Fertility counselling for young women newly diagnosed with cancer is an important field of preconceptional counselling. This qualitative, small-scale study explored how young women newly diagnosed with cancer experienced specialized fertility preservation counselling and treatment in the public Danish health care system.Semi-structured, in-depth interviews were conducted with five women below 40 years recently diagnosed with cancer. All women received fertility counselling by a fertility specialist at the Fertility Clinic, University Hospital of Copenhagen, Denmark before initiation of cancer treatment. Participants were interviewed at a place chosen by them, and interviews were recorded and transcribed verbatim. Data were analysed using systematic text condensation developed by Malterud and inspired by Giorgi’s phenomenological analysis.None of the participants were aware that chemotherapy could destroy their eggs. The participants described how specialized fertility counselling and fertility preservation contributed to a belief in life after cancer, which gave them hope that they would survive their cancer disease. Further, the women described how the possibility of fertility preservation removed a huge concern and enabled them to concentrate on their cancer treatment and on getting better.Overall, the specialized fertility counselling and treatment to preserve fertility was highly valued. The women felt it gave them a choice about their future fertility. The fertility expert presented the various fertility-preserving scenarios, and the women were content that they had an actual choice. All over the industrialized world, there is a trend among both men and women to delay the birth of a first child . PreviouIn Denmark approximately 900 women below 40 years are annually diagnosed with cancer . Over thSeveral studies indicate that fertility issues are very important to younger women with cancer . StudiesFertility counselling for women with cancer is greatly needed according to earlier studies ,17,18,21The aim of this qualitative, small-scale study was to explore how young women newly diagnosed with cancer experienced specialized fertility-preserving counselling. The women had received the specialized fertility preservation counselling within a few days or a few weeks after the cancer diagnosis and before chemotherapy treatment began. In Denmark specialized fertility-preserving counselling is offered in a public health care system financed by taxes and without payment by the patients. The programme of ovarian tissue cryopreservation started in the year 1999. Since then, almost 800 patients have had tissue frozen, 53 transplantations to 41 patients have been performed, and so far the programme has resulted in 13 children .The participants were recruited between February and May 2014 at the Fertility Clinic, Rigshospitalet, University Hospital of Copenhagen, Copenhagen, Denmark. Inclusion criteria were: women between 18–39 years of age, who had recently been diagnosed with cancer, and who had recently received fertility counselling. The women should be fluent in Danish and mentally and physically well enough to participate in an interview. Five women were asked to participate in the study, and no one declined. We aimed at recruiting participants with as much variation as possible regarding marital status (married/cohabiting or single), motherhood status (having children or not), and different choices of fertility preservation treatments .Prior to the invitation to participate in this study all women had received specialized fertility preservation counselling by a fertility specialist (K.T.M.). All women had received written information about fertility preservation and individualized oral information during the counselling. All women in the study were invited to participate in the study by K.T.M. All interviews were conducted by the first author (D.H.), and interview techniques and difficulties during data collection were discussed with author L.S. D.H. is a Master student in Public Health Science and has a Bachelor degree within qualitative sociology. L.S. is a medical doctor with a PhD in qualitative health research. K.T.M. is a clinical fertility specialist. The interview took place at a location chosen by the participant: three took place in the participant’s own home, one at the participant’s workplace, and one in a café chosen by the participant. Interviews lasted between 45 and 90 min. The interviews were conducted using a semi-structured interview guide focusing on the women’s experiences with fertility counselling and fertility preservation. To gain a better and more personal insight into the field of fertility counselling, D.H. observed a fertility counselling session at the Fertility Clinic, Rigshospitalet before conducting the interviews. Interviews were recorded and transcribed verbatim, and transcripts were re-checked against the recording. Transcripts were anonymized with fictive names. The age of the participants ranged from 25 to 38 years. The participants lived in or near the capital of Copenhagen, and all but one were in a relationship. Three of the participants had one child, while two did not have any children .The interview text was analysed using systematic text condensation (STC) developed by Malterud and inspired by Giorgi’s phenomenological analysis. According to Giorgi the purpose of the phenomenological analysis is to develop knowledge of how the informants experience their world of life in a certain field, by looking for essences and essential characteristics of the phenomena under study . Like GiThe study followed the codes of ethics from the Helsinki II Declaration. According to Danish legislation, interview studies require no approval from an ethical committee.We identified three themes regarding how the participants experienced the specialized fertility preservation counselling: 1) The feeling of being unable to control anything; 2) The importance of creating opportunities; 3) The feeling of having an active choice among the different scenarios.None of the women knew that chemotherapy could destroy their eggs. One woman described how her dream about children was devastated when she was told that she could lose her eggs as a result of the treatment.Truly, a dream was shattered when they told me that my eggs; they could possibly be affected, and that we might not have any children […]. As soon as she said that I could not have children, I was not sick with cancer anymore […] it was completely gone, completely. I had left that behind me […]. (Sanne)Katrine described how she felt the cancer as something foreign that took control of her and her life. She felt sorrow in the awareness that she might never be able to have any more children.It is absolutely horrible. To lose control is, well … Actually, it is a bit strange because you take for granted that you can have children and that is not at all something you can count on. […] but that something, you know, like cancer takes the control from you and controls your life for so long […] that is absolutely horrible. (Katrine)[…] to be sad that you might lose a child you didn’t have … well, it sounds completely crazy, but that is how you feel. (Katrine)Lotte described how having children is an epoch-making life event, which the cancer can extinguish. The fertility issue was a huge issue to her when she got her cancer diagnosis. It became secondary when she started her cancer treatment, because of existential crises and fear of death.[…] Am I even going to survive all this? How can I then think of children. […] it was the fear of death that took up most of the time or all the time at that period. […] It is fine, then they are preserved [the eggs] … but it is not at all certain that I am alive in three years. (Lotte)Camilla described how she felt overwhelmed with information, so the fertility issue did not differ from the other information. Sara described that the oncologist did not talk much about fertility, when she was diagnosed with cancer, and, like Camilla, she seemed to experience the confrontation with the fertility issue as secondary.Katrine described how she experienced the fertility-preserving treatment as a positive offer. The fertility preservation was a way to create opportunities, where the cancer had set limits.I have experienced the fertility issue as something very positive. Everything else about the cancer […] it has been like limitations and the fertility has been—is possibilities […]. All the talk about it is, after all, to create possibilities for me, where all the other stuff about the chemotherapy was a huge limitation in itself. Therefore, I have experienced it as something very, very positive […]. In all the darkness, there has actually been something to light it up. That I think has been extremely positive. (Katrine)Lotte described how the fertility counselling and treatment contributed to her belief in surviving the cancer.[…] so they take the eggs out and they do all that and it costs a huge amount of money. So they must believe that I will survive this somehow—well, since they bother to do all this, right? (Lotte)Camilla described the importance in self-determination and that the fertility preservation treatment minimized her risk of menopause and infertility as side effects. This gave her a feeling of regaining some control over her body, and made her feel that she had the opportunity to attain a normal functional body again when she had recovered from the cancer.The most important thing is that we decide for ourselves. And then, I think, when you get to the other side have as few implications as possible. To enter menopause prematurely, that is a fairly bothersome implication of having had cancer, so when I hopefully get—or when I am declared well—then we should like to reach some sort of normality that things are as they have always been. (Camilla)This indicates that the women felt that fertility counselling and treatment created some new opportunities for them, when they felt unable to control anything, because of the cancer diagnosis and risk of infertility as a side effect. Lotte described how the fertility preservation treatment removed a huge concern, which she did not have to think about until after the cancer treatment. One reason given for choosing cryopreservation of ovarian tissue was to lower the risk of early menopause. Also cryopreservation of ovarian tissue made the women feel that they would attain some sort of bodily normality again.The women’s argument against receiving fertility preservation was that it meant another demand on their body, besides the cancer treatment. Sara described the fertility preservation as a substantial load, which possibly could delay her chemotherapy treatment. Lotte described how it is important to consider whether the wish to have children balances the bodily load of fertility preservation. Contrary to this, Camilla and Katrine described how they experienced the fertility preservation as a minor treatment. Even though Sara rejected fertility preservation she is grateful that she has been made aware of the fertility issue and has been offered advice on possibilities, so that she could make her own decision.Even though the women have made different decisions concerning fertility preservation, they all described gratitude for the opportunity to preserve their fertility.[…] I can say that generally I have been really pleased about it. So I just think that it is important that you […] turn the fertility thing into something positive. Not turn it into something negative and say ‘Oh no, now I can’t have children’ and then focus on that, but instead say ‘oh yes, I might have children after all’ and focus on this and get this perspective on it. (Katrine)Just the fact that there has been someone who would give me the opportunity to get to the other side and choose for myself, that has meant a lot—that has truly meant a lot. (Sanne)Well … Actually I think that, as a whole, it was fine—positive, kind, and nice people. (Sara)The women described it as a good experience and service when the various fertility preservation scenarios were presented to them. They were all pleased and comfortable with the ultrasonography of their ovaries and estimate of their ovarian reserve. They further described the importance of the fertility preservation treatment being their option.[…] she explained it all well and put up the different scenarios in a very clear way so there was still something to discuss. But it was an active choice you had to make—that I think was very important. (Camilla)The women’s considerations vary in relation to their decision about receiving fertility preservation. Regardless of their decision, several women implied that they did not know their cancer and treatment status when they had to consider the fertility preservation treatment. This complicated their decision-making process. They had to make a decision about the fertility-preserving treatment before they knew about the cancer treatment.This analysis showed how the women experienced that the fertility-preserving counselling and treatment gave them the opportunity to regain some of the lost control, by having an active choice among the different scenarios that were listed up for them at the fertility preservation counselling.In this study, it appears that fertility issues are of great importance for women below 40 years diagnosed with cancer, which is in line with results from international studies ,16–18,21Previous studies found that potential loss of fertility could have a profound influence on young women and sometimes be more stressful than the cancer itself ,18. In tA qualitative study from England  showed aA Dutch study found that women affected by cancer welcomed the ability to preserve their fertility despite the cancer diagnosis, and that the women associated the fertility counselling and treatment with positive emotions, such as hope, relief, and a reason to live . This agPrevious studies found that when women received information about possible fertility preservation treatments they felt they were given a choice on this issue and thus on their own future, which was a positive experience ,18. In tA study from the UK found that even though women were referred to a fertility specialist, it could be difficult to arrange the counselling because of long waiting times . All womGenerally, findings in this study thus appear to be consistent with the findings of previous studies in relation to young women with cancer and their experiences with fertility-preserving counselling and treatment.In this study it is a strength that all women had specialized counselling with the same professional fertility expert and that the interviewer, through observation, obtained acquaintance with the field before conducting interviews. Further, it is a strength that the participants differed with regard to marital and motherhood status and with regard to which fertility preservation treatment they chose. This we believe has implications on how they experienced the specialized fertility counselling and treatment. All participants were invited by the fertility specialist (K.T.M.) who conducted the fertility preservation counselling. On the one hand this ensured that the women were invited by a health professional they knew beforehand and, further, that women who were very fragile were not invited. On the other hand, this recruitment strategy could imply that the participants felt an obligation to be more positive towards the specialized fertility counselling than they really were. It was emphasized before the interviews were conducted that the participants should feel free to say what they wanted and that participation or not would have no impact regarding their fertility preservation treatment. The main limitation of the study is the inclusion of only five women. The point of saturation during data analyses was not reached. However, the in-depth interviews, lasting 45 to 90 min, contributed with relevant findings to previously published knowledge.In conclusion, we found that specialized fertility counselling and treatment was highly valued because the women felt they were given a choice about their future fertility. The fertility expert presented the various fertility-preserving scenarios in a satisfying way, and the women were content to have an actual choice. Further, the specialized fertility counselling contributed to a belief in own survival among the women.


The appearance of the first animal species on earth coincides with the emergence of transforming growth factor β (TGFβ) pathways. The evolution of these animals into more complex organisms coincides with a progressively increased TGFβ repertoire through gene duplications and divergence, making secreted TGFβ molecules the largest family of morphogenetic proteins in humans. It is therefore not surprising that TGFβ pathways govern numerous aspects of human biology from early embryonic development to regeneration, hematopoiesis, neurogenesis, and immunity. Such heavy reliance on these pathways is reflected in the susceptibility to minor perturbations in pathway components that can lead to dysregulated signaling and a diverse range of human pathologies such as cancer, fibrosis, and developmental disorders. Attempts to comprehensively resolve these signaling cascades are complicated by the long-recognized paradoxical role the pathway plays in cell biology. Recently, several groups have probed examples of the disparate aspects of TGFβ biology in a variety of animal models and uncovered novel context-dependent regulatory mechanisms. Here, we briefly review recent advancements and discuss their overall impact in directing future TGFβ research. Ligands of the TGFβ superfamily such as TGFβ(1–3), bone morphogenetic proteins (BMPs), activins, nodal, and growth differentiation factors (GDFs) control numerous aspects of animal biology including embryonic development, organogenesis, cell fate decisions, immune modulation, stress responses, and stem cell function7. Owing to widespread functional diversity, disruptions in TGFβ signaling are also associated with human diseases including cancer, fibrosis, systemic sclerosis, and hereditary disorders12. TGFβ morphogens often drive contradictory biological outcomes  by inhibiting Wingless-type MMTV integration site (WNT)/β-catenin signaling, yet promote specification, expansion, and homing of hematopoietic progenitors21. Although multi-layer regulation of TGFβ signaling can theoretically explain the dichotomy of activities, the exact molecular contexts that lead to opposite biological output in intact tissues remain elusive. Recent studies utilizing multiple animal and human models have addressed these contradictions and uncovered complex interactions in immune and regenerative tissues that determine specific biological outcomes of TGFβ activity. This review will briefly summarize these findings.Transforming growth factor β (TGFβ) signaling pathways have co-evolved with animalsutcomes , which h22. Activated ligands, released by the cleavage of their pro-domains in the extracellular matrix, bind to a complex of two type II and type I transmembrane receptors, inducing type II-mediated phosphorylation of type I receptors at specific serine/threonine residues. This in turn triggers the activation of regulatory Smad (R-Smad) transcription factors, which then complex with the co-transcription factor Smad4 and accumulate in the nucleus, where they alter gene expression22. Inhibitory Smads (I-Smads), which are themselves induced by TGFβ family signaling23, function in a negative feedback loop to regulate TGFβ signaling. The mammalian genome encodes 33 ligands, five type II and seven type I receptors, and eight Smad proteins . InDrosophila, there are seven ligands, three type II receptors, two type I receptors, two R-Smads , one co-Smad (Medea or Med), and one I-Smad (Dad)25. TGFβ signaling is further divided into discrete pathways, as functionally redundant TGFβ ligands generally activate a restricted set of corresponding receptors and Smad transcription factors. For example, TGFβ(1–3) activate Smad2/3 via TGFβ receptors, BMPs stimulate Smad1/5/8 activation via BMP or activin receptors, and activins, inhibins, and GDFs activate activin receptors to phosphorylate Smad2/322. However, there are exceptions to this general model, as ligand/receptor promiscuity can lead to crosstalk between these distinct pathways27.Canonical TGFβ Smad signaling is fairly simple+ crypt base columnar ISCs that reside at the bottom of crypts of Lieberkühn28. As ISCs divide asymmetrically, their progeny expands in the transit-amplifying compartment and differentiates into functional lineages as they migrate towards the tips of villi. This baso-apical structural morphology in the mammalian intestine is maintained by two opposing concentration gradients of WNT and BMP signaling pathways29 that sustain the basal regenerative stem/progenitor cell zone and apical differentiated zone, respectively. Indeed, in humans, inactivating mutations in adenomatous polyposis coli (APC) as well as the BMP receptor 1A (BMPR1A) in ISCs lead to hyperactivate β-catenin function and loss of BMP signaling, respectively, thus enhancing stem cell features and predisposing mutation carriers to cancer development31. TGFβ also promotes differentiation in the apical regions, and inactivating mutations in core TGFβ components are frequently detected in colorectal cancer (CRC). Intriguingly, recent studies propose that the activity of the canonical TGFβ pathway is enhanced by the non-canonical WNT ligand Wnt5A during acute injury in the intestine, and this promotesde novo generation and fission of crypts that facilitate efficient repair of the intestine33. At the molecular level, since the Hippo signaling pathway co-transcription factor yes-associated protein (YAP) is also required for crypt regeneration34, interaction between TGFβ and Hippo pathways may explain these observations (see below).Tissue homeostasis is maintained by a rapidly cycling population of LGR537. The activity of the immune system, however, has to be tightly controlled to prevent inflammatory disorders and maintain homeostatic regeneration of tissues. TGFβ signaling is involved in the regulation of these processes43. Previously, TGFβ signaling was shown to prevent inflammation of the bowel upon activation in the associated dendritic cells44. New data shed further light on this process by suggesting thatClostridium species within the microbiota induce TGFβ1 expression in colonic dendritic cells in a model of experimental colitis45. This in turn stimulates the generation of immunosuppressive regulatory T (Treg) cells, thereby ameliorating the bowel colitis. TGFβ1 also amplifies its own production by activating a Smad3-dependent autocrine signaling loop within dendritic cells, which is, surprisingly, inhibited by Smad2 activity45. Although Smad2 regulates TGFβ receptor expression in this context, it is not clear what drives the opposing roles of the closely related Smad3 and Smad2.The intestinal epithelium is inhabited by trillions of microbes that are kept in check by diverse epithelial and cellular immune responsesDrosophila. Macrophages are recruited to the intestinal epithelium upon injury where they secrete the TGFβ/BMP homologue Decapentaplegic (Dpp)46. Dpp then activates the type II receptor Punt (Put) and type I receptor Saxophone (Sax) complex in the ISCs, which drives nuclear localization of dSmad2, thereby triggering ISC proliferation. This immune regulation of ISC function is required for efficient repair of the damaged intestine and is paramount for survival during pathogen invasion. Interestingly, ISCs can subsequently change their response to Dpp by expressing another type I receptor, Thickveins (Tkv), which, upon activation, phosphorylates the R-Smad Mad to restore stem cell quiescence. Oncogenic transformation of ISCs induced by ectopic expression of epidermal growth factor receptor (EGFR) and Jak/STAT pathways can supersede the growth inhibitory effects of Tkv/Mad signaling and requires chronic dSmad2 activity to promote tumor-like stem cell growth and hyperplasia46  subtype was recently linked to the activation of TGFβ signaling within cancer-associated fibroblasts (CAFs), which increased the frequency of tumor-initiating cells and the promotion of metastatic invasion in humanized mouse models51. Parallel observations were also made in breast cancer patients where both heat shock factor 1 (HSF1) and TGFβ signaling activation in CAFs is associated with poor clinical outcome52. Another study showed that TGFβ signaling induces a mesenchymal transcriptional footprint in genetically engineered intestinal organoid cultures carrying a BRAFV600E mutation53  to promote oncogenic growth in KrasG12D-driven cancers. Restoration of Smad4 activity disrupts Sox4–KLF5 interaction and redirects Sox4 to induce the transcription of pro-apoptotic genes through a lethal epithelial-mesenchymal transition (EMT) program55  in the cytoplasm, which can also lead to their degradation. Upon inhibition of MST/LATS kinase cassette, YAP/TAZ translocate into the nucleus, where they associate with the DNA-binding partners, such as the TEA domain transcription factors (TEADs), to regulate gene expression. The Hippo pathway is an environmental sensor that responds to a wide range of extrinsic cues that include cell density, cell polarity, metabolic cues, GPCR signaling, and mechanotransduction. The Hippo pathway in turn interacts with several signaling cascades including TGFβ, WNT, EGFR/mitogen-associated protein kinase (MAPK), and others to regulate cell proliferation, tissue growth, apoptotic resistance, and tumor progression58. In the case of TGFβ signaling, when the Hippo pathway is inactive, YAP/TAZ physically interact with the Smad2/3–Smad4 complex to promote its nuclear accumulation, thereby regulating diverse transcriptional programs that maintain pluripotency in embryonic stem cells61, regulate differentiation into diverse lineages67, enhance wound healing68, and promote tumorigenesis in breast cancer, mesothelioma, and hepatocarcinoma models71. Recent studies have also uncovered cooperative interactions between YAP and TGFβ-regulated Smads that couple mechanotransduction in fibroblasts to TGFβ-induced profibrotic gene expression programs that are important in fibrosis73. Interestingly, the TGFβ pathway can also regulate Hippo signaling by interacting with the scaffold protein RASSF1A, which is often inactivated in human cancers74. Mechanistically, high TGFβ activity promotes ubiquitination-based degradation of RASSF1A. This inactivates the MST/LATS kinase cassette, facilitating nuclear localization of the YAP/Smad complex, thereby enhancing the tumorigenic potential of TGFβ signaling. Together, these studies indicate that the TGFβ and Hippo pathway interactions are finely tuned and disruptions in these processes can alter stem cell function and cell fate decisions and lead to fibrosis and cancer.The Hippo pathway plays a critical role in cell and organ growth and is implicated in tissue regeneration and cancer75. BMPs have been shown to specify hematopoietic tissues during development and promote heterogeneity by differentially regulating the formation of diverse hematopoietic stem cell (HSC) types that are separately deposited in the fetal liver and bone marrow of developing embryos76. Moreover, BMPs modulate myeloid lineage specification and promote homing and proliferation of hematopoietic progenitors at postnatal stages77. TGFβ(1–3) ligands, however, are potent inhibitors of hematopoiesis in adults78. New studies now identify that megakaryocytes secrete the bulk of latent TGFβ1 in the bone marrow, which is cleaved by the non-myelinating Schwann cells80. Once activated, TGFβ1 stimulates canonical Smad2/3 signaling in multipotent HSCs to promote quiescence. Intriguingly, megakaryocyte-derived TGFβ1 also protects HSCs from exhaustion in a hostile microenvironment, which could be relevant to the development of hematological malignancies79. Indeed, leukemia-initiating cells (LICs) in chronic myeloid leukemia (CML) derive therapeutic resistance from active TGFβ signaling81. Although a direct link between megakaryocytes/platelets and the development of hematological malignancies has not been established, platelets are shown to activate TGFβ signaling in circulating tumor cells, enhance immune evasion, and facilitate bone metastasis83.TGFβ pathways control several aspects of blood formation85. Among these, diffuse intrinsic pontine gliomas (DIPGs) almost exclusively occur in children at the age of 6–8 years. DIPGs exhibit a dismal prognosis with a median survival rate of only 1 year86 because no therapeutic treatment is effective against these cancers, primarily owing to their anatomical location. Whole genome exploration studies have identified unique epigenetic changes in DIPG specimens with 78% of patients exhibiting a single amino acid mutation in a histone H3 variant, a lysine replacement to methionine at position 27 (K27M). This histone H3-K27M mutation removes trimethylation from the histone H3 tail, allowing transcriptional derepression that is associated with the induction of stem cell features and cancer development87. Intriguingly, four independent studies have now shown that a majority of patients who have the K27M mutation also bear activating mutations in the activin receptor type 1 (ACVR1)92. Activity of mutant ACVR1 still requires ligand-mediated activation of type II receptors and does not seem to be tumorigenic alone, since the same ACVR1-activating mutations found in hereditary fibrodysplasia ossificans progressiva (FOP) do not result in cancer predisposition94. ACVR1 has not been previously implicated in cancer, thus the link with a mutated histone H3 variant in the context of glioma is particularly intriguing.Gliomas are the most common tumors of the central nervous systemExtensive efforts in TGFβ research have revealed that dynamic interactions between tissues and rapidly changing molecular contexts determine the specific biological output of TGFβ signaling in intact tissues. These complex processes are designed to fine-tune cellular reactions and prevent exaggerated responses that may lead to lethal consequences in animals. These interactions also allow TGFβ pathways to sense a wide spectrum of constantly changing cellular microenvironments and launch appropriate responses. This complex pattern of tight TGFβ regulation also enables just a few components to control diverse biological activities and creates opportunities for diseased cells to adapt and modify these elements to use for their own growth and survival. Therefore, while it is necessary to keep an open mind when considering such complexities in TGFβ signaling, caution must be exercised when using current tools and interpreting specific observations. This is particularly true in pathological tissues where canonical signaling pathways may be altered. In future work, it will be vital to incorporate information about TGFβ pathway crosstalk and couple observations of pathway activity to mechanistic molecular and functional assays.ACVR1, activin receptor type 1; APC, adenomatous polyposis coli; BMP, bone morphogenetic protein; CAF, cancer-associated fibroblast; CRC, colorectal cancer; DIPG, diffuse intrinsic pontine glioma; Dpp, Decapentaplegic; EGFR, epidermal growth factor receptor; GDF, growth differentiation factors; HSC, hematopoietic stem cell; ISC, intestinal stem cell; I-Smad, inhibitory Smad; K27M, lysine replacement to methionine at position 27; Klf5, Kruppel like factor 5; R-Smad, regulatory Smad; Sax, Saxophone; TAZ, transcriptional co-activator with PDZ-binding motif; TGFβ, transforming growth factor beta; Tkv, Thickveins; WNT, Wingless-type MMTV integration site; YAP, yes-associated protein.


Saccharomyces cerevisiae. In addition to the well-known specific responses to starvation for an essential nutrient, like nitrogen or phosphate, the presence of fermentable sugar or a respirative carbon source leads to predominance of fermentation or respiration, respectively. Fermenting and respiring cells also show strong differences in other properties, like storage carbohydrate levels, general stress tolerance and cellular growth rate. However, the main glucose repression pathway, which controls the switch between respiration and fermentation, is not involved in control of these properties. They are controlled by the protein kinase A (PKA) pathway. Addition of glucose to respiring yeast cells triggers cAMP synthesis, activation of PKA and rapid modification of its targets, like storage carbohydrate levels, general stress tolerance and growth rate. However, starvation of fermenting cells in a glucose medium for any essential macro- or micro-nutrient counteracts this effect, leading to downregulation of PKA and its targets concomitant with growth arrest and entrance into G0. Re-addition of the lacking nutrient triggers rapid activation of the PKA pathway, without involvement of cAMP as second messenger. Investigation of the sensing mechanism has revealed that the specific high-affinity nutrient transporter(s) induced during starvation function as transporter-receptors or transceptors for rapid activation of PKA upon re-addition of the missing substrate. In this way, transceptors have been identified for amino acids, ammonium, phosphate, sulfate, iron, and zinc. We propose a hypothesis for regulation of PKA activity by nutrient transceptors to serve as a conceptual framework for future experimentation. Many properties of transceptors appear to be similar to those of classical receptors and nutrient transceptors may constitute intermediate forms in the development of receptors from nutrient transporters during evolution. The nutrient-sensing transceptor system in yeast for activation of the PKA pathway has served as a paradigm for similar studies on candidate nutrient transceptors in other eukaryotes and we succinctly discuss the many examples of transceptors that have already been documented in other yeast species, filamentous fungi, plants, and animals, including the examples in human cells.The nutrient composition of the medium has dramatic effects on many cellular properties in the yeast  Saccharomyces cerevisiae growing in a complete nutrient medium with a fermentable carbon source display a range of phenotypes indicating that the activity of PKA is high, while the opposite is true for cells growing on a non-fermentable carbon source or for stationary phase cells  cells in which the cAMP-PKA pathway had first been activated by glucose, as evidenced by transient activation of trehalase, and to which a nitrogen source was added to support growth initiation by the cells. Interestingly, addition of the nitrogen source caused an immediate reactivation of trehalase, without any change in the cAMP level . This in0 . As a reBy using temperature-sensitive mutants in cAMP synthesis at permissive and restrictive temperature it was confirmed that nitrogen-, phosphate-, and sulfate-induced activation of trehalase do not use cAMP as second messenger, although maintenance of a basal level of cAMP is required for full activation . BecauseS. cerevisiae will be discussed in the next section. An overview of the different nutrient transceptors identified in S. cerevisiae and their role in activation of PKA in concert with the cAMP-PKA pathway is shown in Figure 1.Recently, the concept of non-cAMP mediated activation of PKA through the FGM pathway has been expanded from the macro-nutrients, nitrogen, phosphate, and sulfate, to the micro-nutrients iron and zinc . In eachProving an additional nutrient sensing property of a transporter is not straightforward and requires several levels of evidence (1) excluding the involvement of intracellular sensing and thus also of other transporters that can take up the relevant substrate, (2) identification of non-transported competitive or non-competitive inhibitors with agonist signaling function, (3) transport-deficient versions of the transceptor that have retained the signaling function, for instance by mutating the residues involved in proton-coupling required for symport of the substrate , and (4)GAP1 is strongly enhanced and the protein is present in high quantity at the plasma membrane  analysis showed that transport and signaling use the same substrate binding site in Gap1  which did not trigger signaling and showed that other transported amino acids do not trigger signaling in concentrations below their Km , a G-protein that plays an essential role in the initiation of protein synthesis and cell growth . The clo unclear .As previously explained, re-addition of the nutrient for which the cells have been starved triggers transceptor endocytosis and degradation in the vacuole. Hence, transceptor signaling to PKA has to be transient. In spite of this, the cells maintain a high PKA phenotype throughout exponential fermentable growth, conditions under which all starvation-induced transceptors remain repressed. This may suggest that fermentable sugar like glucose continuously activates PKA through the cAMP pathway during fermentative growth and that starvation-induced downregulation of PKA is caused in a cAMP-independent way by the transceptors. One straightforward and plausible hypothesis could be that the free catalytic subunits of PKA bind to the transceptor(s) and remain in this way sequestrated from their targets as long as the cells remain starved for at least one essential nutrient. This would result in the typical downregulation of the whole range of PKA targets, which is observed in all stationary-phase cells. In this case, the many different transceptors would only need a common intracellular binding site for the catalytic subunits of PKA, which would be exposed when the substrate of the transceptor is absent. When the substrate is added and transported by the transceptor, the latter changes its conformation and releases the catalytic subunits of PKA, triggering the typical sudden burst in PKA activity. This hypothesis can explain in a straightforward manner why so many different nutrients can trigger the same downregulation and re-activation of PKA through their own specific transceptor using basically the same mechanism. The glucose requirement of nutrient transceptor signaling can then be explained by the requirement of maintaining an adequate cAMP level and thus also an adequate amount of free catalytic subunits of PKA. Moreover, the presence of glucose also causes inactivation of the kelch-repeat proteins Krh1,2, which strengthen the binding of the catalytic and regulatory subunits. Their inactivation thus increases the amount of free catalytic subunits even in the presence of a constant cAMP level . In the In virtually all eukaryotic cell types, transporters and receptors are found in the plasma membrane. It seems plausible that nutrient transporters originated first in evolution since for any unicellular organism nutrient import is essential for survival and proliferation. Sensing and signaling by receptor proteins appears as a more sophisticated property generated later in evolution. It is unclear how receptors originated in evolution, but the close similarity between neurotransmitters and nutrients has led many years ago already to the suggestion that receptors might originate from nutrient transporters . The latFigure 2). This could provide an evolutionary advantage to a microorganism in allowing it to adapt its metabolism, gene expression machinery and/or other cellular properties more rapidly, i.e., before the nutrient has been able to affect these properties through its metabolism, i.e., by increasing energy levels, reducing power or availability of precursors. Nutrient transporters undergo multiple conformational changes during the transport cycle. Hence, it is not difficult to envisage that a specific conformation triggered by transport of the substrate would allow the transporter to interact with an intracellular protein, whereas the transporter in its basic conformation would not be able to do so. The yeast high-affinity transporters that are strongly induced during deprivation of their substrate, Gap1, Mep2, Pho84, Sul1,2, Ftr1, and Zrt1, and that all signal to the PKA pathway, are prominent examples of such transporters that gained an additional signaling function to rapidly affect a pleiotropic range of cellular properties upon detection of their nutrient substrate. It appears likely that loss of the transport function in such transporter-receptors has led to the transporter-like proteins with a pure nutrient sensing function . These proteins have retained the capacity to bind the nutrient substrate, possibly allow it to enter to some extent the transportation channel, and as a result switch the transporter conformation into the signaling conformation. Further transport of the nutrient toward release into the cytosol would have become defunct. The yeast Snf3, Rgt2, and Ssy1 proteins are prominent examples of such nutrient sensors. The demonstration that the mammalian SGLT3 non-transporting glucose transceptor can be converted into an actively transporting carrier by a single amino acid change in the glucose-binding domain provides strong support that the transition from transporting transceptor to non-transporting receptor would be a simple evolutionary step .We may consider all transporter-related proteins with a nutrient signaling function as transporter-receptors or transceptors. Hence, there are transporting and non-transporting transceptors. The distinction between the two categories may not be sharp. It is quite likely that eukaryotic transceptors with a primary signaling function may be discovered that have retained residual transport activity. Actually, such a protein has been discovered already in activity . Intermeactivity . Conversactivity . These dThe similarity of transporting transceptors and classical receptors in undergoing substrate/ligand-induced internalization also extends to the non-transporting transceptors. The low-affinity glucose sensor Rgt2 remains stable only in high-glucose grown cells, and the high-affinity glucose sensor Snf3 is stable only in cells grown in media with a low glucose level. Both proteins undergo ubiquitination-induced internalization and degradation in the vacuole under adverse conditions. In addition, constitutively active, signaling forms of the glucose sensors do not undergo endocytosis, whereas signaling defective sensors are constitutively targeted for degradation, suggesting that the stability of the glucose sensors may be associated with their ability to sense glucose .Subsequent to the discovery of transporting and non-transporting nutrient transceptors in yeast, many examples of nutrient transceptors have been found or suggested in other eukaryotes. As in yeast, most of these transceptors have been discovered in connection with regulatory responses to deprivation or to resupplementation of the substrate of the transceptor. Hence, these transceptors generally function in specific processes controlled by the substrate of the transceptor and often related more or less directly to the metabolism and/or the uptake of the substrate. For several of these transceptors there is evidence for control of multiple downstream processes and in some cases it has been shown that separate signaling events are involved. There are no clear examples yet in other eukaryotes of transceptors for different nutrients that control the same signaling pathway, as in the case of the yeast transceptors that trigger activation of the PKA pathway. In general, disconnection between uptake activity of a transporter and/or metabolism of its nutrient substrate and one or more regulatory responses triggered by the nutrient has led to awareness that the transporter might act as a sensor. Convincing experimental demonstration generally requires further detailed genetic modification of the transporter to separate transport and sensing more clearly to show that it truly functions as a transceptor.S. cerevisiae, the Mep family members Ump2 in Ustilago maydis , was shown to control growth via a mechanism that does not require bulk transport of amino acids into the cell  by direct interaction with proline-rich tyrosine kinase-2 (PYK2) . Loss ofExpression of a transport deficient mutant form of the human CNT1 nucleoside transporter in pancreatic cancer cells affected multiple processes in a similar way as expression of the wild type CNT1 transporter, which led to the suggestion that it may function as a transceptor .D-aspartate and small molecule inhibitors, that elicited the same response as glutamate  by nutrient transceptors, in particular the signaling proteins that directly interact with the nutrient transceptors, the identification of other examples of the same cellular process affected by multiple transceptors for different nutrients, the identification of similar signaling domains and downstream signaling pathways in transceptors from multiple distantly related species and elucidation of the connection between substrate binding, transport and signaling in a transceptor.All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.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.


Diacylglycerol kinases (DGKs) terminate diacylglycerol (DAG) signaling and promote phosphatidic acid (PA) production. Isoform specific regulation of DGKs activity and localization allows DGKs to shape the DAG and PA gradients. The capacity of DGKs to constrain the areas of DAG signaling is exemplified by their role in defining the contact interface between T cells and antigen presenting cells: the immune synapse. Upon T cell receptor engagement, both DGK α and ζ metabolize DAG at the immune synapse thus constraining DAG signaling. Interestingly, their activity and localization are not fully redundant because DGKζ activity metabolizes the bulk of DAG in the cell, whereas DGKα limits the DAG signaling area localizing specifically at the periphery of the immune synapse. When DGKs terminate DAG signaling, the local PA production defines a new signaling domain, where PA recruits and activates a second wave of effector proteins. The best-characterized example is the role of DGKs in protrusion elongation and cell migration. Indeed, upon growth factor stimulation, several DGK isoforms, such as α, ζ, and γ, are recruited and activated at the plasma membrane. Here, local PA production controls cell migration by finely modulating cytoskeletal remodeling and integrin recycling. Interestingly, DGK-produced PA also controls the localization and activity of key players in cell polarity such as aPKC, Par3, and integrin β1. Thus, T cell polarization and directional migration may be just two instances of the general contribution of DGKs to the definition of cell polarity by local specification of membrane identity signaling. The compartmentalization of plasma membrane proteins is a common characteristic of eukaryotic cells and provides the base for the establishment of signaling domains    to cell organization has been extensively characterized; specific PIs play a major role in determining the subcellular identity of membranes  second messengers that recruit a set of interacting proteins  is a specialized structure described as the immunological synapse (IS)  microclusters form at the IS periphery and move toward the cSMAC where they encounter the endocytic sorting machinery and are internalized. IS formation drives the polarization of the entire T cell, with the translocation of the microtubule organizing center (MTOC) between the IS and the nucleus, and the establishment of the uropod, a membrane zone enriched in signaling molecules at the opposite end of the T cell , but other investigators have demonstrated that DGKζ plays an equivalent role in cell migration. DGKζIn fibroblasts, DGKγ is also recruited to ruffles and lamellipodia where it co-localizes with Rac1 Figure . Surpris3,4,5P3 signaling and DGKζ-produced PA , AIRC  (IG13524) and PRIN (2015 4CWJH4). GB is supported by the TIPSO grant from Regione Piemonte . Salary of VM and VB is provided by Telethon and TIPSO to AG and GB respectively.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.


Amazon Mechanical Turk (MTurk) is becoming a prevalent source of quick and cost effective data for organizational research, but there are questions about the appropriateness of the platform for organizational research. To answer these questions, we conducted an integrative review based on 75 papers evaluating the MTurk platform and 250 MTurk samples used in organizational research. This integrative review provides four contributions: (1) we analyze the trends associated with the use of MTurk samples in organizational research; (2) we develop a systems perspective  to synthesize and organize the key factors influencing data collected on MTurk that may affect generalizability and data quality; (3) within each factor, we also use available MTurk samples from the organizational literature to analyze key issues ; and (4) based on our review, we provide specific recommendations and a checklist for data reporting in order to improve data transparency and enable further research on this issue. The recent past has seen a steady increase in the number of published studies utilizing Amazon's Mechanical Turk (MTurk) platform for crowdsourcing participants and workers  compared to just seven published studies in the same journals in 2012. This trend suggests that within the next few years we will see hundreds of studies within organizational journals that utilize MTurk samples. Despite the growing use of MTurk samples within organizational and psychological research, some scholars have raised concerns about its use and current debates surround whether MTurk is appropriate for organizational research . Specifically, there is a need to address whether the various concerns often raised by editors, reviewers, and researchers should outweigh the convenience and reduced cost  to synthesize and organize past research. Using this framework, researchers can understand how specific research choices made within these systems influence the generalizability and quality of data collected on MTurk. We propose that the broader systems perspective taken here can be applied to understand key issues in other types of samples and data collection . Third, within this framework, we use available MTurk studies from the organizational literature to analyze key issues . Therefore, this review empirically establishes answers to important questions. Fourth, based on our review, we present best practices and recommendations for researchers and include a detailed checklist for MTurk data collection reporting to increase data transparency.Amazon launched MTurk in 2005 for internal work that required human intelligence. Since then, MTurk has grown immensely and is open to anyone who registers for an Amazon account. Individuals may utilize MTurk as a “worker”  or a “requestor” . Each worker is identified with a unique worker ID, which remains constant unless the worker creates a new account.A requestor can post a HIT using the internal MTurk survey platform or by posting a link to an external survey platform . A HIT may be a survey, a coding task, or any other task that requires human intelligence. Requestors must also decide upon a title, description, and keywords to describe their HIT, the payment per HIT, the number of workers that may complete a HIT, the time allotted for each HIT, the expiration date for the HIT, and the worker requirements . The HIT can be released in batches, which allows for an easy test run before collecting a large amount of data. That is, a requestor can release a small batch of HITs to obtain feedback on the HIT or be sure the data collection is running smoothly.When searching for HITs, Amazon defaults to show the newest HITs; however, workers may also sort the HITs by the oldest HITs, fewest (or most) HITs available, payment, expiration data, title (A–Z or Z–A), and time allotted. Workers are able to search for particular keywords or qualifications and specify a minimum amount of payment. Past research has suggested that workers tend to search for HITs based on which are the most recent (the default) and by the number of HITs available within a task , which permits greater researcher control and flexibility without requiring extensive programming knowledge. To use TurkPrime, a requestor simply links their created TurkPrime account to an existing MTurk account. We view TurkPrime as a valuable resource for researchers and integrate its uses into our review where appropriate.More recently, third party platforms  have emerged to make MTurk more “researcher friendly.” The most comprehensive platform as of 2017 is TurkPrime  offering reviews or tutorials of the MTurk platform, (2) examining the quality of data collected on MTurk, or (3) examining various characteristics of MTurk samples, particularly in relation to other samples. Only articles focusing on MTurk for use by researchers were considered in the current review; that is, articles not directly examining the MTurk platform and participants were excluded from the current review. Based on these criteria, we found 75 relevant articles.Academy of Management Journal (AMJ), Administrative Science Quarterly (ASQ), Journal of Applied Psychology (JAP), Journal of Business and Psychology (JBP), Journal of Management (JOM), Journal of Occupational and Organizational Psychology (JOOP), Journal of Organizational Behavior (JOB), Journal of Vocational Behavior (JVB), Leadership Quarterly (LQ), Organizational Behavior and Human Decision Processes (OBHDP), Organizational Research Methods (ORM), Organization Science (OS), and Personnel Psychology (PPsych). These journals represent top journals in the field of organization science . Approximately half of these studies were MTurk samples . That is, on average, articles containing at least one MTurk sample had around four samples in total and approximately two MTurk samples. Only four articles in the surveyed articles were single study papers  of the articles included in these analyses were multiple study papers  studies per article. Furthermore, ~40% of the articles we reviewed for the purposes of this paper from OBHDP contained five or more studies in the article (16.7% were seven or eight study articles). This finding can be compared to JAP, which had the second highest frequency of MTurk samples. Approximately 20% of the JAP articles containing at least one MTurk sample had five or six studies total; however, none had more than six studies. This suggests that MTurk may be considered a viable option for organizational researchers who wish to replicate findings with a different sample quickly.Third, out of the 138 articles reviewed, 66 (47.83%) were published in Based on the literature, we organized the research into three broad systems that examine MTurk through an organizational science lens. We argue that MTurk  can be viewed as a labor market that has an extant labor force from which to recruit, select, and manage work. Specifically, we examine MTurk as a recruitment system, a selection system, and a work management system. Each of these systems is made up of a unique set of factors that influence generalizability and data quality. We examine generalizability and data quality in particular because these criteria are highly relevant to our ability to draw inferences as researchers. For MTurk to be a viable option for organizational researchers, the sample should be representative of the target population  to nationally representative samples , MTurk samples are younger, more educated, and lower on the income scale. This demonstrates that MTurk samples are not representative of the United States. However, this is not a surprising finding given that college, Internet, and community samples are not expected to be representative of the United States.Table not representations of the general population. We should carefully consider whether representativeness of national data should be the criterion, as organizational researchers have traditionally collected data from class settings , specific organizations/communities, and even other online panel data from survey companies where participant solicitation processes are not transparent. Within organizational research, it may be more important to ask whether MTurk demographic characteristics are appropriate for the specific study goals at hand  and found that more than 20% of the 88 studies had a dropout rate greater than 30%. In contrast, the group also collected 82 student (lab) samples  finding that 96% had no attrition. However, the key issue may be less so the differences in sample but the mode of data collection. Behrend et al. . Within the United States, they are not representative of the general U.S. population and a large proportion of MTurkers reported being unemployed, are likely underemployed, and have higher negative emotions and attitudes. Therefore, when conducting a study using MTurk, it is important for organizational researchers to determine if the general MTurk population is appropriate. It is likely that selection and screening procedures will need to be implemented to obtain the appropriate participants (see subsequent sections). In general, there does not appear to be strong evidence that MTurk participants produce lower quality data. However, researchers should be aware of the dropout rates (20–30%) and need to account for this in their budgeting and research designs . In addition, researchers need to be aware that the large portion of non-English speakers in the MTurk pool that may potentially affect results .https://requester.mturk.com/tour). However, registered workers may not equate to active workers, and active workers may not equate to survey takers. There is some concern that “professional workers” or “Super Turkers” complete the majority of the organizational/psychological studies on MTurk , compared to U.S. workers who do not view MTurk as their primary job . These results demonstrate that MTurk workers can view MTurk as a job, and the workers viewing MTurk as a primary job are more likely to be represented in surveys.The presence of Super Turkers suggests that active survey workers may view MTurk as a full-time job or may view their full-time work in significantly different ways from workers who do not actively engage in paid survey responding. For example, Brawley and Pury  found thper se. However, repeated participation in similar experiments may pose a problem for the accuracy of conclusions drawn from MTurk samples due to exposure effects. To illustrate, 59 and 52% of workers self-reported exposure to the prisoner's dilemma and ultimatum paradigms, respectively  by other requestors. Researchers can specify the range of approval percentage desired ranging from 0 to 100. Researchers can also specify the number of HITs approved, which is essentially a qualification that stipulates the level of experience on MTurk. Finally, researchers can specify the location of the workers to be of a particular country or even a specific state(s) within the United States. In addition to the standard qualifications, custom qualifications can also be made.n = 130). Out of these samples, 103 (79.2%) specified U.S. workers, 10 (7.7%) specified an approval rating , and 63 (48.5%) created a custom qualification . Of the samples using qualifications, the majority reported using one (n = 92 or 70.77%) or two (n = 30 or 23.08%) qualifications.How are qualifications used and reported within organizational research? We found a lack of homogeneity in whether qualifications were used and the number of qualifications used. Out of 250 samples used in organizational research, about half of the samples (52%) reported the use of qualifications  would limit generalizability to a broader demographic; however, this may increase validity if the research question is relevant to the demographic sampled .The choice of standard qualifications can enhance data quality. Foremost, for higher quality data, we recommend specifying a 95% approval rating or above to increase the chances of higher quality data  rather than custom qualifications for selecting particular worker characteristics.Prescreens are HITs  used to indirectly screen out workers who are not in the desired target population  or that do not exert sufficient effort . Prescreens have be used to select workers with specific work experiences  or be more likely to participate in certain types of studies  that may indicate the extent of self-selection into and out of HITs , work design , and employing attention checks. Uncontrollable factors include the presence of MTurk message boards where workers discuss HITs.Prior to launching a HIT, researchers set a payment amount for a completed HIT. Work done on MTurk must be either automatically or manually approved by the researcher for the workers to be compensated. Generally, work completion is verified with a code found at the end of the study.There are two types of compensation that may be awarded through Amazon: base pay and bonuses. Bonuses are optional, but can be awarded as an extra incentive, such as for higher quality work . Assuming roughly 30-min studies, this appears consistent with past research suggesting a payment of $0.75 for a 30-min survey  reported a compensation amount. Compensation ranged from $0.10 to $5.00, with an average compensation amount of about $0.99 , followed by judgment and decision-making topics (n = 30) and ethical, legal, diversity, and international issues (n = 26). The high representation of these three topics should not be surprising. As previously stated, around 20.8% of the MTurk samples used a correlational design making attitudinal research a popular option on MTurk. Likewise, another 69.2% of the samples used an experimental design making judgment and decision-making research a viable option. Finally, MTurk samples are more diverse than other samples creating opportunity for researchers interested in cultural issues or diversity more generally.In our survey of the organizational literature utilizing MTurk, the experimental manipulations used on MTurk ranged broadly from avatar video-based studies  designs. We discuss how research designs may be related to the issues of generalizability and data quality.There has been little research examining how the different types of research designs may impact generalizability given the types of MTurk workers who may be most attracted to such studies. However, there is some indirect evidence that can be gleaned on this issue depending on the type of research design. We have discussed issues pertaining to non-experimental cross-sectional research in previous sections, thus we focus on experimental and longitudinal designs here. With regard to experimental designs, past research examining the MTurk platform has generally replicated various common experimental paradigms and effects   rates are higher. According to the MTurk literature   participants who have participated in similar experiments . Out of the samples reporting an attention check, the majority used direct techniques for detecting inattention: 25 (43.9%) included at least one instructed item, 1 (1.8%) included at least one bogus item, 9 (15.8%) asked the worker to recall information from instructions or an article, and 1 (1.8%) directly asked workers if they were paying attention. 12.28% of the attention checks were archival: 6 (10.53%) monitored the time spent on the task and 1 (1.8%) examined the data for long-string or invariant responding. The remaining 17 samples had unspecified attention checks. Thus, no samples reported using statistical methods for detecting insufficient effort responding. Only two samples  being excluded, which equates to roughly 13% of the sample, on average.We examined the MTurk samples in the organizational journals surveyed and found that only 22.8% of samples reported the use of attention checks  that may contain information about the nature of the HIT, the presence and nature of data-quality check questions, payment, and so forth . Another use of monitoring message boards is to track if negative information about the study has been put online. This could substantially lower the response rates on a research HIT. In addition, if researchers are concerned that compromising information may be shared on the message boards eventually, they should consider releasing the research HIT in one large batch to alleviate time for online discussion , a selection system (factors involved in selecting participants), and a work management system (factors involved in monitoring and managing the research). The individual factors comprising these systems will vary; however, the overarching systems remain the same. We propose that researchers can consider other types of data collection modes  within this frame of reference to consider the appropriateness of a given sample for the research questions at hand.As we note in many sections, there is a lack of research examining how various factors influence generalizability and data quality. We urge researchers to consider the areas noted in Supplementary Tables The aim of the current article was to provide organizational researchers with an integrative resource with which to judge the appropriateness of MTurk samples for use in their research. We organized past research examining and utilizing MTurk into an organizing framework that views MTurk through a systems perspective. Our general conclusion is that MTurk provides researchers with a unique opportunity to sample a wider portion of the working population than traditional student or community samples and in a quick and cost effective way. With opportunities, however, come challenges unique to MTurk. We view MTurk as another means of data collection and we urge researchers to consider their research design and research questions. We hope that this article presents the information necessary for organizational researchers (and researchers in related fields) to make educated decisions about whether or not to utilize MTurk and how best to do so. We additionally hope that the current review will stimulate future research to fill current gaps in our understanding of how MTurk impacts our science.MK: Wrote the paper, conducted literature search, and coded the studies when necessary. LT and PH: Helped with the framing of the paper, writing, and editing.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.


An increased risk of total death owing to human T-lymphotropic virus type-I (HTLV-I) infection has been reported. However, its etiology and protective factors are unclear. Various studies reported fluctuations in immune-inflammatory status among HTLV-I carriers. We conducted a matched cohort study among the general population in an HTLV-I-endemic region of Japan to investigate the interaction between inflammatory gene polymorphisms and HTLV-I infection for total death, incidence of cancer, and atherosclerosis-related diseases.TNF-α, IL-10, and NF-κB1. A Cox proportional hazard model was used to estimate the hazard ratio (HR) and the interaction between gene polymorphisms and HTLV-I for risk of total death and incidence of cancer and atherosclerosis-related diseases.We selected 2180 sub-cohort subjects aged 35–69 years from the cohort population, after matching for age, sex, and region with HTLV-I seropositives. They were followed up for a maximum of 10 years. Inflammatory gene polymorphisms were selected from TNF-α 1031T/C and HTLV-I for atherosclerosis-related disease incidence was statistically significant (p = 0.020). No significant interaction was observed between IL-10 819T/C or NF-κB1 94ATTG ins/del and HTLV-I. An increased HR for total death was observed in the Amami island region, after adjustment of various factors with gene polymorphisms .HTLV-I seropositivity rate was 6.4% in the cohort population. The interaction between TNF-α 1031T/C and HTLV-I to be a risk factor for atherosclerosis-related disease. Further follow-up is warranted to investigate protective factors against developing diseases among susceptible HTLV-I carriers.The present study found the interaction between  •We observed death and incidence risk with HTLV-I and SNPs in a matched cohort study.•An increased risk for total death with HTLV-I was observed in the island region.•Inflammatory SNP interacted with HTLV-I for atherosclerosis-related disease risk. Increased production of T-helper (Th)-1 pro-inflammatory cytokines  study, which has been described elsewhere.We followed up with all subjects to account for movement out of the study region or death using local government records until the end of 2015, with the exception of subjects from one mainland region, who were followed up until 2013. This follow-up in the total cohort revealed 238 cases of movement out of the study region and 164 deaths. There were 29 deaths in the sub-cohort subjects.Information regarding the incidence of cancer, myocardial infarction, aortic aneurysm, and stroke was collected using direct interview at 5–6 and 10 years after the baseline. For subjects whom we were unable to interview, information was collected via mail. The responses to the question on incidence in both the interview and the questionnaire always included “yes”, as well as “not suffered”. Causes of death were collected using death certificate records from the public health center, and deceased subjects were included in incident cases. The information of incident cases was not yet obtained for the mainland region, because the baseline survey was conducted in 2012 and the follow-up survey for incident cases of the mainland is scheduled in 2017. The coverage rate of the incident survey was 81.3% in the Amami island regions. The numbers of incident cases of cancer, myocardial infarction, aortic aneurysm, and stroke were 68, 17, 2, and 14, respectively. The survey to the medical clinics/hospitals to confirm the diagnosis for screened cases is currently ongoing. We partially evaluated the positive predictive value for confirmed diagnosis in screened incident cases of cancer and atherosclerosis-related disease among those whom we could survey by May, 2016. The number of confirmed cases using medical records in the medical clinics/hospitals was 22 of 23 screened cases (95.6%) for cancer incidence and 15 of 16 (93.8%) screened cases for atherosclerosis-related disease incidence.We used a J-MICC standardized structured questionnaire to collect individual information about socio-demographic characteristics, smoking, alcohol consumption, exercise, dietary habits, personal and family medical history, intake of prescription medicines and supplements, reproductive history, and stress status.Health checkup data obtained from the Kagoshima Kouseiren Health Center and Japan Health Club, included systolic and diastolic blood pressure, levels of total cholesterol, triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and fasting blood glucose (FBG). The subjects were asked to fast for >10 h. LDL-C levels were calculated according to the Friedewald formula using a TG level <400 mg/dLCAVI using Vasera VS-1000 and VS-1500 vascular screening systems  was used to evaluate arterial stiffness.22Plasma samples were obtained at the baseline and were stored at −80 °C prior to the examination. We screened for HTLV-I antibody using a particle agglutination assay and confirmed using western blot analysis at the laboratory of the Fukuoka Red Cross Center and SRL Inc. Subjects with indeterminate western blot results were excluded from the study.DNA was extracted from the buffy coat fraction using the standard method and the QIAamp Blood Mini Kit , GenElute Blood Genomic DNA Kit , or Blood-Animal-Plant DNA Preparation Kit .TNF-α 1031T/C [rs1799964], IL-10 819T/C [rs1800871], and NF-κB1 94ATTG ins/del [rs28362491].2O. The cycling was initiated by heating at 95 °C for 10 min, followed by 50 cycles each at 95 °C for 15 s, 58 °C for 1 min, and 25 °C for 30 s.We selected three single-nucleotide polymorphisms (SNPs) with minor allele frequencies (MAF) greater than 0.05 from the common genes for cytokines that have important roles in the immune-inflammatory processes of HTLV-I infection, cancers, and atherosclerosis, including el rs283691.24, 25TNF-α 1031T/C ([CC & CT] and TT), IL-10 819T/C ([CC & CT] and TT), and NF-κB1 94ATTG (ins/ins and [ins/del & del/del]).Hypertension was defined based on a systolic blood pressure ≥140 mm Hg, diastolic blood pressure ≥90 mm Hg, or treatment with antihypertensive medication. Dyslipidemia was defined based on TG levels ≥150 mm/dL, LDL-C levels ≥140 mg/dL, HDL-C levels <40 mg/dL, or the use of antihyperlipidemic medication. Glucose intolerance was defined as FBG value ≥110 mg or those who were being administered diabetes mellitus treatment. Smokers were defined as current smokers, and ≥20 pack-years. Alcohol consumers were defined as those who currently consumed alcohol three times or more per week. Selected food and beverage intake was categorized according to an intake frequency of three times or more per week. Metabolic equivalents (METs) were calculated based on the frequency, intensity, and duration of habitual exercise and daily activities , as obtained from the questionnaire. Estimation of METs per day was based on duration and the intensity of the exercise, using values of 3.0 for walking, 3.3 for light exercise, 4.0 for moderate exercise, 4.5 for heavy work, and 8.0 for heavy exercise.The participants' background characteristics, atherosclerosis-related factors, current and past medical history, and the genotype distribution of SNPs were compared according to HTLV-I seropositivity by sex, and differences in the prevalence were evaluated using the chi-square test. The differences in genotype distribution from the Hardy–Weinberg equilibrium (HWE) were evaluated using Pearson's chi-square test.p-value of <0.05 were considered statistically significant.Person-years were calculated from the date of the baseline until the date of death, movement out of the study area, incidence of cancer and atherosclerosis-related diseases, or the end of the follow-up, whichever came first. The date of the end of the follow-up was set as December 31, 2015 for death and movement out of the study area, except for one mainland region, for which it was set as December 31, 2013, and the most recent surveyed date was used for incidence. A Cox proportional hazard model was used to estimate the hazard ratios (HRs) and its 95% confidential intervals (CIs) for death and incidence with HTLV-I positivity, after adjustments for smoking, drinking, meat intake, hypertension, dyslipidemia, glucose intolerance, family history of coronary heart disease (CHD) and stroke, overweight, and exercise habit. The incident cases of myocardial infarction, aortic aneurysm, and stroke were combined as atherosclerosis-related diseases, because of the limited numbers of events. The interaction between each gene polymorphism and HTLV-I for the HR of death and incidence with HTLV-I was tested, after adjustment for related factors. All statistical analyses were performed using Stata software , and differences with a p < 0.001). The seropositivity rate differed by region, with mainland subjects of both sexes showing a higher rate than Amami island subjects (9.2% vs. 3.8% in men and 9.9% vs. 5.5% in women) (data not shown).The mean age of the total cohort subjects was 57.3  years for men and 57.5  years for women. The HTLV-I seropositivity rate was 6.4% (464/7210 subjects), with women showing a higher rate than men (7.0% vs. 5.6%). The seropositivity rate increased with age in both sexes ; however, the NF-κB1 94ATTG ins/del polymorphism was not (p = 0.013). The MAF of the TNF-α 1031T/C  and IL-10 819T/C  were different by region, but that of the NF-κB1 94ATTG ins/del was similar  (data not shown).The MAF of the TNF-α 1031T/C for total death and incidence of cancer and atherosclerosis-related diseases with HTLV-I seem to be higher than those of the TT genotype, and the interaction between TNF-α 1031T/C polymorphism and HTLV-I for incidence of atherosclerosis-related diseases was statistically significant  and after adjustment for additional cancer and atherosclerosis related factors , but evaluation was difficult in the mainland due to small number of person-years . The incIn the present matched cohort study, we observed a positive association between HTLV-I infection and the risk of total death in the Amami island region, which is in accordance with previous reports of increased risk of mortality for all causes other than ATLL and non-neoplastic diseases.Geographical variation in SNP distribution among Japanese has been reported, and most Japanese individuals fell into two main clusters, Hondo from the main islands in Japan and Ryukyu from Okinawa.α, encoded by the TNF-α gene, is a cytokine responsible for resistance to infection and cancers and exerts major proinflammatory action by stimulating adhesion molecules and chemokine expression.TNF-α 1031T/C is functional in increasing gene transcription and cytokine levels.TNF-α 1031T/C polymorphism and cancer risk is controversial. Allele C was associated with an increased risk of gastric cancerα is involved in lipid metabolism, coagulation, insulin resistance, and endothelial function, while TNF-α 1031 allele C was associated with CHD risk and increased dyslipidemia.TNF-α 1031T/C polymorphism and HTLV-I with an increased risk of atherosclerosis-related disease incidence. The HRs for atherosclerosis-related disease incidence with HTLV-I in CC and CT genotypes were consistently higher than those in the TT genotype, including in the case of total death and cancer incidence, although the HRs were not statistically significant.The associations between inflammatory gene polymorphisms and the development of ATLLIL-10 595A/C polymorphism showed a higher prevalence in HAM/TSP patients with higher HTLV-I provirus load.IL-10 819T/C polymorphism was associated with increased risk for gastric cancer.IL-10 819T/C polymorphism and HTLV-I for the risk of mortality and incidence with HTLV-I.IL-10 is a regulatory cytokine responsible for counterbalancing the inflammatory effects of pro-inflammatory cytokines, and its levels are decreased in HTLV-I-associated inflammatory diseases.κB1 is an important transcription factor modulated by Tax, which plays a critical role in HTLV-I infection as a modulator of growth factors, cytokines, their receptors, and proto-oncogenes.NF-κB1 results in lower protein levels of the NF-κB p50 subunit. NF-κB1 94ATTG ins/del polymorphism was associated with risk of cancerNF-κB1 94ATTG ins/del polymorphism and HTLV-I for the risk of total death or incidence of cancer and atherosclerosis-related diseases.NF-NF-κB1 94ATTG ins/del polymorphism was not in the HWE. The HWE deviation has two possible explanations: genotyping error or population stratification. Since its MAF (0.364) was not different between Japanese subjects overall and Japanese subjects from the Amami region (0.357),Several limitations of this study should be considered. First, the intermediate markers of HTLV-I associated diseases, such as anti-HTLV-I antibody titers, provirus load, or anti-Tax protein antibody, were not included. However, the present study used inflammatory gene polymorphisms to identify susceptible HTLV-I carriers, while a previous study reported a significant association between provirus load and inflammatory gene polymorphism.TNF-α 1031T/C polymorphism and HTLV-I for the risk of atherosclerosis-related disease incidence, but not for the risk of total death and cancer incidence. Further follow-up study is warranted to understand the protective factors against developing diseases among susceptible HTLV-I carriers.In conclusion, the present study found an interaction between the None declared.


Scientific Reports7: Article number: 42023; 10.1038/srep42023 published online: 02102017; updated: 03222017.This Article contains typographical errors.Boolean networks’, “In Boolean networks, variables can only take one of two possible values, 0 or 1, and their dynamics is described byIn the methods section under subheading ‘xi(t + 1) represents the value of variable i at the time t + 1 as a Boolean function Fi of its n regulators xi(t), …, xn(t) at the current time”.where should read:“In Boolean networks, variables can only take one of two possible values, 0 or 1, and their dynamics is described byxi(t + 1) represents the value of variable i at the time t + 1 as a Boolean function fi of its n regulators xi(t), …, xn(t) at the current time”.where Non-hierarchical and non-unidirectional interactions greatly increases the dynamical diversity of pathways’,In the Results section under subheading ‘inputs is the set of inputs”.“Because the inputs follow the identity function, the minimum number of attractors is equal to 2|inputsl|, where should read:inputs|| where inputs is the set of inputs”.“Because the inputs follow the identity function, the minimum number of attractors is equal to 2


We characterised and reported the first full-length genomes of Human T-cellLymphotropic Virus Type 1 subgroup HTLV-1aD (CV21 and CV79). This subgroup is one ofthe major determinants of HTLV-1 infections in North and West Africa, and recombinantstrains involving this subgroup have been recently demonstrated. The CV21 and CV79strains from Cape Verde/Africa were characterised as pure HTLV-1aD genomes,comparative analyses including HTLV-1 subtypes and subgroups revealed HTLV-1aDsignatures in the envelope, pol, and pX regions. These genomes provide originalinformation that will contribute to further studies on HTLV-1a epidemiology andevolution. Deltaretrovirus genus and is the first oncogenic human retrovirusdiscovered (The Human T-cell Lymphotropic Virus Type 1 (HTLV-1) belongs to the Retroviridae family inthe scovered . HTLV-1 scovered . A recenscovered . ItsgenMost genetic and evolutionary studies on HTLV-1 were based on partial genome sequences,particularly the LTR region. However, complete genome sequences of the most prevalentHTLV-1a subgroups (except HTLV-1aD) are currently available .The phylogenetic analysis, based on the LTR region, revealed the presence of the HTLV-1aDsubgroup in Cape Verde Archipelago in Africa .In this study, we present the complete genome sequence of two HTLV-1aD strains from CapeVerde along with genome-wide comparative analyses including HTLV-1a prevalentsubgroups.Supplementary Table I). Amplicons were sequenced in theSanger Platform at Fiocruz using the BigDye Terminator v3.1 Cycle sequencing kit . Genome sequences were deposited in the GenBank under theaccession numbers: KX430030and KX430031.The genome of the CV21 and CV79 strains from Cape Verde were amplified through nestedpolymerase chain reaction (PCR) using degenerate HTLV primers and the PCR-basedgenome-walking strategy  andsubtypes (b and c)  showed tSupplementary TablesII-III).HTLV-1aD is characterised by unique amino acids residues, when compared with othersubgroups and subtypes which share the same conserved residue in the following positions:gp46 env A59V; gp21 env L454F; TR K486R; p30 S105N; and HBZ isoforms M/I and V/A.Regardless of presenting unique residues, the charges and polarity properties of theseresidues were maintained in HTLV-1aD. In some other positions, HTLV-1aD present specificamino acid (aa) residues, which is contrasting to what is observed in other subgroups andsubtypes that do not share the same conserved residues in the following positions: TRV/A310I; p12 D/N/E/A26K; HBZ SP2 isoform L/P13R. Moreover, HTLV-1aD specific substitutionsin p12 and HBZ showed different charge and polarity aa properties(The two HTLV-1aD complete genomes presented in this study represent original informationabout a subgroup that is prevalent in African countries. These genomes provide informationthat will contribute to further studies on HTLV-1a epidemiology and evolution.


PTK7 (protein tyrosine kinase 7) is an evolutionarily conserved transmembrane receptor regulating various processes in embryonic development and tissue homeostasis. On a cellular level PTK7 affects the establishment of cell polarity, the regulation of cell movement and migration as well as cell invasion. The PTK7 receptor has been shown to interact with ligands, co-receptors, and intracellular transducers of Wnt signaling pathways, pointing to a function in the fine-tuning of the Wnt signaling network. Here we will review recent findings implicating PTK7 at the crossroads of Wnt signaling pathways in development and disease. PTK7 (protein tyrosine kinase 7) is an evolutionarily conserved transmembrane receptor with a broad range of functions in tissue development and homeostasis. Originally identified as a gene upregulated in colon carcinoma cells and accordingly named colon carcinoma kinase 4 (CCK-4)  and Disheveled (Dsh) were identified. Other core PCP proteins include the four-pass transmembrane protein Van Gogh , the atypical cadherin Flamingo  and intracellular components like Prickle (Pk) and Diego (Dgo). These proteins confer intra- and intercellular signaling, thereby aligning PCP in neighboring cells. Complementary studies in vertebrates revealed that these core PCP proteins are also required for the polarization of vertebrate tissues, like the orientation of hair follicles in the epidermis or the sensory hair cells in the inner ear , and off-track2 (otk2). These two genes, which are the result of a tandem gene duplication, function redundantly in the tubular morphogenesis of the male ejaculatory duct, leading to male sterility in the otk, otk2 double mutant  mutant zebrafish exhibit vertebral abnormalities at larval stages, phenotypically resembling congenital scoliosis. Further, zygotic ptk7 (Zptk7) mutants show late onset spinal curvatures consistent with the idiopathic form of scoliosis  is apparent at birth and involves abnormal vertebrae development, while idiopathic scoliosis is diagnosed during adolescence and does not show vertebral malformations. During the last two decades our understanding of the function of PTK7 has significantly advanced. Diverse biological processes that are regulated by PTK7 have been identified and its role in the establishment of polarity and coordinated cell movements has been acknowledged. Recent publications shed light on a literary “complex” function of PTK7 in Wnt signaling. While its role in non-canonical PCP signaling has been confirmed in different animal model systems and biological contexts, its function with respect to canonical Wnt signaling remains controversial. Possibly, these contradictory findings can be explained by the cell type-specific formation and subcellular localization of distinct co-receptor complexes. Further characterization of the formation and dynamics of these ligand-receptor complexes may help us to understand how PTK7 affects development as well as disease-related processes.Drosophila and the cancer section and edited the manuscript; HB wrote the section on scoliosis and designed the two tables.AB conceptualized and wrote the main part of the paper. AW wrote the 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.


Retroviruses of human T-lymphotropic viruses (HTLV-1 and HTLV-2) have been demonstrated to be endemic in the north-eastern region of Iran. This study was aimed to determine the HTLV-1 and HTLV-2 prevalence among healthy individuals in Neyshabur City during 2010-2014.A total of 8054 blood samples were collected from healthy participants in Neyshabur, North-Eastern Iran. The blood samples were screened for the presence of specific antibodies against HTLV-1 and HTLV-2 by using ELISA according to the manufacturer’s instructions.The overall seropositivity rate for HTLV-1 and HTLV-2 was found to be 6.55% (528 out of 8054) among participants.Both HTLV-1 and HTLV-2 were demonstrated to be at a high rate in healthy individuals. However, a smaller number of asymptomatic carriers were found in this study, as compared to those identified in previous investigations in the city. Human T-lymphotropic viruses (HTLV-1 and HTLV-2), classified in the retroviridae family, are among the first identified species. HTLV-1 A total of 8054 healthy individuals from Neyshabur, North-Eastern Iran, were included in this study. Serum samples (5 ml) were prepared from the individuals and stored at -20°C until the ELISA test.Serum samples were screened for the presence of specific antibodies against HTLV-1 and HTLV-2 by ELISA  according to the manufacturer’s instructions.t-test. A P<0.05 was considered to be statistically significant.The SPSS software (version 20) was employed to analyze all data using chi-square and Of 8054 healthy individuals participated in the study, 1565 (19.4%) and 6489 (80.6%) were males and females, respectively. As shown in et al.[et al.[Previous studies have revealed that HTLV-1 is endemic in North-Eastern Iran. Anotheret al.. It seeml.[et al. in MashhIn the current study, over 3% of healthy individuals were positive for HTLV-1 in all five years. To the best of our knowledge, there is a small number of published data regarding HTLV-2 prevalence in Iran. Also, a lower rate of positive HTLV-1 infection was identified in the present investigation, when compared to a previously study in Neyshabur. This fiIn the present study, a high rate of HTLV-1 among serum samples was detected using the ELISA test among healthy individuals in Neyshabur city during 2010-2014. The results from this study emphasize that HTLV is still an important endemic disease in Neyshabur. More importantly, the prevalence of HTLV-1 in Neyshabur was detected to be higher than other city (Mashhad) in all duration of this study, though being in a decreasing status compared to the previous reports.


The growth hormone receptor (GHR), although most well known for regulating growth, has many other important biological functions including regulating metabolism and controlling physiological processes related to the hepatobiliary, cardiovascular, renal, gastrointestinal, and reproductive systems. In addition, growth hormone signaling is an important regulator of aging and plays a significant role in cancer development. Growth hormone activates the Janus kinase (JAK)–signal transducer and activator of transcription (STAT) signaling pathway, and recent studies have provided a new understanding of the mechanism of JAK2 activation by growth hormone binding to its receptor. JAK2 activation is required for growth hormone-mediated activation of STAT1, STAT3, and STAT5, and the negative regulation of JAK–STAT signaling comprises an important step in the control of this signaling pathway. The GHR also activates the Src family kinase signaling pathway independent of JAK2. This review covers the molecular mechanisms of GHR activation and signal transduction as well as the physiological consequences of growth hormone signaling. C-mannosylation. For the IL-21R, the WSXWS has been shown to be mannosylated at the first tryptophan where the sugar chain appears to form structurally important interactions that bridge the two fibronectin domains  and PI 3-kinase/Akt are also activated by GHR (The growth hormone receptor (GHR) is a member of the class I cytokine receptor family, which includes more than 30 receptors such as the prolactin receptor (PRLR), erythropoietin receptor (EPOR), thrombopoietin receptor (TPOR), granulocyte-macrophage colony-stimulating factor receptor, interleukin-3 receptor, interleukin-6 receptor, and interleukin-7 receptor , 2. GHR  domains , 5. GHR  domains . GHR alsd by GHR . The convia its “site 2” interactions  . The N-tvia autophosphorylation of two critical residues in the JH2–SH2 linker region, Ser523 and Tyr570, which play an important role in regulating the activity of the KD  followed by a kinase domain (KD), also known as JH2 and JH1, respectively . The KD f the KD , 83.trans  and diacylglycerol which leads to an increase in the cytoplasmic calcium ion concentration-activating Ras guanine nucleotide exchange factor RasGRP1. This process results in RAS activation and initiation of the ERK1/2 signaling pathway . T. T109).  pathway , 112. Mu pathway . Studies pathway –117. In  pathway –120. The pathway , 122.Negative regulation of cytokine receptor signaling is critical to the correct function of cytokine receptors as dysregulated negative regulation of signaling can lead to disease states. Negative regulators of JAK–STAT signaling can be grouped into three main families, and this includes the suppressor of cytokine signaling (SOCS) protein family, protein tyrosine phosphatase (PTP), and protein inhibitor of activated STAT (PIAS). Apart from these protein families, the SH2-domain containing LNK (SH2B3) adaptor protein has been shown to be an important negative regulator in the JAK–STAT pathway. These negative regulators act in different ways to suppress JAK–STAT signaling. They can target JAKs, STATs, and/or receptors and may also involve other proteins of the proteasomal degradation pathway . The varThe SOCS protein family consists of SOCS1–7 and CIS. SOCS genes are translated as a result of cytokine-induced activation as part of the JAK–STAT negative feedback loop. SOCS proteins bind JAKs or their receptors and act by (i) abolishing the kinase ability of JAK, (ii) abolishing the ability of JAK to bind to the receptor, (iii) blocking STATs from being recruited to the receptor, and (iv) mediating the ubiquitination of JAKs and STATs for degradation .−/−socs2 mice are 30–40% larger than WT littermates while other SOCS knockouts (KOs) do not significantly affect size : (i) non-receptor PTPs such as SHP1 and SHP2, (ii) receptor tyrosine phosphatases such as CD45, (iii) dual-specificity phosphatases, and (iv) low molecular weight phosphatases . AccordiPTPN6; also known as HCP, PTP1C, SH-PTP1, and Hcph) is generally expressed in hematopoietic cell lineages whereas SHP2  has ubiquitous expression. Both SHP1 and SHP2 have high sequence and domain homology whereby they contain two SH2 domains at their N-terminal region, denoted as N- and C-SH2 domains, and a PTP catalytic domain. They also possess a C-terminal tail with tyrosine residues that can be phosphorylated to regulate PTP activity  inhibiting the DNA-binding activity of STATs, (ii) substrate SUMOylation, or (iii) recruit other transcriptional co-repressors. While PIASs are able to associate with STATs, they are also able to regulate other transcription factors including SMADs, androgen receptor, c-JUN, and p53. The PIAS family of transcriptional co-regulators consist of PIAS1, PIASxα, PIASxβ, PIAS3, PIAS3L (PIAS3β), PIASy, and PIASyE6. Although PIAS proteins are known as negative regulators of JAK–STAT signaling, their specific role in GH-mediated signaling is not well studied.SH2B3 gene and is part of the family of SH2-containing adaptor protein that includes SH2B (or SH2B1) and APS (or SH2B2). However, these adaptor proteins have different functions as the binding of SH2B or APS to JAK2 enhances its kinase activity whereas LNK binding to JAK2 inhibits its kinase activity . T. T154). The anti-insulin action of GH is very well established  as liverGH receptors and IGF-1 receptors are abundant on not only adipocytes but also preadipocytes, fibroblasts, various immune cells, and endothelial cells –188. TheThe significance of the GH/IGF-1 axis in the muscular system was highlighted when it was found that calcineurin (CaN), a calcium-calmodulin-dependent protein phosphatase that activates slow isoform of troponin I and myoglobin genes, was regulated by IGF-1 and that inhibiting CaN blocked IGF-mediated hypertrophy of cultured skeletal muscle cells . In addiigf1 gene in the liver and a consequent decrease in systemic IGF-1 did not affect the animal’s body and femoral growth  has been shown to correlate with high IGFBP-1 and low IGFBP-3, with IGFBP-3 being a major limiting factor for the reported low circulating IGF-1 levels . A similCirculating GH/IGF-1 levels peak in the second decade of life , 243 witAs GH/IGF signaling induces growth mechanisms, inhibiting this signaling should suppress cell proliferation and a reduction in the cancer risk; however, other important mechanisms that may be affected by this process include a decrease in immune responses and reduced favorable metabolic adjustments . MoreoveGHR that correlated with an increased risk of lung cancer  followed by breast and colorectal cancers  have obstructed growth with a similar phenotype observed in mice with ghr KO (ghr (at amino acid position 391), which eliminates all GH-mediated STAT5 signaling while still allowing for JAK2, STAT1, and STAT3 activation (Postnatal growth of mice has been shown to rely on signaling mediated by JAK2 and STAT5, downstream from the GHR , 256. Mih ghr KO , 110. A Although we have learnt a vast amount on the molecular mechanism of GHR activation, signaling, physiological aspects, and roles of GH signaling in disease states, there is still much to learn. In particular, this is due to the wide range of physiological roles that GH has, making it an important player in many biological conditions and diseases. At the molecular and cellular level, the activation and role of SFK signaling is still currently not well understood. As our knowledge of the molecular signaling nature of GHR increases, our ability to specifically target the receptor and its signaling pathways for a diverse range of therapeutic purposes should also increase.All the authors contributed equally. All the authors contributed to drafting and editing the manuscript. All the authors contributed to figures and tables.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.