mentioned.
\n\nIn cases where there is any overlap between the Author´s submitted manuscript and related internet postings, we will generally not consider it to be an instance of self-plagiarism. This also holds true for any co-Author as well.
\n\nFor more information on this policy please contact permissions@intechopen.com.
\n\nPolicy last updated: 2017-03-20
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After that, sections about methods of production and structures of nanocomposites will be detailed. Finally, some properties and potential applications that have been achieved in polymer nanocomposites will be highlighted.",book:{id:"6854",slug:"nanocomposites-recent-evolutions",title:"Nanocomposites",fullTitle:"Nanocomposites - Recent Evolutions"},signatures:"Amanda Dantas de Oliveira and Cesar Augusto Gonçalves Beatrice",authors:[{id:"249768",title:"Ph.D.",name:"Amanda",middleName:null,surname:"Oliveira",slug:"amanda-oliveira",fullName:"Amanda Oliveira"},{id:"254512",title:"Ph.D.",name:"Cesar",middleName:"Augusto Gonçalves",surname:"Beatrice",slug:"cesar-beatrice",fullName:"Cesar Beatrice"}]},{id:"57267",title:"Natural Fibers for Sustainable Bio-Composites",slug:"natural-fibers-for-sustainable-bio-composites",totalDownloads:2373,totalCrossrefCites:15,totalDimensionsCites:23,abstract:"Over the past decade, the concept of utilizing green materials has become more mainstream. With considerable awareness of preserving the environment, sincere efforts across the globe can be cited in looking for bio-degradable and bio-based sources. Applications of bio-based materials from renewable and bio-degradable sources for preparation of higher valued green chemicals and bio-based products have forced many scientists to investigate the potential use of natural fibers as reinforcement materials for green bio-composites. Cellulosic fibers are becoming very interesting for bio-based material development as they possess advantages with their mechanical properties, low density, environmental benefits, renewability, and economic feasibility. Recently, natural-fiber polymer composites have received much attention for different industrial applications because of their low density and renewability. The bio-composites with natural fiber components are derivatives of depleting resources and can be considered to have substantial environmental and economic benefits. This chapter addresses the potential utilization of natural fiber for the development of green polymer composite materials, with the objective to elucidate the possibility of using these bio-based materials for various industrial applications.",book:{id:"6233",slug:"natural-and-artificial-fiber-reinforced-composites-as-renewable-sources",title:"Natural and Artificial Fiber-Reinforced Composites as Renewable Sources",fullTitle:"Natural and Artificial Fiber-Reinforced Composites as Renewable Sources"},signatures:"Tri-Dung Ngo",authors:[{id:"208798",title:"Ph.D.",name:"Tri-Dung",middleName:null,surname:"Ngo",slug:"tri-dung-ngo",fullName:"Tri-Dung Ngo"}]},{id:"57169",title:"Development of Hemp Fibers: The Key Components of Hemp Plastic Composites",slug:"development-of-hemp-fibers-the-key-components-of-hemp-plastic-composites",totalDownloads:1823,totalCrossrefCites:4,totalDimensionsCites:6,abstract:"Plant fibers in general and hemp fibers in particular have great prospects for their use in various innovative applications such as ecological, biodegradable, and renewable resources with unique properties. Such properties together with the increased strength due to high-cellulose content and specific morphological parameters are widely used to produce plant fiber–based plastic composites. The properties of plant fibers that may influence the properties of composites depend on crop processing, but the basis for them is provided during fiber development in planta. It is known that two types of bast fibers are developed in the hemp stem: primary fibers formed from procambium cells and secondary fibers that originate as a result of cambium activity. Both types of fibers may significantly vary in their yield and quality depending on the variety and growth conditions. Differences in the anatomical and morphological characteristics of the two types of hemp fibers, together with peculiarities in the composition and architecture of cell wall, influence the technical parameters of the raw material quality. Based on our study of both primary and secondary fiber development in hemp stem that was focused on the two key stages, intrusive elongation and deposition of thick cell wall layers, we suggest the set of parameters that can influence the quality of the mature fibers and trace their biological origin.",book:{id:"6233",slug:"natural-and-artificial-fiber-reinforced-composites-as-renewable-sources",title:"Natural and Artificial Fiber-Reinforced Composites as Renewable Sources",fullTitle:"Natural and Artificial Fiber-Reinforced Composites as Renewable Sources"},signatures:"Chernova Tatyana, Mikshina Polina, Salnikov Vadim, Ageeva\nMarina, Ibragimova Nadezda, Sautkina Olga and Gorshkova\nTatyana",authors:[{id:"158372",title:"Dr.",name:"Tatyana",middleName:null,surname:"Chernova",slug:"tatyana-chernova",fullName:"Tatyana Chernova"},{id:"209953",title:"Prof.",name:"Tatyana",middleName:null,surname:"Gorshkova",slug:"tatyana-gorshkova",fullName:"Tatyana Gorshkova"},{id:"209955",title:"Dr.",name:"Polina",middleName:null,surname:"Mikshina",slug:"polina-mikshina",fullName:"Polina Mikshina"},{id:"209956",title:"Dr.",name:"Marina",middleName:null,surname:"Ageeva",slug:"marina-ageeva",fullName:"Marina Ageeva"},{id:"209957",title:"MSc.",name:"Olga",middleName:null,surname:"Sautkina",slug:"olga-sautkina",fullName:"Olga Sautkina"}]},{id:"50950",title:"Carbon Nanotube-Based Polymer Composites: Synthesis, Properties and Applications",slug:"carbon-nanotube-based-polymer-composites-synthesis-properties-and-applications",totalDownloads:4770,totalCrossrefCites:39,totalDimensionsCites:77,abstract:"The present chapter covers the designing, development, properties and applications of carbon nanotube-loaded polymer composites. The first section will provide a brief overview of carbon nanotubes (CNTs), their synthesis, properties and functionalization routes. The second section will shed light on the CNT/polymer composites, their types, synthesis routes and characterization. The last section will illustrate the various applications of CNT/polymer composites; important properties, parameters and performance indices backed by comprehensive literature account of the same. The chapter concludes with the current challenges and future aspects.",book:{id:"5167",slug:"carbon-nanotubes-current-progress-of-their-polymer-composites",title:"Carbon Nanotubes",fullTitle:"Carbon Nanotubes - Current Progress of their Polymer Composites"},signatures:"Waseem Khan, Rahul Sharma and Parveen Saini",authors:[{id:"149897",title:"Dr.",name:"Parveen",middleName:null,surname:"Saini",slug:"parveen-saini",fullName:"Parveen Saini"}]},{id:"56947",title:"Waste and Recycled Textiles as Reinforcements of Building Materials",slug:"waste-and-recycled-textiles-as-reinforcements-of-building-materials",totalDownloads:1559,totalCrossrefCites:6,totalDimensionsCites:9,abstract:"Currently, the use of composite materials in the construction areas has had a great impact on the society; mainly, those related with sustainability and environment aspects. Daily proposals aimed at overcoming the properties of traditional materials that arise, which include emergent materials either from waste or recycled products. One of them is related to the textile materials, which include fibers such as wool, hemp, linen, and cotton. In the past decade, special attention has been focused on the used clothes, which represent a source of raw materials environmentally responsible and economically profitable. Textile materials are discarded daily around the world, representing approximately 1.5% of the generated waste. Blue jeans are the most used clothing in the world, and they are elaborated by one of the most commonly used natural textile fibers—cotton. Textile materials have been reused in different applications, for example, in the production of poor-quality wires, crushed to manufacture noise and temperature insulation materials, and as fillers or reinforcements of concrete. In this chapter, different topics are described that include: (a) environmental impact of textile waste—a result of massive consumption of clothing, (b) recycling and reuse of textile waste, and (c) waste and recycled textile materials used as building materials.",book:{id:"6233",slug:"natural-and-artificial-fiber-reinforced-composites-as-renewable-sources",title:"Natural and Artificial Fiber-Reinforced Composites as Renewable Sources",fullTitle:"Natural and Artificial Fiber-Reinforced Composites as Renewable Sources"},signatures:"Patricia Peña Pichardo, Gonzalo Martínez-Barrera, Miguel Martínez-\nLópez, Fernando Ureña-Núñez and Liliana I. 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Dr. Koprowski has authored more than a hundred research papers with dozens in impact factor (IF) journals and has authored or co-authored six books. Additionally, he is the author of several national and international patents in the field of biomedical devices and imaging. 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He completed a one-year Post-Doctoral Fellowship awarded by the DFAIT (Foreign Affairs and International Trade Canada) at the Institute of Biomedical Engineering of the University of New Brunswick (Canada) in 2010. Currently, he is Professor in the Faculty of Electrical Engineering (UFU). He has authored and co-authored more than 200 peer-reviewed publications in Biomedical Engineering. He has been a researcher of The National Council for Scientific and Technological Development (CNPq-Brazil) since 2009. He has served as an ad-hoc consultant for CNPq, CAPES (Coordination for the Improvement of Higher Education Personnel), FINEP (Brazilian Innovation Agency), and other funding bodies on several occasions. He was the Secretary of the Brazilian Society of Biomedical Engineering (SBEB) from 2015 to 2016, President of SBEB (2017-2018) and Vice-President of SBEB (2019-2020). He was the head of the undergraduate program in Biomedical Engineering of the Federal University of Uberlândia (2015 - June/2019) and the head of the Centre for Innovation and Technology Assessment in Health (NIATS/UFU) since 2010. He is the head of the Postgraduate Program in Biomedical Engineering (UFU, July/2019 - to date). He was the secretary of the Parkinson's Disease Association of Uberlândia (2018-2019). Dr. Andrade's primary area of research is focused towards getting information from the neuromuscular system to understand its strategies of organization, adaptation and controlling in the context of motor neuron diseases. 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For 20 years, he has studied the analysis and processing of biomedical images, emphasizing the full automation of measurement for a large inter-individual variability of patients. Dr. Koprowski has authored more than a hundred research papers with dozens in impact factor (IF) journals and has authored or co-authored six books. Additionally, he is the author of several national and international patents in the field of biomedical devices and imaging. Since 2011, he has been a reviewer of grants and projects (including EU projects) in biomedical engineering.",institutionString:null,institution:{name:"University of Silesia",institutionURL:null,country:{name:"Poland"}}}]},{type:"book",id:"7218",title:"OCT",subtitle:"Applications in Ophthalmology",coverURL:"https://cdn.intechopen.com/books/images_new/7218.jpg",slug:"oct-applications-in-ophthalmology",publishedDate:"September 19th 2018",editedByType:"Edited by",bookSignature:"Michele Lanza",hash:"e3a3430cdfd6999caccac933e4613885",volumeInSeries:2,fullTitle:"OCT - Applications in Ophthalmology",editors:[{id:"240088",title:"Prof.",name:"Michele",middleName:null,surname:"Lanza",slug:"michele-lanza",fullName:"Michele Lanza",profilePictureURL:"https://mts.intechopen.com/storage/users/240088/images/system/240088.png",biography:"Michele Lanza is Associate Professor of Ophthalmology at Università della Campania, Luigi Vanvitelli, Napoli, Italy. His fields of interest are anterior segment disease, keratoconus, glaucoma, corneal dystrophies, and cataracts. His research topics include\nintraocular lens power calculation, eye modification induced by refractive surgery, glaucoma progression, and validation of new diagnostic devices in ophthalmology. \nHe has published more than 100 papers in international and Italian scientific journals, more than 60 in journals with impact factors, and chapters in international and Italian books. 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