Wound healing is a complex regulated process that results in skin scar formation in postnatal mammals. Chronic wounds are major medical problems that can confer devastating consequences. Currently, there are no treatments to prevent scarring. In the early fetus wounds heal without scarring and the healing process is characterized by relatively less inflammation compared to adults; therefore, research aimed at reducing the inflammatory process related to wound healing might speed healing and improve the final scar appearance. The skin is the largest organ of the body and acts as the first line of defense against pathogens, toxins, and trauma. It also plays a critical role in fluid homeostasis and provides sensory functions and thermal regulation. Damage or loss of skin integrity resulting from an injury or disease can lead to significant morbidity and even death.Wound healing is a complex, regulated process in which regulated collagen deposition, in response to tissue injury, results in scar formation. Its mechanisms include inflammation, fibroplasia, and scar maturation.Sometimes cutaneous wounds do not progress to normal healing with formation of a final mature scar formation but to a continuing inflammatory process, which can lead to a more aggressive carcinogenic transformation in long time of evolution (Marjolin's Ulcer). Many chronic wounds are the result of chronic inflammation. In contrast to adult wound healing, the early gestation fetus displays a remarkable ability to heal wounds without scarring. Fetal wounds heal rapidly and are characterized by a relative lack of inflammation . The intIn this paper, we review how to curb inflammation in cutaneous wound healing; the following lists the main topics discussed in this paper.Topics about Curbing Inflammation in Wound HealingGeneral TopicsWet environment.Primary closure and sutures.Drugs or supplement:curcumin,protein supplementation,nutraceuticals,neurotensin (NT),photodynamic therapy (PDT),doxycycline,α) antibody.tumour necrosis factor-alpha ,hyperbaric oxygen,silver dressings,negative pressure wound therapy (NPWT).Biomaterials and cellular therapy.Specific WoundsDiabetic wounds.Vascular wounds.Irradiated wounds.Repair of the skin consists of three phases: an initial inflammatory phase and then a proliferative/repair phase and it concludes with a remodeling phase, which results in scar formation in postnatal mammals.In response to tissue injury, inflammatory cells are recruited to wounded tissue. The acute inflammatory response is followed by proliferation of fibroblasts, which are responsible for synthesizing collagen and extracellular matrix. Fibroblasts can differentiate into myofibroblasts, which are responsible for collagen deposition and wound contraction. Ultimately, a scar results from accumulation of extracellular matrix. Despite scar remodeling during maturation, normal architecture is never completely restored ; only 70The early stage of inflammation is regarded as a critical period of the wound healing process, essential for clearing contaminating bacteria and creating an environment conducive to the succeeding events of tissue repair and regeneration . The injβ and tumor necrosis factor- (TNF-) α, have been shown to regulate the recruitment and function of neutrophils. In irradiated mice, an investigation of the ability of exogenous IL-1β or transforming growth factor-β to reverse radiation-induced defective wound healing found that IL-1β enhanced wound tensile strength [α is a major cytokine secreted by macrophages and neutrophils during the inflammation phase of wound healing; it is elevated in early wound healing [Cytokines have been widely studied because they are important to wound healing; they regulate the activity of the cells that produce the healing response to tissue injury . Severalstrength . TNF-α i healing .In all phases of wound repair, extracellular matrix (ECM) proteins play a key role in directing the fate and activities of progenitor and reparative cells. Immediately after injury, the ECM orchestrates the recruitment of platelets and directs the inflammatory cell response that initiates the hemostatic and the cellular debridement phases . These cA wet or moist environment in wounds has been shown to promote reepithelialization and result in reduced scar formation more than a dry environment . The infThe modern approach of employing a moist environment for the treatment of wounds was introduced in the early 1960s by Winter , who shoManufacturers responded to Winter's research findings and provided a wide range of moist dressings, such as hydrocolloids that absorb the wound fluid beneath a semiocclusive dressing , foams , 17, algAdult human skin wounds heal with varying degrees of scar formation . ScarrinSurgical sutures are used universally to achieve proper wound approximation by supporting the strain of closure until sufficient healing has occurred. Approximating the wound wedges with sutures decreases the wound area subjected to the healing process and abbreviates the inflammatory phase, which decreases scarring due to the faster pace of healing. All sutures are foreign bodies; therefore their presence triggers a local immune response. Thereby sutures may, paradoxically, potentiate and extend the inflammatory phase of wound healing, resulting in undesirable outcomes, such as wound breakdown, suture “spitting,” hypertrophic scar, or keloid formation. The degree of inflammation induced varies with suture composition, which ranges from relatively inert polypropylene and nylon to highly inflammatory silk and plain gut suture. As a result, numerous investigations have attempted to alter suture composition with the goal of reducing the inflammatory response; however, no specific composition has gained clinical relevance or widespread practice .α, IL-1β, and MMP-9, and increased levels of the anti-inflammatory cytokine IL-10 and antioxidant enzymes, SOD, catalase, and GPx, at the wound site in diabetic rats. This data allows us to suggest that curcumin could be an additional novel therapeutic agent for the management of impaired wound healing in diabetics and radiated tissues [Kant et al.  found th tissues , 32.β, IL-6, and TNF-α) and anti-inflammatory cytokine (IL-4 and IL-10) levels nearly to the levels of control animals. These findings support the hypothesis that delayed wound healing in diabetic animals is caused by deregulation of signaling molecules that are critical mediators of the inflammatory response.Abdel-Salam  demonstrSerra et al.  evaluateBased on findings that peripheral nerves and cutaneous neurobiology contribute to correct wound healing , Moura eReports of animal studies indicate that photodynamic therapy (PDT) improves healing of excisional wounds but the mechanism is uncertain. Mills et al.  first reSerra et al.  documentα activity. In a case series [Therapeutic antibodies such as infliximab can inhibit TNF-e series , inflixiTranscutaneous electrical nerve stimulation (TENS) has been shown to accelerate the healing of chronic wounds in human subjects and induced wounds in animal models , 41. MorHyperbaric oxygen therapy (HBOT), an effective tool that helps skin wound healing, is defined as delivery of 100% oxygen at greater than one atmospheric pressure (ATA) into the core of the wound . Deliver (1)Augmented hydroxylation: it helps improve collagen synthesis. (2)Angiogenesis: HBOT creates a large oxygen gradient between the center and periphery of the wound that strongly stimulates angiogenesis, which is seminal for wound healing . (3)Increased bactericidal activity. (4)Increased collagen deposition.Hyperbaric oxygen works by the following four dominant mechanisms:P = 0.037). This improvement, however, disappeared at the two-week follow-up [Kalani et al. showed that when diabetic foot ulcers were treated with HBOT, after 3 years of therapy, they healed completely in 76% of patients, whereas the ulcers of only 48% patients healed completely with conventional treatment [ollow-up .Ionized silver (Ag+) has both anti-inflammatory and antimicrobial properties with a broad spectrum of action –52. JemeWu et al.  found thAccording to prospective and retrospective clinical and experimental studies , 58, negβ and IL-8 expression in the early phase of inflammation, which may trigger infiltration of neutrophils and thus accelerate bacterial clearance. Furthermore, the success of NPWT of acute wounds can attenuate the expression of TNF-α, and this effect may partly explain why NPWT does not significantly impair wound healing [The impact of NPWT on local expression of proinflammatory cytokines, the number of neutrophils, and the bacterial bioburden living on the wound surface suggest that NPWT of acutely infected soft-tissue wounds leads to increased local IL-1 healing .NPWT not only enhances the granulation tissue in the more superficial layer of the wound bed but also appears to affect deeper layers, with relief of chronic inflammation and tissue stabilization. It is believed that NPWT removes excess fluid and thereby removes proteolytic enzymes that negatively influence the healing process .Interestingly, studies have shown that the activity of some matrix metalloproteinases (MMPs) is elevated in chronic wounds and the presence of these molecules is related to impairing wound healing. For instance, high concentrations of MMP-9 and high MMP-9 : tissue inhibitors of MMP (TIMP-1) ratio in wound fluid predict poor wound healing in diabetic foot ulcers . In addiOn the other hand, data strongly suggest that NPWT influences the microenvironment of the wound by reducing the levels of MMPs and the MMP : TIMP ratio. Mouës et al.  found siIn addition, some studies compared different types of interface material for the application of NPWT with other common dressings. Tuncel et al. , who stuThe use of acellular and cell-based tissue-engineered dermal substitutes has become increasingly routine . Dermal Of note, tissue engineering and regenerative medicine have shown significant promise in treatment of problematic wounds. Initial proof-of-principle studies that investigated the use of exogenously applied adult stem cells for the treatment of chronic wounds provided support for their vulnerary effects , 77. MSCDiabetes is a multisystem disorder, and its complications induce physiological changes in tissues and cells that impair the normal healing process. However, the pathophysiologic relationship between diabetes and impaired healing is complex. Diabetic wounds become halted in the inflammatory phase, which is characterized by a continuing influx of neutrophils that release cytotoxic enzymes, free radicals, and inflammatory mediators that cause extensive collateral damage to surrounding tissue. These destructive processes counteract the healing process in such wounds, and the overproduction of free radicals, which induce oxidative stress, results in detrimental cytotoxic effects that delay wound healing , 80.Increased oxidative stress is one of the most common reasons for the delayed wound healing in the diabetic population . TherefoChronic venous, lower limb ulceration (VLU) affects 1–3% of the adult population worldwide,  and somePathophysiological events involved in the onset of chronic venous ulceration are inflammation, activation of polymorphonucleates (PMNs), and secretion of proteases such as MMPs, which degrade the ECM that supports the vascular and tissue walls. MMPs, neutrophil gelatinase-associated lipocalin (NGAL), and inflammatory cytokines are overexpressed in VLUs, and they could play a central role in pathophysiological mechanisms of skin lesions and delayed wound healing.Approximately 50% of cancer patients receive radiation as an adjuvant therapy. Although radiation therapy technology has progressed substantially in the last decades, patients still suffer from various degrees of nonspecific radiation damage to noncancerous tissues. Radiation ulcers have posed an enormous challenge to reconstructive surgeons; also, they cause great distress to patients, impair their quality of life, and consume high amounts of medical resources. These kinds of chronic wounds can last for several years, and some may even lead to amputations. Strategies for treating these and other chronic wounds may include the use of various growth factors –88, hypeOver the last fifty years, the complex wound healing process became clearer allowing the development of strategies, devices, and medications that help achieve a better final scar, mainly through strategies to curb the wound-related inflammation. Nonetheless, we are still far from definitely solving the chronic wounds or the pathologic scarring issue and, even more, achieving the goal of rapid and scarless healing similar to fetal one.


Macrophage migration inhibitory factor (MIF) and its receptor CD74 appear to be involved in tumorigenesis. We evaluated, by immunohistochemical staining, the tissue expression and distribution of MIF and CD74 in serial sections of human invasive breast cancer tumor specimens. The serum MIF level was also determined in breast cancer patients. We showed a significant increase in serum MIF average levels in breast cancer patients compared to healthy individuals. MIF tissue expression, quantified by a modified Allred score, was strongly increased in carcinoma compared to tumor-free specimens, in the cancer cells and in the peritumoral stroma, with fibroblasts the most intensely stained. We did not find any significant correlation with histoprognostic factors, except for a significant inverse correlation between tumor size and MIF stromal positivity. CD74 staining was heterogeneous and significantly decreased in cancer cells but increased in the surrounding stroma, namely in lymphocytes, macrophages and vessel endothelium. There was no significant variation according to classical histoprognostic factors, except that CD74 stromal expression was significantly correlated with triple-negative receptor (TRN) status and the absence of estrogen receptors. In conclusion, our data support the concept of a functional role of MIF in human breast cancer. In addition to auto- and paracrine effects on cancer cells, MIF could contribute to shape the tumor microenvironment leading to immunomodulation and angiogenesis. Interfering with MIF effects in breast tumors in a therapeutic perspective remains an attractive but complex challenge. Level of co-expression of MIF and CD74 could be a surrogate marker for efficacy of anti-angiogenic drugs, particularly in TRN breast cancer tumor. Breast cancer (BC) is the most frequently occurring malignant disease among women in the Western hemisphere, inflicting one in eight women . DespiteMacrophage migration inhibitory factor (MIF) is a pleiotropic inflammatory cytokine of 12.5-kDa monomeric molecular weight originally described as a T cell lymphokine modulating macrophage motility. Subsequently, MIF was shown to be produced by a variety of immune and non-immune cells such as B- and T-lymphocytes as well as endocrine, endothelial and epithelial cells of diverse histogenetic origin. Pathophysiologically, MIF plays a pivotal role in various autoimmune and inflammatory disorders, such as rheumatoid arthritis, systemic lupus erythematosus, septic shock and atherosclerosis . In addiMIF serum levels are elevated in breast cancer patients  and MIF Looking at a receptor, CD74 is expressed in breast cancer tissue and its presence appears to be correlated with lymph node invasion and triple-negative tumors –19 makinin vitro with some success as an antibody-drug conjugate on solid cancer cell lines positive for CD74  inhibits growth of MIF-expressing breast tumors in mice . Regardifor CD74 .These considerations led us to an immunohistochemical assessment of expression of MIF and CD74 in serial sections of human breast cancer tumor specimens, mapping their profiles in cancer and stromal cells. In parallel, the serum level of MIF was determined in breast cancer patients.Formalin-fixed, paraffin-embedded, residual tissue material of diagnostic biopsies of 96 breast cancer tumors , which wBlood samples from 36 newly diagnosed early breast cancer female patients (BCP) were obtained prospectively for determining serum level of MIF, prior to any breast cancer treatment . Twenty-This study was approved by the Ethics Committee of Erasme Hospital, Brussels, Belgium, according to the international and Belgian laws (P2008/314 and A2013/016).Serum concentration was assayed by a sandwich enzyme-linked immunosorbent assay (ELISA) using a commercial kit . The assays were carried out according to the instructions provided by the supplier. MIF concentrations in serum samples were determined by interpolation from a reference curve established with increasing concentrations of recombinant human MIF.2O2. Sections were counterstained with luxol fast blue prior to light microscopy examination.For immunostaining of MIF, after antigen retrieval by microwave treatment, sections were pretreated with hydrogen peroxide to block endogenous peroxidase activity. Thereafter, they were exposed to casein to avoid false-positive staining. These steps were followed by sequential incubations with (i) primary antibody  poFor immunostaining of CD74 , immunohistochemistry was performed using a procedure similar to that described previously for MIF .MIF and CD74 expressions in the glandular and in the stromal compartments were assessed by light microscopy and quantified according to a modified Allred score .Non parametric analysis was carried out using the Mann-Whitney test and Spearman’s rank order correlations was used.The intensity of immunohistochemical staining for MIF was assessed in tumor-free breast tissues . However, we found a significant inverse correlation between tumor size and stromal positivity  status and the absence of estrogen receptors , including the absence of ER itself. A correlation between CD74 expression in BC, TRN status and lymph node invasion has previously been reported ,18.To date, co-expression of MIF and CD74 has not been studied in breast cancer but it has been described in prostate and non-small cell lung cancer. In prostate cancer, MIF was intense but CD74 staining was weak and patchy . In lungin vitro but causes delayed tumor growth in mice, increasing the prevalence of an immune suppressive myeloid-derived population within the tumor  knockdown by siRNA approach, as reported for clear cell renal carcinoma (Our findings showed an increased extent of MIF expression in cancer cells and in stromal fibroblasts of BC tumor, in contrast to a less uniform increase of CD74 expression mainly in stromal lymphocytes, macrophages and endothelium. This could suggest a pro-oncogenic role of MIF in BC tumors taking place predominantly in the stromal compartment. MIF, secreted by tumor cells, could then modulate the immune microenvironment and its neovascularization, favoring escape from immune surveillance and tumor cell dissemination. Experimental data published show that MIF suppression in breast cancer cell lines does not affect their proliferation he tumor ,36. Xu eC tumors . FinallyC tumors . In vitrarcinoma , shed liIn conclusion, our data support the concept of a functional role of MIF in human breast cancer. In addition to auto- and paracrine effects on cancer cells, MIF could contribute to shape the microenvironment leading to immunomodulation and angiogenesis, these aspects deserving further investigations. Interfering with MIF effects in breast tumors in a therapeutic perspective remains an attractive but complex challenge, notably depending on the development of suitable MIF inhibitors. Level of co-expression of MIF and CD74 could be a surrogate marker for efficacy of anti-angiogenic drugs, particularly in TRN breast cancer tumors.


Tumor cells actively contribute to constructing their own microenvironment during tumorigenesis and tumor progression. The tumor microenvironment contains multiple types of stromal cells that work together with the extracellular matrix and local and systemic factors to coordinately contribute to tumor initiation and progression. Tumor cells and their stromal compartments acquire many genetic and/or epigenetic alternations to facilitate tumor growth and metastasis. The cancer stem cell (CSC) concept has been widely applied to interpreting tumor initiation, growth, metastasis, dormancy and relapse. CSCs have differentiation abilities to generate the original lineage cells that are similar to their normal stem cell counterparts. Interestingly, recent evidence demonstrates that CSCs also have the potential to transdifferentiate into vascular endothelial cells and pericytes, indicating that CSCs can transdifferentiate into other lineage cells for promoting tumor growth and metastasis in some tissue contexts instead of only recruiting stromal cells from local or distant tissues. Although the transdifferentiation of CSCs into tumor stromal cells provides a new dimension that explains tumor heterogeneity, many aspects of CSC transdifferentiation remain elusive. In this review, we summarize the multi-lineage differentiation and transdifferentiation potentials of CSCs as well as discuss their potential contributions to tumor heterogeneity and tumor microenvironment in tumor progression. Cancer stem cells (CSCs), also known as tumor-initiating cells or tumor-propagating cells, refer to a subpopulation of tumor cells that have abilities to self-renew, differentiate and seed new tumors. Accumulating evidence demonstrates that CSCs contribute to tumorigenesis, metastasis, dormancy and relapse –3. CSCs in vivo, indicating that progenitors can convert to leukemia stem cells [et al., reported that MOZ-TIF2, but not BCR-ABL, transforms myeloid progenitors into leukemia initiating cells [et al., discovered that the ectopic expression of transcription factors, such as Twist1 and Snail, or treatment with TGF-β in mammary epithelial cells or cancer cells can induce stem cell-like or cancer stem cell-like phenotypes [in vitro acquire CSC properties, undergoing multi-lineage differentiation and generating hierarchically organized tumors in vivo [in vivo still involves many obstacles. Therefore, CSCs may originate from their normal stem cells, progenitors and/or differentiated somatic cells.Because CSCs exhibit self-renewal and multi-lineage differentiation abilities that are similar to their normal stem cell counterparts, CSCs were initially thought to originate from normal stem cells. Transducing MLL-AF9 fusion protein into myeloid progenitors gives rise to leukemia em cells . Huntly ng cells . All of enotypes . Terminaenotypes . There wenotypes . Skin fi in vivo . Thus, tin vitro and in vivo  in vitro and in vivo [+/EpCAM+ or CD133+ subpopulation initiates tumorigenesis and differentiates into colon cancer cells [According to the CSC theory, CSCs can differentiate into cancer cells and are responsible for tumor growth and metastasis. Dick and colleagues identified a CD34 in vivo . Leukemi in vivo . The difer cells . In brai in vivo . In coloer cells , 10.in vitro and serial transplantation ability in vivo. When treat with the different tumor cell/tissue-derived conditioned medium, the single-cell-cloned CSCs are able to differentiate into respective type of tumor cells [+ subpopulation establishes tumors in vivo with hierarchy cell organization and exhibits MSC plasticity to undergo adipogenic, osteogenic and chondrogenic differentiation in vitro [et al., discovered that Yamoto-synovial sarcoma cells are enriched for CSCs and efficiently drive tumor formation in vivo. Silencing SS18-SSX genes in synovial sarcoma stem cells induces multiple-lineage differentiation in vitro that is similar to MSCs [in vitro and initiate tumorigenesis with the hierarchy organization of cancer cells in vivo.The differentiation of CSCs into cancer cells has been validated in other types of cancers, including pancreatic , prostator cells . In addiin vitro . Naka et to MSCs . CollectCSCs share many properties with normal stem cells, including persistent activation of Notch, Hedgehog, and Wnt pathways, which are involved in the regulation of differentiation and self-renewal of stem cells . Fibrobl+ renal CSCs can generate endothelial cells in vitro and give rise to vessels with a human origin in vivo [et al., further discovered that human breast CSCs undergo endothelial differentiation to generate functional endothelial cells in vitro and in vivo [+ ovarian CSCs can form CD34+ blood vessels in xenograft tumor models. CD44+ ovarian CSCs do not express any endothelial progenitor cell markers, but they can differentiate into endothelial cells in Matrigel. Furthermore, these CD44+ cells form vessel-like structures in a VEGF-independent manner. These three papers provide evidence for the endothelial differentiation of CSCs; however, they lack robust in vivo and clinical evidence that CSCs directly contribute to tumor angiogenesis via transdifferentiation. In 2010, studies on GBM further illustrated that GBM stem cells (GSCs) generate endothelial cells in tumor angiogenesis [et al., reported that a large proportion of the endothelial cells in GBM harbors the same chromosomal alterations as in tumor cells. They isolated CD133+/CD31− cells from GBM samples that differentiate into functional endothelial cells in vitro. This transdifferentiation event was further validated in vivo using a xenogarft model in which human origin of endothelial cells was detected without cell fusion. In addition, selectively targeting GBM stem cell-derived endothelial cells dramatically impairs tumor growth [et al., [+ endothelial cells in GBM samples have GBM genetic mutations, indicating that these endothelial cells are not derived from normal endothelial cells. The purified CD133+/CD144− GBM cells can transdifferentiate into endothelial cells in vitro and in vivo via endothelial progenitor cell status without nuclear fusion. Activated Notch signaling contributes to the transdifferentiation of GSCs into endothelial progenitors [Solid tumors exhibit diverse patterns of neovascularization, including sprouting angiogenesis, vasculogenesis, intussusception, vessel co-option, vascular mimicry and CSC transdifferentiation . Vascula in vivo . Bussola in vivo . The aut in vivo . CD44+ oogenesis , 23. Ricr growth . Wang etgenitors . The autgenitors . These sin vitro and in xenograft model in vivo. This work also took advantage of a lineage-specific fluorescence reporter system to validate that GSCs have the capacity to transdifferentiate into pericytes in vivo. By analyzing patients' GBM samples, the authors found that vast majority of tumor pericytes carry the same genetic alternations as neoplastic cells. Moreover, selectively targeting GSC-derived pericytes disrupts tumor vessels and inhibits tumor growth. Interestingly, GSCs preferentially transdifferentiate into pericytes but not into endothelial cells. Although cells expressing endothelial markers were detected in the cells that were differentiated from GSCs in culture, the numbers were very low [Pericytes attach to endothelial cells to support the neovasculature, maintain the integrity of blood vessels and communicate with each other to regulate endothelial homeostasis , 52. A lvery low . The autvery low . Howeverin vivo, numerous in vitro studies demonstrate that EMT is a well-defined program in carcinoma. By the EMT program, epithelial cells exhibit fibroblast-like morphology and express mesenchymal markers, such as α-SMA, and acquire partial stem cell-like properties. In breast cancer, EMT provides a nonmalignant stroma. The α-SMA-expressing cells that have originated from epithelial tumor cells promote tumor growth in nude mice [in vivo. Recent studies reveal that constitutively activating EMT suppresses the stemness of human epithelial CSCs in vitro and inhibits their metastatic colonization abilities in vivo; however, transiently inducing EMT at primary site enhances local invasion and blood entry [et al., showed that only a mixture of EMT and non-EMT HCPC-1 cells can form lung metastases in nude mice and that neither EMT nor non-EMT HCPC-1 cells establish lung metastases when injected separately [CAFs are a major type of stromal cells in the tumor microenvironment . There aude mice . Anotherude mice . Althougod entry , 68. Tsuparately . In addiAccumulating evidence suggests that MSCs or MSC-derived cells play important roles in tumor growth and metastasis. MSCs are present in various organs in human adults; however, the major cell origin of MSCs in the tumor microenvironment is bone marrow. Bone marrow-derived MSCs have different cell fates in the tumor microenvironment in which they can differentiate or transdifferentiate into CAFs, endothelial-like cells, pericytes and macrophage-like cells . Bone maversus heterotopic xenograft mouse models and in vitro differentiation analyses [in vitro as well as contribute to wound healing in vivo [in vitro [in vitro [Because CSCs have multi-lineage differentiation abilities, CSCs might be one of the sources of MSCs and MSC-derived cells. GSCs have mesenchymal-lineage differentiation abilities in orthotopic analyses . Subcuta in vivo . Our datin vitro . Small lin vitro . PC-3 anin vitro . Individin vitro . These oin vitro . Therefoin situ as well as facilitate tumor invasion and dissemination [in vitro and in vivo; however, whether CSCs in non-brain tumors can transdifferentiate into neurons or other neural lineages remains unclear. Some cancer cells display components of the phenotypes of neurons; for example, breast-to-brain metastatic cells exhibit GABAergic characteristics that are similar to neuronal cells to promote their metastatic growth [in vitro. WER-RB1 CSCs differentiate into neuron-like cells in neuronal-medium and express GFAP in vitro [in vitro induces a neuron-like phenotype as well as the expression of multiple neural cell markers BM88 and NF-200 [in vitro [in vitro, there is still insufficient direct evidence to demonstrate that CSCs generate functional neuron-like cells in the tumor microenvironment in vivo.Nerves are a common component of the tissue microenvironment and they play important roles in regulating cell behaviors and tissue homeostasis; however, their roles in tumorigenesis and metastasis are poorly understood. Different groups have reported that the nervous system contributes to tumor growth and metastasis . Autonommination . Embryonc growth , indicatc growth . Severalin vitro . In addid NF-200 . Ovarianin vitro . Because+ hematopoietic cells has also been observed in ovarian carcinoma. These cell fusion-derived epithelial cancer cells co-express epithelial and hematopoietic cell markers and exhibit elevated stemness and migratory abilities [Helicobacter infection, which induces chronic inflammation, BMDCs home to the stomach and initiate tumorigenesis through metaplasia and dysplasia to intraepithelial cancer in a cell fusion-dependent manner [in vitro and in vivo.The tumor microenvironment contains innate immune cells  and dendritic cells) and adaptive immune cells (T and B cells) , 86. Thebilities . Cell fut manner . In thist manner , 97. MDSt manner . As a rein vitro [in vitro [in vitro and generate multiple germ layer tissues in vivo. Moreover, endodermal ductal structure generated by this iPS cell line in vivo develops into pancreatic intraepithelial neoplasia in 3 months and progresses into invasive carcinoma by 9 months, indicating that cancer iPS cells mimic tumor initiation and progression in vivo [Cancer can be regarded as a disease of reprogramming and differentiation , 100. Diin vitro . Miyoshiin vitro . GBM neuin vitro  and sarcin vitro  have alsin vitro . This pa in vivo . In addi in vivo . TherefoTumors can be considered as a complex organ that contains heterogeneous cancer cells and other types of stromal cells; these cells, together with the ECM and local and systemic factors, construct the tumor microenvironment. It is well-defined that cancer cells recruit multiple types of stromal cells from local tissues or bone marrow to construct their own microenvironment . The genin vitro and in vivo, which strictly follows the CSC model. Emerging data also provide evidence that CSCs have the potential to transdifferentiate into numerous types of stromal cells in the tumor microenvironment even though most of the lineage transdifferentiation from CSCs is still unproven (Figure in vitro and in vivo. This transdifferentiation is observed in GBM, breast, ovarian and renal cancers, although the transdifferentiation efficiency is believed to be relatively low. Moreover, CSCs can generate pericytes in GBM. These findings update the basic principle of the cell origin in tumor angiogenesis. In addition, CSCs have been demonstrated to give rise to mesenchymal lineage cells, such as adipocytes, osteoblasts and osteoclasts, in vitro. Several pieces of evidence indicate that adipocytes and neurons promote tumor growth, metastasis and drug resistance in different tumors [Current evidence supports that CSCs can regenerate cancer cells n Figure . It has t tumors , 112. Osin vivo using genetic mouse models and clinical samples. Taking advantage of lineage-tracing approaches in mice will help us gain a better understanding of cell fates of CSCs. We also should determine the transdifferentiation efficiency of different cell types in different tumors. Second, understanding the molecular mechanism underlying CSC transdifferentiation is also important for deciphering tumor heterogeneity and developing new therapeutic targets. Currently, we are still confused about when and how CSCs undergo multi-lineage differentiation in the tumor microenvironment to promote tumor growth and progression. It is apparent that CSC transdifferentiation is regulated at the genetic and epigenetic levels. It is also expected that CSC transdifferentiation is tightly controlled temporo-spatially. It is urgent to determine the types of regulators and molecular switches that are responsible for this transdifferentiation scenario. These observations also lead to another thought about whether these transdifferentiation-derived cells can dedifferentiate into CSCs or transdifferentiate into other types of cells. If this is true, we will find that multi-sources of cells in the tumor microenvironment contribute to the complexity of tumor heterogeneity.Although the transdifferentiation of CSCs into tumor stromal cells is compelling and provides a new dimension explaining tumor heterogeneity, many aspects of CSC transdifferentiation remain elusive. First, CSC transdifferentiation needs to be further elucidated CSC transdifferentiation may have tremendous clinical implications in cancer treatment. Current strategies for anti-cancer therapy mainly focus on eradicating the fast-dividing cells or differentiated cells; however, these therapeutics have minimal effect on those CSCs that are slowly dividing or that remain quiescent. Increasing evidence suggests that CSCs are significantly insensitive to multiple therapeutics , 114. MoIn conclusion, the observations of the transdifferentiation of CSCs into multiple stromal cells in tumors suggest that the plasticity and functional heterogeneity in cancer cells and their stromal compartments is far more complex than we expected. Although this CSC transdifferentiation still remains elusive in many aspects, the discoveries in this field will advance our understanding of the plasticity and heterogeneity of tumors and may help with developing new diagnostic and therapeutic strategies against tumors.


Tobacco use is the leading cause of preventable disease and death in the United States . IncreasThe Tobacconomics ProgramAs of January 1, 2014, a total of 48 states (including DC) applied cigarette tax stamps. Only four of these authorized the use of high-tech stamps. Three of these four states  have implemented their use; New Jersey has not . Of the Although Native American tribes within the United States are protected by sovereign immunity and states do not have legal authority over tribes within their borders, agreements, such as ones to regulate tobacco sales, may be negotiated. Thirty-four states have federal reservation land within their borders. Of these, 20 regulated tribal tobacco sales as of January 1, 2014, 13 of which explicitly addressed stamping of products sold on-reservation . Of thosThis report indicates that although the majority of states required low-tech cigarette tax stamps as of January 1, 2014, few were using high-tech stamps, applying stamps to other tobacco products, or working with tribes on stamping agreements. Depending on analytical approaches and definitions of illicit trade, it is estimated that 8%–21% of cigarettes consumed in the United States are purchased illicitly  did not require any stamps, making tax collection more difficult and potentially facilitating illicit trade. The majority of states use low-tech stamps on cigarettes, which are easier to counterfeit . These cA few states are successfully employing high-tech stamps . Anti-coAlthough most states applied at least low-tech stamps to cigarettes, only a few expressly stamped little cigars or RYOT. Requiring stamps on other tobacco products, especially cigarette analogues such as little cigars and RYOT, is an important aspect of preventing tax avoidance by minimizing opportunities and incentives for substitution . WithoutA critical facet of a comprehensive approach to tobacco stamping is the inclusion of all sources of tobacco in this practice, including sales by Native American tribes. Several states have entered into agreements with Native American tribes on general tobacco-related issues or have negotiated specific tax agreements with tribes to reduce the avoidance of tobacco excise taxes by nonmembers, including application of tax stamps to products sold on-reservation. Although tribal members who purchase tobacco on-reservation are exempt from state taxation, nonmembers purchasing on-reservation are not exempt from state taxation; these illegal purchases by nonmembers are a significant source of illicit trade because of challenges in collecting taxes on sales to nonmembers . AgreemeWhat is already known on this topic?Increasing the unit price on tobacco products is the most effective tobacco prevention and control intervention, especially among price-sensitive populations, such as youth. Illicit tobacco trade can undermine the effectiveness of high tobacco prices by providing tobacco users with cheaper priced alternatives. Tobacco tax stamping is intended to further support efforts to prevent and reduce illicit trade.What is added by this report?A comprehensive tax stamping approach includes the use of digital, encrypted (“high-tech”) stamps, the application of stamps to all tobacco products, including little cigars and roll-your-own tobacco; and working with Native American tribes on stamping agreements. As of January 1, 2014, most states used traditional paper (“low-tech”) stamps that are easy to counterfeit, and many did not explicitly require stamps on cigarette-equivalent products such as little cigars and roll-your-own tobacco. Approximately two thirds of states with federal reservation land did not have codified agreements that permit tobacco stamping of tribally sold products.What are the implications for public health practice?Illicit trade undermines tobacco control efforts and might contribute to health disparities. Comprehensive tax stamping policies could enhance U.S. efforts to reduce illicit trade, thereby increasing revenues as well as protecting public health and reducing smoking by stopping illegal cigarette sales.The findings in this report are subject to at least three limitations. First, the cigarette, little cigar, and RYOT data were limited to codified statutory and administrative law and do not include Attorneys General opinions, case law, or departments of revenue–issued notices, rulings, or decisions. For example, California’s statutes or regulations do not explicitly call for little cigar stamping. However, per a notice issued by California’s Board of Equalization , all little cigars must be stamped.A comprehensive approach to tobacco tax stamping could be an important tool in reducing illicit trade and revenue loss in the United States. Applying tax stamps to all tobacco products, and for those states with federal reservation land within their borders, working with tribes to negotiate mutually beneficial agreements, including the use of stamps on tobacco products sold on reservation land, could have an important impact on reducing illicit trade and further reduce smoking and associated health care costs as well as recoup lost revenues from illicit trade . Additio


The Translational Bioinformatics Conference (TBC) has been one of the most successful multidisciplinary conference series in the rapidly emerging field of translational bioinformatics. The Third Annual TBC (TBC 2013) jointly held with the Human Genome Variation meeting (HGV 2013) for 5 days at the JW Marriot Hotel, Seoul, South Korea provided the opportunity to improve the understanding of complex and rare diseases and propose new ways of approaching basic health problems by integrating genomic and clinical data.There is growing demand for translational bioinformatics approaches that would allow the heterogeneous data from basic research to be integrated systematically into clinical practice in a cohesive and large-scale manner. The expectation is for translational bioinformatics to integrate and correctly extract clinically actionable information from huge data sets at all levels across biological molecules, subcellular compartments, signaling pathways, cells, tissues, organs, and systems. Another challenge is the cultural differences between research scientists and clinicians and their different value systems.Linking molecular research and clinical applications will enormously benefit human health. Applying translational bioinformatics to next-generation cancer genome sequencing has been very successful in extracting valuable prognostic gene signatures, clinically actionable mutations, novel network biomarkers, and therapeutic targets across many types of malignancy, with examples appearing in areas such as pharmacogenomics, drug developments, and rare Mendelian disorders. I believe that novel translational bioinformatics methodologies developed by multidisciplinary stakeholders will rapidly expand into many clinically disease areas.Personalized medicine involves determining individualized therapies based on the genomic and clinical profiles of specific individuals, which includes utilizing their molecular data. While the predictive power of a plethora of genomic signatures has been successfully demonstrated, dichotomizing phenotypes based on statistical significances is problematic due to the presence of major heterogeneity. In an attempt to realize truly personalized diagnostics and prognostics, Gardeux et al.  have proposed a method of mechanism-level analysis applied to single pairs of samples, such as tumor vs. matched control, primary tumor vs. metastases, and before vs. after treatment. Their N-of-1-pathways predictions not only outperform conventional methods such as Gene-set Enrichment Analysis (GSEA) but also identify unique sample/patient mechanisms, which is a requirement for personalized medicine [Patient-derived xenografts in mouse models have been widely used in the testing of new anticancer drugs in preclinical evaluations of experimental therapeutics due to them representing an approximation to the clinical characteristics of patients. These models involve transplanting human cancer cells into mouse host tissues. However, as Yang et al.  pointed out , profiliBiomarkers are indicators of biological states that are useful for the evaluation of physiological states, pathogenic processes, and therapeutic responses. While traditional biomarkers examine the current disease status mainly based on a single molecule, Li et al.  focused on the development of phase-specific biomarkers for the progression of prostate cancer through an integrative analysis of gene expression profiling and protein interaction networks . They prShin and Nam  have expanded the concept of biomarkers into translational biomarkers . Their pShin and Nam proposed a model that predicts the survivability of breast cancer patients using a predictor module, and then calculates the variable importance of the prognosis factors using a descriptor module, which also makes it possible to separate patients with similar prognostic profiles. They successfully demonstrated the translational power of the proposed method on the SEER  cancer incidence database , which iFor discriminating disease phenotypes and discovering meaningful biomarkers, Han  proposed applying derivative component analysis (DCA) to high-dimension serum proteomics data . CompareThe thousands of genetic variants discovered by genome-wide association studies (GWASs) have been shown to explain only a very small proportion of the underlying genetic variance of complex traits. Gene-gene interaction analysis is expected to unveil a significant proportion of missing heritability information about complex traits. Kwon et al.  have developed an entropy-based algorithm, IGENT (Information theory-based GEnome-wide gene-gene iNTeraction) , that suDespite the recent advances in GWASs and next-generation sequencing technologies associated with the discovery of thousands of mutations and polymorphisms, the causal relationship between pathophysiological molecular mechanisms and therapeutic responses of complex diseases remains unclear. Public databases and numerous biomedical knowledge resources are invaluable when performing translational bioinformatics research. Prosperi et al.  have attempted to fill this phenome-genome gap by applying a spectrum of linear and nonlinear machine-learning methods to a large clinical and genomic attribute set involving complex clinical conditions: asthma, wheeze, and eczema . Grover Improved bioinformatics tools and methods are required for successful translational research. Advanced solutions have been introduced for well-known bioinformatics problems. Wang et al.  have developed a graphics-processing-unit-accelerated massively parallel computing algorithm for miRNA target identification that was 166 times faster than previous methods . Joung eThe increasing number of multi-institutional collaborations and demand for data integration makes the detection and resolution of measurement-unit conflicts challenging. Samadian et al.  utilized existing ontologies and standards for scientific data representation to build a Semantic Web Service-based approach to automatic measurement-unit harmonization . The traThe meeting, TBC 2013, has provided the opportunity for translational bioinformatics researchers to come together and substantially improve the understanding of biomolecular and pathophysiological mechanisms, which will contribute greatly to the development of truly personalized diagnostics, prognostics, and therapeutics. I congratulate the speakers and authors at this conference who are shaping the future of how biomedical informatics translates into better clinical practice. Many health topics are increasingly within the scope of translational bioinformatics, including rare and complex human diseases, cancer, biomarkers, pharmacogenomics, drug repositioning, and clinical decision support systems. As Alan Kay said, the best way to predict the future of translational bioinformatics is to invent it.The authors declare that they have no competing interests.


We tested the hypothesis that early-life developmental exposures  affect the balance of investment between acquired and innate immune defenses in adults. Analyses of two immuno-markers suggest greater nutrition and pathogen exposure, and lower extrinsic mortality cues associated with a small bias towards acquired defenses. Life history theory predicts resource allocation trade-offs between competing functions and processes. We test the hypothesis that relative investment towards innate versus acquired immunity in humans is subject to such trade-offs and that three types of early developmental exposures are particularly salient in shaping adult immunophenotype: (i) pathogen exposure, (ii) nutritional resources; and (iii) extrinsic mortality cues.We quantified one aspect each of innate and acquired immune function, via C-reactive protein and Epstein–Barr virus antibodies, respectively, in a sample of 1248 men and women from the Philippines (ca. 21.5 years old). Early developmental exposures were assessed via long-term data collected prospectively since participants’ birth (1983–4). We calculated a standardized ratio to assess relative bias towards acquired versus innate immune function and examined its relationship to a suite of predictors via multiple regression.In partial support of our predictions, some of the measures of higher pathogen exposure, greater availability of nutritional resources, and lower extrinsic mortality cues in early life were associated with a bias toward acquired immunity in both men and women. The immune profile of women, in particular, appeared to be more sensitive to early life pathogen exposures than those of men. Finally, contrary to prediction, women exhibited a greater relative investment toward innate, not acquired, immunity.Early environments can exert considerable influence on the development of immunity. They affect trade-offs between innate and acquired immunity, which show adaptive plasticity and may differ in their influence in men and women. Individual differences in immune function have received considerable public health and clinical attention owing to the rising prevalence of chronic inflammation and atopic diseases in affluent industrialized populations , but a mThese threads converge within the emerging discipline of ecological immunology (ecoimmunology), which places particular emphasis on how ecological factors affect an individual’s immunophenotype and thus places an organism’s investment in immune defenses within the context of local environmental conditions , 13. SucPrevious work in human ecoimmunology has focused primarily on trade-offs between overall investment toward immune function and other energetically costly processes such as growth and reproduction , 21–25. The costs and benefits of subsystems of immune defense are variable, with the optimal allocation of resources across different arms of immunity depending on the specifics of the local ecology , 27–29. First, we predict that individuals growing up in a high-pathogen environment will prioritize investing in specific, efficient acquired immune defenses, all else being equal. The long-term benefits of investing in development of acquired immunity will increase with the frequency of encountering pathogens repeatedly in one’s environment. Conversely, if early life pathogen exposure is low, this would reduce the potential long-term payoffs of investing in costly development of acquired immunity and favor an immune profile biased toward non-specific defenses .Second, we predict that resource availability will also affect the optimal investment to the two arms of the immune system. Although specific, acquired immune defenses provide clear advantages in a high-pathogen environment, the organism’s ability to invest in specific immune development can be constrained if nutritional resources during early development are scarce. Thus, we predict that greater resource abundance in the prenatal and early post-natal period will allow increased investment in acquired immunity, given its relatively high developmental costs. Conversely, if early-life nutritional resources are limited, this will constrain the organism’s ability to invest in the development of acquired immunity. Later in life, such an individual will exhibit an immunophenotype that relies to a greater extent on innate, rather than acquired, defenses, even if nutritional conditions in adulthood are more favorable than during the developmentally critical early-life period when we predict that biases in immune allocation are set .Finally, we predict that as the potential risk of unavoidable (extrinsic) mortality increases, the optimal pattern of immune investment will shift to favor non-specific immunity. Under such conditions, the high upfront costs of acquired immunity would receive a lower priority, as shorter lifespan reduces the long-term benefits of acquired defenses. Conditions that may signal higher risk of extrinsic mortality  would therefore be associated with a shift in resource allocation toward immune function prioritizing the less costly in the short-term option, i.e. innate immunity. Cues of extrinsic mortality have been shown to shift developmental priorities of investment in other domains, e.g. from investment in somatic growth to early reproduction , and we We test these predictions using data from a prospective longitudinal study of humans in the Philippines. Using two markers of immune function, C-reactive protein (CRP) and Epstein–Barr virus (EBV) antibodies, we assess relative investment toward innate and acquired immune defenses, respectively, in a large cohort of ca. 21.5-year olds. With data on their early-life environments , we examine whether their adult immunophenotype exhibits patterns consistent with the hypothesis of early developmental plasticity and trade-offs across branches of immune function . To placN = 1453. Because one of our outcomes (CRP) is elevated acutely during infections, participants reporting infectious symptoms during the interview were also excluded (N = 205). Median CRP was 0.7 mg/L for the participants reporting infectious symptoms, compared with 0.2 mg/l for the rest of the sample. Although menstrual phase is also known to affect CRP levels in women [N = 414), we did not find significant differences in CRP levels between women in the early follicular, late follicular, or luteal phase of the cycle . A total of 1248 healthy, non-pregnant individuals (men and women) contributed data to these analyses. All data were collected under conditions of written informed consent using protocols approved by the Institutional Review Board of the University of North Carolina, Chapel Hill. Data from the CLHNS are publically available from the Dataverse Network  of the Odum Institute at the University of North Carolina, Chapel Hill: http://arc.irss.unc.edu/dvn/dv/cebu.The Cebu Longitudinal Health and Nutrition Survey (CLHNS) began in 1983–4 with the recruitment of 3327 pregnant women from randomly selected urban and rural neighborhoods in Metropolitan Cebu City, Philippines . The stuin women , prelimiTo assess the balance of investment toward innate versus acquired immunity, we measured two biomarkers of immune function: CRP (innate) and EBV antibodies (acquired immunity). CRP is an acute phase protein and biomarker of inflammation and an important component of innate immunity. Produced primarily by hepatocytes in response to cytokine signals, CRP activates complement, promotes activity of phagocytic cells, and opsonizes bacteria, fungi, and parasites . ConcentThe EBV is a ubiquitous herpesvirus that infects nearly 90% of adults in industrialized nations, while infection rates approach 100% during the first 5 years of life in lower income nations , 42. Oncr = 0.03, N = 1248, P = 0.28). To evaluate the relative investment toward innate versus acquired immunity, we constructed a ratio between the two variables. Because 36.1% of samples (N = 450) had CRP levels below assay sensitivity, we re-coded all zero values as 0.1. We then log-10 transformed and standardized  the values of CRP and EBV to allow a direct comparison. Because higher levels of EBV antibodies indicate reduced cell-mediated immune function, we multiplied log-EBV antibody titers by −1 (i.e. reverse-scoring) so that higher values would indicate higher levels of cell-mediated immunity. We then calculated the ratio of standardized acquired (cell-mediated) immunity  to standardized innate immunity  as the CMI:CRP ratio. A value of 1.0 represents equal investments in both aspects of immunity, relative to the sample distributions. Higher values represent bias toward acquired immunity, while lower values represent a bias toward innate immunity.Across all individuals, CRP levels were not significantly correlated with EBV antibody titers (N = 341). These CRP values were positively and significantly correlated . Even though the correlation was not particularly strong, these data nevertheless suggest that over time inter-individual differences in CRP remained somewhat consistent. At the age 14–15 years, a subset of participants received a typhoid vaccine, and 47.9% of the sample responded to vaccination with a ≥4-fold increase in anti-typhoid antibody titer [N = 36), indicating reduced cell-mediated immunity. Vaccine responders had lower levels of EBV antibodies , indicating higher cell-mediated immunity. The difference in EBV antibodies across the groups approaches significance  and is consistent with the interpretation that individual differences in overall investments in acquired immune defenses are relatively stable in this sample.To evaluate the intra-individual consistency/tracking (within the population) of our measures of innate and acquired immunity, we compared immune responses in a sub-sample of study participants, measured at two time points. Concentrations of CRP were measured during adolescence (14–15 years of age) and young adulthood (21–22 years of age) in a subsample of study participants  and including them reduced overall model fit, we did not consider them further in analyses.Initially, we also explored the role of three measures of socioeconomic status (SES), assessed both at birth (for the mother) and at the age of sample collection in young adulthood , as described previously . BecauseWe conducted model validation by checking for multicollinearity among predictor variables  and by b = 0.26, SE = 0.08, P = 0.002), but early developmental exposures were not associated with total immune investment .We first considered whether infectious, nutritional, and extrinsic mortality risk variables predicted the total level of immune investment (sum of the CMI and CRP standardized scores). Males had significantly higher values than females , but no such relationship was present among males . Females also had significantly lower CMI:CRP ratio than males  to 1.36 . After excluding all non-significant interactions, we obtained a more parsimonious model  but not males  had higher CMI:CRP ratio if they were born during the dry season compared to the wet  was involved in a significant interaction with sex , only birthweight was significantly associated with adult immune phenotype. As predicted, individuals with higher birthweight had higher adult CMI:CRP ratio, suggesting greater investment toward acquired immune defenses. Of the three measures of early-life pathogen exposure , two contributed to explaining the variance in adult CMI:CRP ratios, both in line with our predictions. First, individuals who experienced greater likelihood of exposure to animal feces in the first year of life showed higher CMI:CRP ratios. Second, among women but not among men, individuals born during the dry season  exhibited higher CMI:CRP ratios. Finally, of the three measures of early-life psycho-social stress , which can be seen as cues of extrinsic mortality, only one was significantly associated with adult immunophenotype. Individuals who experienced maternal absence in childhood showed lower adult CMI:CRP ratio, as predicted. In sum, these results provide partial support for the existence of trade-offs between the two arms of the immune system and indicate these trade-offs are affected by some early-life developmental factors, as predicted in our recent synthesis on this topic . AlthougThe majority of ecoimmunology research on within-system trade-offs in immune development has thus far centered on relatively small-bodied and, by comparison to humans, shorter-lived species. Trade-offs between the arms of the immune system in such organisms are more likely to be pronounced, and indeed detected, as smaller animals do not have sufficient energy reserves to rely on as buffers/capital, in the way that larger-bodied animals do . NeverthThe importance of early-life pathogen exposure in potentially affecting levels of inflammation in young adulthood in this sample is already known: higher levels of pathogen exposure in infancy predicted reduced CRP in young Filipino adults , a findiExisting data on inter-individual variation in measures of acquired immunity are also consistent with the logic of our framework . ChildhoN = 74), individuals that did not respond with antibodies to typhoid vaccination showed much higher CRP levels in adulthood [N = 1248). Our framework [Previous research in human ecological immunology has not explicitly tested the existence of trade-offs between the two branches of the immune system but some results are suggestive. Data from Cebu have shown the possible existence of such trade-offs between acquired and innate defenses: in a small subsample of subjects (dulthood . Our curramework  thus extP = 0.06) and overall morbidity in the first 2 years of life  than women is similarly unexpected as extensive existing data point toward women having a consistent immunological advantage over men . WhetherOur analyses have focused on trade-offs between investments in innate and acquired immunity, but it is important to note that both systems are necessary for survival, and that in practice they work together to provide a relatively seamless network of defense. As such, one might expect small differences in relative levels of investment in relation to developmental environments. Indeed, the differences we find here are small and predictors in statistical models explain a tiny fraction of the variance CMI:CRP ratios . This coIt is also notable that when we examined the effects of early life exposures on total immune investment (CMI + CRP), none of the variables we considered had any explanatory power. This may be because early life exposures in this cohort were relatively benign and they did not reach a threshold beyond which they would have had an adverse effect on overall investment toward immune defenses. The weak trade-offs within the immune system we documented, however, suggest that some adjustment in allocation to resources within the immune system may occur even when overall ability to invest toward immune defenses is robust. This interpretation is consistent with the observed positive association between CMI:CRP ratio and total immune investment (CMI + CRP): individuals with greater overall investment in immune defenses prioritize acquired immunity over innate. Even small variation in resource availability may therefore be associated with shifts in the optimal trade-off level between acquired and innate defenses.in utero, the wide range of available variables, and the large sample size, are significant strengths of this study. Our findings should nevertheless be regarded as preliminary, and generative of future tests and refinements of the proposed framework for within-immune-system trade-offs [It is important to note that neither CRP nor EBV, measured on their own are sufficiently representative of innate and acquired immune function to provide us with definitive conclusions regarding trade-offs within the immune system. Both arms of the immune system comprised multiple pathways that interact in complex fashion. In addition to potentially being affected by female reproductive function, CRP levels may have also been related to subclinical inflammation processes that would not have been recognized from self-reported measures of morbidity. EBV, on the other hand, has been shown to be affected by current psycho-social stress , anotherade-offs  based on


Up to now, immunization of disease propagation has attracted great attention in both theoretical and experimental researches. However, vast majority of existing achievements are limited to the simple assumption of single layer networked population, which seems obviously inconsistent with recent development of complex network theory: each node could possess multiple roles in different topology connections. Inspired by this fact, we here propose the immunization strategies on multiplex networks, including multiplex node-based random (targeted) immunization and layer node-based random (targeted) immunization. With the theory of generating function, theoretical analysis is developed to calculate the immunization threshold, which is regarded as the most critical index for the effectiveness of addressed immunization strategies. Interestingly, both types of random immunization strategies show more efficiency in controlling disease spreading on multiplex Erdös-Rényi (ER) random networks; while targeted immunization strategies provide better protection on multiplex scale-free (SF) networks. The structure and dynamics of multiplex networks have attracted much attention by the scientific communities et al. unfolds the prevalence of the disease in both layers, even if the epidemiological parameters are assigned values lower than the epidemic threshold of each layer. By superposition processing of the network layers, Zhao et al. show that a strong positive degree-degree correlation of nodes in different layers could lead to a clearly low epidemic threshold and a relatively smaller infection size Recently, an increasing number of works has tried to understand the dynamics of epidemic spreading in multiplex networks. Along this line, various mechanisms aiming at exploring the disease propagation process in multiplex topology are proposed and investigated. Examples include competing epidemics As above described, though there have been some achievements focusing on the effect of multiplex architecture on the epidemic dynamics and the resulting threshold, the impact of such increased complexity on the immunization strategies is still virgin Here, with the SIR epidemic model on multiplex networks For simplicity , we consider the SIR dynamics as the epidemiology model, and then inspect the effect of immunization strategies on disease propagation in multiplex networks. With regard to networks, we select multiplex Erdös-Rényi (ER) random networks For networked immunization, there is usually one critical index, immunization threshold Multiplex node-based immunization refers to the case that a fraction of multiplex nodes is random or targeted immunized. If we use where where Then, the probability where Thus, the existence of epidemic regime under multiplex node-based immunization requires the largest eigenvalue whereandFor multiplex node-based random immunization, each node has the same probability to be immunized, so we can write For multiplex node-based targeted immunization, the nodes are generally immunized according to their degree, betweeness or Under this immunization framework, the immunized probability of a multiplex node with spreading degree where where orks see .Besides above proposal, the objects of immunization now turn to layer nodes. If we define The generating function of remaining joint degree distribution by following a randomly chosen link of network layer For layer node-based immunization, the probability Similarly, the existence of epidemic regime requires the largest eigenvalue whereandFor layer node-based random immunization, each node in the same network layer has equal probability of taking immunization. So, we can get where For the layer node-based targeted immunization, the immunized probability of a layer node with degree where where Thus, for layer node-based targeted immunization of duplex networks, the critical immunization threshold is given byIn d random  and targd random . TogetheSubsequently, we further extend the layer node-based immunization proposals to multiplex SF networks and compare its efficiency with case of multiplex ER networks in To sum, we propose four kinds of immunization strategies of multiplex networks, including multiplex node-based random immunization and targeted immunization, and layer node-based random immunization and targeted immunization. By using generating function methods, we provide one new theoretical framework which allows us to accurately calculate the critical immunization thresholds of different immunization strategies. We also evaluate the efficiency of the proposed immunization strategies for multiplex ER networks and multiplex SF networks. We show that both multiplex node-based and layer node-based random immunization has higher efficiency in multiplex ER networks, while two types of targeted immunization strategies can provide better protection in multiplex SF networks.


Our general aim is to better understand the biophysical and biochemical principles that govern nascent or stress-destabilized proteins to unfold and re/fold to their native state or, alternatively, choose to misfold and aggregate into cytotoxic species, which in animals can cause degenerative diseases.in vitro protein refolding assays are typically low and a large fraction of the polypeptide precipitates into insoluble aggregates.When a protein is first artificially unfolded by a chaotropic agent, it may thereafter spontaneously refold into its three dimensional native state without necessitating additional steric information from other macromolecules in the solution. Anfinsen , thus maLiving cells cannot accumulate high concentrations of chaotropic agents to specifically unfold potentially toxic aggregates, while leaving intact the surrounding native proteins. Instead they express molecular chaperones that can specifically recognize and bind with high-affinity to the misfolded polypeptides, which are to be unfolded into intermediates that can, thereafter, spontaneously refold into low-affinity native products (Hinault et al., In the last two decades, a major discovery has been that misfolded and aggregated conformers, even from “normal” non-toxic proteins, can become cytotoxic and infectious (Ben-Zvi and Goloubinoff, It is not clear why some early forms of aggregated proteins are toxic to mammalian cells, especially to neurons, where they apparently induce apoptosis. It is suggested that protofibrils may spontaneously form holes in membranes and disturb biological membranes in general (Lashuel and Lansbury, Even without aging and abiotic stresses, when polypeptides are synthetized in the ribosome, growing chains encounter the highly crowded environment of the cytoplasm, which is composed of up to 200 mg/ml proteins (Finka and Goloubinoff, de novo synthesized polypeptides may readily fold to the native state, as initially shown in vitro by Anfinsen in the simple case a monomeric enzyme. At variance, the concomitant or sequential assembly of multineric complexes may necessitate molecular chaperones to maintain near-native subunits in an assembly-competent state until other subunits are present to form functional oligomers. Polypeptides may also have to be prevented by cytoplasmic chaperones from completing folding while still in the wrong compartment and thus be maintained in a translocation-competent state poised to translocate through membrane pores to the stroma of mitochondria, chloroplasts or the lumen of the endoplasmic reticulum (Figure While emerging from the ribosome, intrinsically unfolded polypeptides may stay unfolded and, depending on the presence of osmolytes, pre-aggregated seeds or of stressful conditions, they may misfold and aggregate into toxic species as in the cases of α-synuclein or tau (Luheshi and Dobson, Depending on the severity of various abiotic stresses, such as heat-shock, or owing to destabilizing chemical modifications or mutations, polypeptides may misfold and aggregate into potentially toxic conformers (Figure Frontiers in Molecular Biosciences, Protein folding, Misfolding, and Degradation publishes original biochemical research article addressing the processes of protein synthesis, folding, misfolding, translocation, refolding, assembly, and aggregation, as well as specific mechanisms by which chemical chaperones, molecular chaperones and proteases control cellular protein homeostasis. Of particular interest are physiological cellular processes, such as de novo folding and assembly of native complexes, which do not necessarily involve chaperones preventing or reverting protein aggregation. How may chaperones differentiate bound on-pathway near-native subunits seeking to integrate native oligomers, from off-pathway oligomer-orphan subunits that become rapidly degraded?in vivo imaging of protein misfolding and amyloid accumulation in degenerative tissues and whole organisms.We are primarily interested in biochemical and molecular mechanistic studies addressing how chaperones and proteases use ATP hydrolysis to unfold, disaggregate and promote the native folding or the degradation of stably misfolded and aggregated proteins species and about the principles governing partitioning between the “rehabilitation” of misfolded proteins by unfolding and disaggregating chaperones, and “recycling” of misfolded proteins by unfolding proteases (Suraweera et al.,


Recently, applications of optical coherence tomography angiography (OCTA) have been limited to the retina and posterior segment. Although early studies have described its use for other clinical applications, its role in anterior segment vasculature and optic disc imaging has been limited thus far.We describe a novel clinical application of OCTA in a patient with dural carotid-cavernous sinus fistula (CCF), which was complicated by increased intra-ocular pressure (IOP). In this case report, we used the OCTA to delineate increased epsicleral venous flow in the affected eye with secondary raised IOP. Current measurements of episcleral venous pressure are either invasive or provide highly variable results, thus the OCTA may have the potential to provide a more reliable approach to assess episcleral vasculature. We also describe the use of OCTA to detect early glaucomatous nerve damage, associated with focal reductions in peripapillary retinal perfusion.We present an early report of using OCTA of the anterior segment to allow rapid, non-invasive delineation of abnormal episcleral venous plexus secondary to dural CCF. The OCTA was also useful for detecting early reduction in peripapillary retinal perfusion, which suggests early glaucomatous optic neuropathy. This suggests that OCTA may have a role for determining risk of glaucoma in patients with CCF in future. Anterior segment angiography has a variety of clinical applications, which may range from evaluation of corneal vascularization , to scleSince current OCTA systems are optimized for the retina and optic disc, we had previously described a technique adapted to perform scans in the anterior segment for normal corneal and limbal vessels . CarotidWe present a case of a 54-year-old Chinese man with a history of treated hypertension and diabetes mellitus, referred for suspected spontaneous left dural CCF. The patient complained of a chronically red left eye and double vision in the left gaze. Initial examination disclosed best corrected visual acuity 20/20 in the right eye and 20/25 in the left eye, associated with a left relative afferent pupillary defect and severely reduced color vision on Ishihara testing in his left eye only. Humphrey visual field assessment disclosed diffuse defects in the left eye, but was normal in the right eye. Slit lamp examination disclosed a red left eye with dilated corkscrew vessels, but no evidence of angle closure, blood in the Schlemm canal, or other causes of raised intraocular pressure. Intraocular pressure (IOP) was 28 mmHg in the left eye, and 14 mmHg in the right, healthy eye. Fundoscopy disclosed dilated retinal veins on the left side, but no venous stasis retinopathy, or choroidal detachment. The remainder of the ophthalmic examination disclosed mild left abduction deficit, mild ptosis and 3 mm proptosis on the left side. Partial embolization of the angiographically confirmed left dural CCF resulted in incomplete clinical recovery: despite complete regression of the left optic neuropathy and of the left abduction deficit, the left eye remained red, associated with raised intraocular pressure requiring topical medication. Optical coherence tomography angiography of the anterior segment was performed, using a described technique , with opThe episcleral venous outflow is a key component of intraocular pressure, as embodied by the modified Goldmann equation, which can be thought of as the pressure required to move aqueous humor out of the eye through the resistance of the trabecular meshwork and the pressure in the episcleral veins . OpticalOne of the main presumed mechanisms for raised IOP associated with CCF is increased episcleral venous pressure . ObjectiIn summary, this early clinical report describes, for the first time, the OCTA features in dural CCF, suggesting that OCTA allows rapid, non-invasive, in-depth visualization of abnormal episcleral vasculature in CCF. Further quantitative studies are needed to assess the link between abnormal episcleral vasculature and the risk for glaucoma in cases of dural CCF.OCTA, optical coherence tomography angiography; OCT, optical coherence tomography; CCF, carotid-cavernous sinus fistula; IOP, intraocular pressure


Posterior reversible encephalopathy syndrome (PRES) is an unusual condition typified by acute visual impairment caused by sudden, marked parieto-occipital vasogenic edema. Thought to be inflammatory in origin, it has been described in patients undergoing chemotherapy, with autoimmune disease, and in some infections. We report a case of PRES that occurred one week after an episode of acute pancreatitis in an otherwise healthy 40-year-old female. There was progressive visual impairment over a 24-hour period with almost complete visual loss, with characteristic findings on magnetic resonance imaging. After treatment with steroids, the visual loss recovered. Clinicians should retain an index of suspicion of this rare condition in patients with visual impairment after acute pancreatitis. A 40-year-old female presented to our emergency department with sudden visual loss over a 24-hour period. She was otherwise healthy, but had been admitted two weeks previously with an episode of acute pancreatitis secondary to alcohol intake from which she had recovered uneventfully, and without any obvious sequelae. An urgent magnetic resonance imaging (MRI) scan was performed. This revealed symmetrical areas of hypoattenuation in both posterior parieto-occipital and cerebellar regions , 2. She PRES is extremely rare, and usually diagnosed by a history of sudden visual impairment in the presence of specific radiological changes on MRI. Bilateral symmetrical hypodensitities in the parieto-occipital areas and cerebellar hemispheres on imaging are characteristic. The condition has been associated with chemotherapy, hypertension, infection and autoimmune disease.1It is thought to occur from temporary impairment of the blood brain barrier causing vasogenic edema with symptoms of reduced consciousness, seizures, headaches, and typically visual problems.Our case is unusual, as PRES caused by pancreatitis has only been reported in very sick patients with other comorbidities. It probably occurred in this case as a result of the systemic inflammatory response.6Whilst pancreatitis itself can be life threatening, this case reminds clinicians of unusual complications that can occur after discharge of patients who seem to have recovered from the disease.


BRCA mutations carriers, with an increased risk for developing high-grade serous ovarian cancer, elect to undergo risk-reducing surgery. While prophylactic surgery is associated with a significant reduction in the risk of cancer development, it also results in surgical menopause and significant adverse side effects. The development of efficient early-stage screening protocols and imaging technologies is critical to improving the outcome and quality of life for current patients and women at increased risk. In addition, more accurate animal models are necessary in order to provide relevant in vivo testing systems and advance our understanding of the disease origin and progression. Moreover, both genetically engineered and tumor xenograft animal models enable the preclinical testing of novel imaging techniques and molecularly targeted therapies as they become available. Recent advances in xenograft technologies have made possible the creation of avatar mice, personalized tumorgrafts, which can be used as therapy testing surrogates for individual patients prior to or during treatment. High-grade serous ovarian cancer may be an ideal candidate for use with avatar models based on key characteristics of the tumorgraft platform. This review explores multiple strategies, including novel imaging and screening technologies in both patients and animal models, aimed at detecting cancer in the early-stages and improving the disease prognosis.The majority of high-grade serous ovarian carcinoma cases are detected in advanced stages when treatment options are limited. Surgery is less effective at eradicating the disease when it is widespread, resulting in high rates of disease relapse and chemoresistance. Current screening techniques are ineffective for early tumor detection and consequently,  Ovarian cancer is the most lethal gynecological malignancy, with the majority of high-grade serous carcinoma (HGSC) cases being diagnosed in late stages of disease [for a comprehensive review see Bast ]. The cuBRCA family of tumor suppressor genes, which confer an increased risk for developing breast and ovarian cancers  can provide a highly detailed three-dimensional analysis of the tumor micromorphology, vascularization, and accurately quantify relative blood volumes (rBV) in tumors, which can further inform treatment plans . Traditional TVS technologies are being replaced by contrast enhanced transvaginal sonography (CE-TVS) and transvaginal color Doppler sonography (TV-CDS), which can better depict the tumor morphology by analyzing its microvasculature . In a stnt plans . Most imnt plans .ex vivo. For the purpose of reducing toxicity and to facilitate clinical translation, further research will need to focus on nanoparticles that biodegrade within the mammalian circulatory system  and HER-2 as targeted agents for tumor-specific fluorescence imaging. Thus, FR-α is overexpressed in the majority of epithelial ovarian cancers, especially in HGSC tumors with a high risk of recurrence. Increased FR-α expression is detected not only in primary tumors but also in metastatic foci and recurrent tumors . Most imnografts . PPF injnografts . The uniin vivo in both pancreatic and ovarian cancer models as an alternative to ultrasound, CT, and MRI scans  has been generated, which recapitulates key genetic alterations  and precursor lesions (STICs) that are hallmarks of the human disease  have been developed, which are able to more accurately predict tumor responses to therapy. Xenograft tumor models have been used for decades to examine the behavior of various types of therapies within a living system . They caWhile GEMM and xenografts both strive to generate accurate models of human disease and their usage at times overlaps, they have individual features best suited to distinct roles in cancer research. Tumors that develop from a xenograft retain the natural genetic alterations derived from the original source , 83. ConFurther development of representative mouse models of HGSC is an urgent need for the field  and tumo+ T-cell response in both mouse models and patients and reduced the tumor ability to suppress the immune system. This bodes well for further optimization of such treatments for individual needs, particularly as an extended weekly, dose-dense carboplatin and paclitaxel regimen has been shown to be effective in heavily pre-treated, recurrent, platinum-resistant ovarian cancer patients  could prove to be more effective for both early detection and treatment. In addition, alternative strategies for tumor-specific imaging, which involve the use of FR-α-targeted fluorescent agents, HER-2-targeted magnetic iron oxide nanoparticles, or protease-specific NIRF probes, merit further investigation as selective tools for early tumor detection, monitoring, and image-guided surgery. It is worth noting that animal models are a valuable tool in ovarian cancer research for the purpose of developing and testing novel imaging technologies, biomarkers, and experimental treatments. However, there are currently a limited number of animal models available for HGSC. Advances in tumor xenograft technologies have enabled the development of personalized avatar mouse models, which have emerged as an ideal drug-testing platform, especially in concurrent clinical trials. Additionally, tumorgrafts appear to have qualities well suited to model HGSC. We suggest that avatars have the potential to improve patient outcome and quality of life by reducing the cost and toxicity of ineffective imaging and treatment.Advances in tumor screening and imaging may help determine the optimal time to employ risk-reducing surgical approaches in women at high risk for HGSC, including Anders W. Ohman, Noor Hasan, and Daniela M. Dinulescu wrote the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In vitro MSC exhibit demonstrated capabilities to differentiate into a wide variety of tissue cell types. Hence, many MSC-based therapeutic approaches have been performed to address bone, cartilage, or heart regeneration. Furthermore, prominent studies showed efficacy of ex vivo expanded MSC to countervail graft-vs.-host-disease. Therefore, additional fields of application are presently conceived, in which MSC-based therapies potentially unfold beneficial effects, such as amelioration of non-healing conditions after tendon or spinal cord injury, as well as neuropathies. Working along these lines, MSC-based scientific research has been forged ahead to prominently occupy the clinical stage. Aging is to a great deal stochastic by nature bringing forth changes in an individual fashion. Yet, is aging of stem cells or/and their corresponding niche considered a determining factor for outcome and success of clinical therapies?Multipotential mesenchymal stromal cells (MSC) are present as a rare subpopulation within any type of stroma in the body of higher animals. Prominently, MSC have been recognized to reside in perivascular locations, supposedly maintaining blood vessel integrity. During tissue damage and injury, MSC/pericytes become activated, evade from their perivascular niche and are thus assumed to support wound healing and tissue regeneration.  MSC have been isolated from embryonic tissues as well as from adult up to advanced ages   seeding densities in a defined growth medium . Besides applications in musculoskeletal defects and trauma MSC are also widely tested in pathologies, which are of immunological etiology such as graft-vs.-host disease or multiple sclerosis, lupus, diabetes type I, and Crone's disease. The reason is, MSC exhibit dominant immune-modulatory properties (Castro-Manrreza and Montesinos, In recent years, more than 500 clinical trials employing MSC for the treatment of various diseases have been registered worldwide (Many approaches are still exploratory and many ongoing clinical trials are still in Phase I Most strategies were carefully validated in diligently designed preclinical tests. However, commencing clinical trials firstly address safety issues thus rarely corroborating results cannot be expected. Early phase clinical studies also often comprise small heterogeneous groups exhibiting variant health status, age and ethnicity. Also sex-specific differences appear to be of relevance (Tajiri et al., in vitro expansion. At that point the uncertainty arises whether replicating cells accumulate damage which turns them senescent (Bonab et al., Most trials refrain from stratifying into age groups for obvious reasons of addressing a specific pathology rather than a gerontological principle. Animal trials are mostly done on young animals; clinical trials are rarely concerned with young or middle aged adults. MSC for animal experimentation are most often isolated from young animals, instead in humans autologous approaches are performed mostly in aged individuals. Further concerns are the greatly varying cell isolation and expansion procedures, as the outcome of a clinical trial is pertinently influenced by the quality of a cellular product. In fact, the population of cells harvested from bone marrow aspirates is very heterogeneous, consisting only of 0.001–0.01% mesenchymal precursor cells. To gain sufficient numbers of potential stem cells, the isolates are purified for subsequent First animal studies showed, already 10 years ago, that the transplantation of aged rat MSC was less effective (Zhang et al., It is aged people, who are the primary targets for stem cell therapy. Provided putative deviations, procedures selecting for flawless cells are potentially required before applying aged cells in autologous therapy. Allogenic transplantation of stem cells derived from young donors are considered to potentially overcome aging-related limitations Figure .It is further conceivable that the epigenetic status and/or aging as well as pathology-specific changes of cells may be different when isolating MSC from different tissues. It is currently unclear how to predict the optimal MSC source in order to warrant an optimal outcome (Golpanian et al., Microenvironmental conditions, epigenetic processes including DNA methylation, telomerase activity, microRNA as well as specific growth factor signaling actively modulate MSC fate (Madonna et al., Considering the knowledge that the patient's disease state pertinently affects MSC functionality, the impact of medication on MSC, be it before or after implantation, is largely neglected and little data are available. MSC are greatly resistant to chemotherapy agents and therefore, therapies in patients undergoing cancer treatment are conceivable (Beane et al., in vitro data regarding non-clonal culture and multi-lineage differentiation. Therefore, the true stem cell character of these MSC has yet not be fully proven (Bhartiya, in vivo are still scarcely known. Currently, information regarding biological and molecular features, as well as the dominant influences on their surrounding tissue environment, appears insufficient (Murray et al., MSC are considered “work horses” for cell-based therapy. This is because very little ethical concerns have been raised and it is now widely accepted that MSC are largely resistant to malignant transformation. Being ubiquitously present in many tissues, MSC could be successfully isolated from cord blood, fat, skeletal muscles, dental pulp, and several other sources (Ogura et al., in vivo (Meraviglia et al., In light of these considerations, the question whether special MSC sources are putatively more potent in specific disease treatments requires in depth studies, addressing comparative measures of therapeutic efficacy and safety. Several studies albeit designed and performed in a comparable fashion reported that superficially equal MSC resulted in different outcomes Once safety of MSC administration can be granted, the next step is to obtain consistent results on sustained curative benefit. Inconsistency or failure in clinical outcomes might not be solely related to MSC preparations, but also attributable to disparate therapeutic protocols. The optimal route of MSC delivery and dosage regime are still debated (Kean et al., in vitro has often been employed to deliberately grant in vivo efficacy. As the one has very little to do with the other, in particular because differentiation in vitro is performed under tightly controlled culture conditions, better methods have been conceived. In vivo, MSC differentiation or complex functions certainly depend on a plethora of parameters, which can be hardly reconstructed in vitro. Direct interactions with micro-vascular structures as well as with other cell types, such as endothelial cells appear to play stimulating and inducing roles (Chen et al., in vivo (Collins et al., Specification and definition of efficacy is another pending question to be answered. Multi-lineage differentiation in vivo functional assays also provided means and measures to simultaneously acquire long-term safety data. There is a lack of suitable long-lived models, which would allow the sensitive risk assessment of MSC or its secretome in inducing malignancy (Bruno et al., This pointed at establishing superior methods to verify the functional capacity of MSC prior clinical applications. Thus, tests in living animal were proposed to distinctly assay intrinsic capacities to differentiate into functional tissues or to exert other desired functions. Sequential heterotopic transplantation has proven to fulfill this need. Generation of heterotopic ossicles has demonstrated not only skeletogenic potentials of single clone progenitors but also self-renewal ability. It must be stressed that A major caveat in most clinical MSC applications is the lack of long-term follow ups. Such studies would reveal those parameters that are essential in determining risk and safety issues in cell therapy. Despite some recent attempts (Brizuela et al., All authors helped with drafting and revision of the article, GL completed the manuscript. All authors approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.