Wnt signaling pathways constitute evolutionarily conserved signaling pathways that control a wide range of cellular processes including proliferation, migration, differentiation and morphogenesis during development, maintenance of adult tissue homeostasis, and organ regeneration. Thus, it is not surprising that dysregulation of the Wnt pathway has been associated with many human diseases including cancer, congenital malformations, degenerative diseases, and metabolic disorders. At this point, detailed understanding of underlying Wnt-mediated cellular responses is essential for the development of effective therapies against Wnt-related diseases. The plasma membrane (PM) plays a key role in the regulation of cell signaling and thus has been frequently investigated for its molecules as drug targets. Owing to its essential roles in activation of Wnt-receptor complex and initiation of Wnt signaling, the PM has been investigated thoroughly for the regulatory role of its composition and lipid order in pathway regulation. A prominent feature of unhealthy cells such as cancer cells is that they significantly differ from healthy cells with respect to their PM composition and lipid organization. Since the complex interactions of Wnt proteins and their receptors are tightly dependent on the composition and organization of the PM, a better understanding of the molecular mechanisms underlying Wnt-receptor complex formation is essential for identification and testing of novel attractive drug targets for Wnt-related human diseases including cancer. In this Research Topic (RT) with 15 papers, we aimed to collect the review and research articles that provide an overview of the recent developments that enlighten the molecular interaction mechanisms of the Wnt-receptor complex components and pathway modulators as well as how misregulation of these interactions associate with signaling activation and human diseases. We have also included recent scientific progress that we believe that understanding these mechanisms will greatly contribute to the development of novel therapies that target Wnt pathways at the PM.Suthon et al. have provided a comprehensive view of the mechanistic roles of the non-canonical ligand WNT5B on general development, bone physiology, adipogenesis, pancreas development, heart development, central nervous system . They also review the potential use of therapeutic approaches that functionally antagonize Wnt signaling at the level of ligand secretion or receptor binding. Dhasmana et al., have presented the results of their elegant work on characterization of the role of lipid modifications of the canonical Wnt ligand Wnt3 in membrane organization, secretion, interaction, and signaling activity by exploiting the zebrafish model. By substituting the homologous cysteine and serine residues of zebrafish Wnt3 with alanine, they have revealed that Wnt3 is lipidated at both residues. Lipidation at either of the residues is sufficient for its secretion and proper interaction with the membrane, while the lipid modification at serine is indispensable for receptor interaction and signaling.For this RT, we have received several papers that present detailed analyses of the structure and functional roles of various Wnt ligands, receptors and co-receptors in development and disease. In their review, Pascual-Vargas and Salinas have elaborately reviewed the molecular function of Frizzled receptors in the mammalian CNS with a special focus on its roles in axon guidance, dendritic morphogenesis, synapse formation and postnatal hippocampus development. The authors have also discussed in detail the post-translational modifications, namely N-linked glycosylation, phosphorylation, and ubiquitination/deubiquitination, of Frizzled receptors regarding their modulation of the receptor function in neuronal connectivity through control of its localization and stability at the PM. Another review by Ren et al. has systematically examined the similarities and divergences between the canonical Wnt co-receptors LRP5 and LRP6 with respect to their expression patterns in human tissues and organ development. They highlight that, in contrast to the high homology between LRP5 and LRP6, they are expressed differentially in diverse tissues and organs in embryonic development and adulthood and that LRP6 is functionally more potent and unique via coupling both Wnt with Hippo signaling pathways. The authors have also discussed how Wnt signaling is activated after kidney injury and in renal tissues in diabetes. In their brief research report, Haack et al. have analyzed whether and how the quantitative ratio between membrane receptors and membrane microdomains (rafts) influence LRP6 phosphorylation and WNT/β-catenin pathway activation. By using a computational modeling approach and considering microdomains as single compartments in the membrane, they have found that LRP6 phosphorylation and downstream Wnt signaling activity decreases when the number of raft compartments exceeds the number of pathway specific membrane proteins. This reveals that pathway specific targeting and sorting are critical to restrict the receptor/raft ratio and signaling activation. Jeong and Jho have reviewed activation of LRP6 and its influence on β-catenin-dependent and -independent signaling pathways. They have also discussed in detail how LRP6 dysregulation is associated with diseases including cancer, neurodegeneration, metabolic syndrome, inflammation, and skeletal disease. Shi has thoroughly examined the functional roles of Disheveled in Wnt signaling pathways in gastrulation and neurulation processes by focusing on the vertebrates mouse, Xenopus, and zebrafish. The author specifically emphasizes the influence of Disheveled on maternal contribution for axis patterning, cell fate specification and morphogenetic movements.Azbazdar et al. summarizes how membrane composition and lipid order tightly controls formation of the Wnt-receptor complex, its interactions with the pathway modulators and activation of Wnt signaling pathways. Moreover, the study gives a detailed overview of the involvement of the ligands, receptors, coreceptors and extracellular or membrane-bound modulators of Wnt pathways in progression of lung, colorectal, liver, and breast cancers that have been associated with abnormal activation of Wnt signaling.A comprehensive review by Yong et al. have unraveled the stimulatory role of adiponectin, a secreted protein produced by adipocytes, on the expression of adiponectin receptors as well as β-catenin and MAPK signaling pathways in cementoblasts in the presence of compressive forces. They have also demonstrated that β-catenin is blocked by MAPK inhibition and rescued by adiponectin.In their original research, Azar and Lim have summarized the individual roles of the Wnt pathway components, namely Wnt ligands, β-catenin, destruction complex component APC and transcription factor TCF7L2 in development of metabolic tissues. Moreover, they have discussed the contributions of these components to the whole-body metabolism and potential roles in development of metabolic diseases such as diabetes and obesity.In addition to its well-known roles in development, growth and tissue patterning, Wnt signaling in mammals is necessary for regulation of energy homeostasis. In their elegant review, Hachim et al. have found that canonical Wnt signaling is activated and required for the asthmatic bronchial epithelial cells to respond to this stimulus. These cells have activated senescence mechanisms in a Wnt-dependent manner. Interestingly, fibroblasts from asthmatic patients were not responsive to inhibition of Wnt signaling.In their study to understand the role of proliferative stress, Karabicici et al. have shown that the multi-kinase inhibitor regorafenib activates Wnt/β-catenin signaling selectively in hepatoblast-like HCC cell lines. Strikingly, Wnt/β-catenin signaling protects these cells from regorafenib-induced apoptosis. In contrast, long-term regorafenib-treated resistant cells have reduced Wnt/β-catenin activity and enhanced TGF-β activity with a concomitant increase in expression of cancer stem cell markers. Importantly, regorafenib treatment and TGFβ-R1 inhibition have collectively decreased colony formation ability and promoted cell death in resistant spheroids. These results demonstrate that HCC tumors with aberrantly activated Wnt/β-catenin pathway are more resistant to regorafenib, suggesting that regorafenib administration in combination with pathway inhibition can enhance the drug-induced cell death. On the other hand, acquired regorafenib resistance in HCC can be overcome with the combined use of TGF-β pathway inhibitors together with regorafenib.Finally, To conclude, this RT provided us with the opportunity to collect recent data and perspectives from researchers whose studies on Wnt signaling pathways span over a wide range of disciplines. By enabling us to review the recent findings and advances in activation and regulation of Wnt signaling in health and disease, we believe that this RT will greatly contribute to our understanding of pathway mechanisms and the development of new therapeutic approaches for Wnt-related human diseases.GO has written the Editorial manuscript.GO Lab has been supported by the Scientific and Technological Research Council of Turkey  and EMBO Installation Grant (IG 3024).The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


There were no significant differences in ankle kinematics between the groups before the intervention. However, after the intervention, healthy controls demonstrated significantly less ankle inversion between 0–9% (p = 0.009) and 82–87% (p = 0.011) of the stance phase. Furthermore, a significant within-group difference was observed only in the control group, demonstrating increased ankle eversion between 0–7% (p = 0.011) and 67–81% (p = 0.006) of the stance phase after the intervention. Peroneal EMG activity did not differ between groups or measurements. These findings, which demonstrate that peroneal FES can induce ankle kinematics adaptations during gait, can help to develop future interventions for people with CAI.Chronic ankle instability (CAI) is a common condition that may develop after an ankle sprain. Compared with healthy individuals, those with CAI demonstrate excessive ankle inversion and increased peroneal electromyography (EMG) activity throughout the stance phase of gait, which may put them at greater risk for re-injury. Functional electrical stimulation (FES) of targeted muscles may provide benefits as a treatment modality to stimulate immediate adaptation of the neuromuscular system. The present study investigated the effect of a single, 10 min peroneal FES session on ankle kinematics and peroneal EMG activity in individuals with ( Chronic ankle instability (CAI) is a condition characterized by repetitive episodes of the perception that the ankle is unable to support continued locomotion. This may be accompanied by pain, local muscle weakness, and/or reduced range of motion in the ankle joint. CAI is diagnosed when recurrent ankle sprains persist for more than 1 year after the initial injury ,3. CAI hWhen compared with healthy subjects, individuals with CAI display sensorimotor control deficits such as reduced postural stability ,21, reduNeuromuscular electrical stimulation (NMES) is a potential therapeutic agent to overcome AMI . When apFES provides coordinated, rhythmic stimulation, in a pattern that imitates normal movement and therefore contributes to the recovery of muscle control . FurtherIt has been previously shown that FES provided in concurrence with gait training is more beneficial in restoring muscles strength and control after anterior cruciate ligament reconstruction when compared with NMES . FurtherThe objective of this study was to investigate the immediate carryover effects of a single gait training with peroneal FES on ankle kinematics and peroneal activity in individuals with and without CAI. Completing these objectives should convey primary information for future interventional studies that will explore the potential benefits of FES in CAI rehabilitation. Furthermore, comparing FES effects between these groups may expand the understanding of the impaired gait in CAI. We hypothesized that applying peroneal FES during 0–80% of the stance phase of gait will decrease ankle inversion angle after heel contact and decrease muscle activation in both groups .The sample size for this study was determined based on a power analysis simulation, using the Power1D package for Python . We usedExclusion criteria for all groups were a history of lower extremity fracture that required realignment, surgical procedures, or other pathological conditions in the lower extremities, and vestibular or neurological disorders. Participants were recruited by advertising the study in the university facilities and social media. All participants provided written informed consent before participating in the study. Ariel University Institutional Review Board approved the study . The study was pre-registered at ClinicalTrials.gov (Identifier: NCT04314960).The study was conducted during a single session at the Neuromuscular and Human Performance Laboratory, at Ariel University. Demographic variables  were collected from all participants. Additionally, CAIT scores were collected for each limb separately, as well as the Foot and Ankle Ability Measure (FAAM) . CAIT is2 Challenger, Taipei, Taiwan) for 30 s before and immediately after a 10 min peroneal FES. To synchronize kinematic and muscle activation data collection, an external trigger was used to start both recordings simultaneously. Participants walked barefoot and wore tight, black sports pants and t-shirts. To determine gait speed, the treadmill was set at 3 km/h (0.83 m/s). Then, participants were instructed to increase the speed (using a remote control and without seeing the treadmill screen) until reaching a comfortable, self-selected pace (SSP). The instruction given was “walk at the speed you would walk in the street if you were slightly in a hurry”. After determining SSP, walking speed was increased by 20% (SSP + 20%) for gait measurement, since it was previously reported that people with CAI demonstrate greater ankle inversion as speed increases [Ankle kinematics and peroneal activity were assessed during walking on a motorized treadmill  sampled at 100 Hz was used to obtain three-dimensional ankle kinematics. Data were exported to Visual 3D software , and processed through a 6 degree-of-freedom anthropometric model. Ankle angles in the frontal plane during walking were calculated using the Cardan rotation sequence . The rawPeroneal electromyography (EMG) activity was collected using a wireless EMG system  at 2000 Hz. Before data collection, participants’ skin was shaved and cleaned with alcohol to minimize impedance. Electrode placement was performed according to Surface Electromyography for the Noninvasive Assessment of Muscles (SENIAM) guidelines . Before Peroneal stimulation was delivered using an FES system . The system consists of three components that communicate via radio frequency signals: (1) a stimulator, (2) a gait sensor that is placed under the heel, and (3) and control unit. An algorithm analyzes the gait sensor’s data to detect gait events  in real time . The par2Χ tests were used for continuous and categorical variables, respectively. Time series data  were plotted throughout the stance phase with their corresponding 95% confidence intervals (CI) [t-tests with Bonferroni correction for multiple comparisons were conducted, to assess the within-group (pre- vs. post-intervention) and between-group  differences. The significance level was set as p < 0.05. Python 3.8 was used for all data processing. Statistical analysis was conducted using R v.4.0.3  and SPM1d v.0.4 package for Python 3.8 [To compare between-group baseline characteristics, nonparametric Mann–Whitney U tests and als (CI) . A signials (CI) ,63. Addithon 3.8 .p < 0.001). The average FAAM–ADL and FAAM–sports scores were 90.4 ± 10.4 and 62.7 ± 15.0 in the CAI group and 99.4 ± 1.6 and 98.1 ± 3.5 in the control group (p < 0.001)In total, 24 participants with CAI  and 24 controls  completed this study. Their demographic characteristics are presented in p = 0.024) and between 77% and 91% (p = 0.018) of stance, and significant group x time interaction between 39% and 82% (p < 0.001) of stance. No significant main effect was found for time. Post hoc between-group and within-group comparisons are displayed in p = 0.009) and between 82% and 87% (p = 0.011) of the stance phase. No within-group differences in ankle inversion angle were demonstrated in the CAI group. However, after the intervention, the control group demonstrated more ankle eversion than before the intervention between 0% and 7% (p = 0.011) and between 67% and 81% (p = 0.006) of the stance phase  = 16.64, p < 0.001) but not for group or group x intervention interaction. For RMS, a significant main effect was found for group  = 0.68, p = 0.41), as well as significant group × intervention interaction effect  = 4.3, p = 0.04). A trend for intervention effect was also found  = 3.04, p = 0.08).After reviewing the study results, a secondary post hoc analysis was conducted to explore a possible mechanism for the between-group difference in the kinematic adaptation to FES. We assumed that this finding might be the result of an earlier onset of peroneal fatigue in the CAI group (see the Discussion Section below). Muscle fatigue can be assessed with further signal processing of EMG data. A reduction in EMG median-frequency (MDF) and an increase in the signal root mean square (RMS) have been suggested to indicate muscle fatigue . TherefoThe current research investigated the effect of peroneal FES on gait patterns of individuals with CAI. Specifically, we examined the immediate carryover effects of peroneal FES on ankle frontal plane kinematics and peroneal activity throughout the stance phase of gait. Contrary to our hypothesis, peroneal FES did not normalize the walking pattern of people with CAI. However, healthy controls demonstrated increased ankle eversion angle at early and late stance after the intervention. Peroneal EMG activity was not affected by the intervention in either group. To the best of our knowledge, this study is the first to report these results.Our results show that immediate kinematic adaptation was observed in the control group in the current study but not in the CAI group. The different adaptation to FES of subjects with and without CAI may be related to muscle fatigue resulting in a decrease in power production in response to repeated contractile activity. Compared with voluntary muscle contraction, ES may induce greater muscle fatigue due to altered muscle recruitment patterns . During The assumption that muscle fatigue influenced the different adaptation patterns between the groups was investigated in a secondary post hoc analysis. Assessing muscle fatigue using EMG signal can be performed by inspecting the time-domain features  or the frequency-domain features  . Fast-tw2) electrodes to apply the stimulation. While larger electrodes may produce less discomfort, they may recruit a larger portion of motor units, resulting in earlier fatigue onset. Future studies should address these issues, by applying shorter FES durations and decreasing electrodes’ size.ES-induced muscle fatigue can be reduced by manipulating several factors, such as stimulation parameters, training duration, and electrode position and size . The FESAn additional factor that might have influenced the results of the present study is related to the stimulation sensation. The sensation of electrical current during FES may contribute to sensorimotor excitability and enhanced muscle contraction . Burcal Individuals with CAI demonstrate a more inverted ankle position at early stance, which may put them at greater risk for re-injury ,56. An iSeveral studies have investigated EMG patterns in individuals with CAI and reported increased peroneal muscle activation during the stance phase of gait, compared with healthy controls ,29,30. SThe findings of this study should be interpreted with some limitations. Our control group included five subjects who had a history of an ankle sprain. It may be argued that these individuals could be referred to as ankle sprain copers, not healthy controls. However, we do not believe that the inclusion of these subjects had a major effect on the results, as all ankle sprains occurred long before participating in the study, and all subjects reported no activity limitation due to their ankle history. The question of how long an individual should be considered as an ankle sprain coper after an injury is of interest for CAI studies and should be discussed in future studies. Furthermore, in this study, we only examined the immediate carryover effects of FES. This type of research is not suitable to draw clinical conclusions, but it may suggest a different adaptational pattern to FES in individuals with CAI. Several confounding variables may have influenced our results. For example, direct measurement of peroneal muscle strength and endurance could have assisted in accepting or refuting the presumption that muscle fatigue is the differing factor between the groups. Ankle characteristics such as passive instability tests and passive range of motion measurements could also better describe the study’s sample. Future studies should try to control for these factors by directly measuring fatigue and neural excitability before and after the FES intervention. Different intervention protocols should also be used to better understand the different adaptation patterns between individuals with and without CAI. Studies that will evaluate the therapeutic effect of several training sessions are also warranted.To the best of our knowledge, this is the first study that investigated the immediate carryover ankle kinematics effects of FES in individuals with CAI. The findings of this study, which demonstrate ankle kinematics modifications in healthy controls, should encourage future research to understand how to successfully transfer these findings to people with CAI.


The princeps pollicis artery (PPA) is typically a direct branch off the deep palmar arterial arch. Identified is a 90-year-old female cadaver in which the right hand has a duplicated PPA and radialis indicis (RI) artery. These vessels originate from the superficial palmar arterial arch as variant vessels as well as from the deep palmar arterial arch. The superficial arch appears in its classic pattern, while the duplicate PPA and RI present at the radial aspect of the superficial arch in the volar first web space with clear communication to the superficial radial artery. There are many common surgical procedures that require precise knowledge of the first web space, such as Dupuytren's contracture release, trigger thumb release, and syndactyly release at the first web space. Further, percutaneous pinning of fractures at the base of the thumb may pose an inherent risk to the underlying vessels. Understanding these variations of hand vasculature is of clinical significance in disciplines such as orthopedic surgery, plastic surgery and vascular surgery. This case report has been presented as a poster at the 2020 Student Osteopathic Surgical Association Spring Convention in Des Moines Iowa on February 20, 2020, and the Consortium for Excellence in Medical Education in April 2020, online. Blood supply to the hand is a result of two anastomosing arches, which arise from the two main arteries of the forearm - the ulnar artery and radial artery. The ulnar artery produces the superficial palmar arterial arch, which is the primary branch responsible for supplying the palm and fingers. The deep palmar arterial arch is a terminal branch of the radial artery that is responsible for supplying the deep hand. The princeps pollicis artery (PPA) and radialis indicis (RI) are typically direct branches off the deep palmar arterial arch. The PPA has been used in reference to the first palmar metacarpal artery or simply the primary source of blood supply to the thumb .The PPA is typically one of the terminal branches of the radial artery. It continues to travel distally along the ulnar surface of the first metacarpal and dorsal to the origin of the adductor pollicis muscle. As the artery traverses, it sends off branches that feed the proximal half of the first metacarpal. Upon reaching the neck of the first metacarpal, it then sends off two circumflex branches - the ulnar circumflex branch and radial circumflex branch. The ulnar circumflex branch runs inferior to the tendon of the adductor pollicis to anastomose with a branch of the dorsal metacarpal artery. The radial circumflex branch runs laterally between the insertions of the opponens pollicis and the flexor pollicis brevis muscles to anastomose with a branch of the dorsal arteries of the back of the metacarpal bones . The PPAIt is not uncommon to have anatomical variations with blood supply to the hand, which has been noted in multiple studies -3. AccorIdentified is a 90-year-old female, with no obvious deformity of the hands or bilateral upper extremities and no obvious surgical procedures performed on the hands or bilateral upper extremities. The right hand is shown to have a duplicate PPA as well as a duplicate RI emerging from the superficial palmar arch in communication with the superficial radial artery Figure . This caAn initial approach to the vasculature of the right hand was done volarly. After superficial dissection of the palmar fascia, the superficial palmar arch was visualized. In addition, the superficial radial artery was visualized at the radial aspect of the wrist directly communicating with the superficial arch. A small arterial branch, approximately 2 mm in length was visualized emerging from the superficial palmar arch in the volar first web space further bifurcating into a PPA and RI. This PPA continued along the ulnar aspect of the first metacarpal Figure . The RI In an attempt to ascertain any contribution of blood supply to the thumb from the deep palmar arch, the deep branch of the radial artery was dissected at the anatomical snuffbox. The artery was followed distally into the dorsal first web space. At the level of the metacarpotrapezial joint, the deep radial artery gave off three branches: (1) the PPA, which courses along the ulnar side of the first metacarpal extending distally towards the phalanges of the thumb. 2) The deep palmar arch traveling in an ulnar direction across the volar hand at the level of the metacarpals. (3) The RI coursing along the radial aspect of the second metacarpal  . If a clTreatment such as thenar flap surgery and thumb amputations involving replantation may be affected by vascular anomalies of the hand as found here. In the case of a duplicate PPA, one must consider more blood flow to a thenar flap through the duplicate artery. The proposed time to keep a thenar flap intact is two weeks . This tiIn the incidence of total thumb amputation, great toe transfers are the treatment of choice for plastic surgery; however, these transfers have been reviewed as complex . One comCadaveric dissection has been a common practice by medical students for centuries. In contemporary times, medical student preparation prior to dissection may include reading dissection manuals, watching online dissection videos, and using medical atlases. Typically, dissection laboratory manuals and basic human anatomy atlases do not show branches from the superficial palmar arch to the thumb. The volar side of the hand has many superficial structures that must be carefully dissected, ergo without knowledge of this PPA variation it could potentially be dissected out in the process, therefore, potentially contributing to why this anatomical variation may be under reported in literature. More research should be done with special attention to the superficial radial artery to see the incidence of this specific variation. As technology advances, more medical schools are moving away from traditional cadaver labs and towards virtual platform. This may reduce students’ tactile skill as well as their observation of anatomy in a “non-textbook” manner.This case report explores an incidence of duplicated PPA and RI occurring from the superficial palmar arch. An intact PPA and RI artery are described as anatomically expected from the deep palmar arch. To the best of the authors’ knowledge, this anatomic anomaly has not been fully described in medical literature except for in one other case report, which lacks description of the full vasculature to the thumb. Much of the discussion surrounds the clinical implications of such anomaly. Further research on cadavers with special attention to the volar first web space may help establish an increase prevalence of vascular variations of the superficial and deep palmar arches.


Cancer can be considered as a communication disease between and within cells; nevertheless, there is no effective therapy for the condition, and this disease is typically identified at its late stage. Chemotherapy, radiation, and molecular-targeted treatment are typically ineffective against cancer cells. A better grasp of the processes of carcinogenesis, aggressiveness, metastasis, treatment resistance, detection of the illness at an earlier stage, and obtaining a better therapeutic response will be made possible. Researchers have discovered that cancerous mutations mainly affect signaling pathways. The Hippo pathway, as one of the main signaling pathways of a cell, has a unique ability to cause cancer. In order to treat cancer, a complete understanding of the Hippo signaling system will be required. On the other hand, interaction with other pathways like Wnt, TGF-β, AMPK, Notch, JNK, mTOR, and Ras/MAP kinase pathways can contribute to carcinogenesis. Phosphorylation of oncogene YAP and TAZ could lead to leukemogenesis, which this process could be regulated via other signaling pathways. This review article aimed to shed light on how the Hippo pathway interacts with other cellular signaling networks and its functions in leukemia. Hematological malignancies include lymphoma, myeloma, myeloproliferative neoplasms, myelodysplastic syndromes, and leukemia with several subtypes . LeukemiDrosophila mosaic genetic screens first found many mutations in the Hippo signaling pathway [A new signaling pathway, Hippo, has played a crucial role in maintaining organ size by regulating cell proliferation and death in the last decade . Due to  pathway . Because pathway . Mammali pathway .Furthermore, a mutation in the genes encoding Hippo signaling proteins can cause significant organ shape or growth parameters , 12. ForThere is insufficient evidence to identify the tissue specificity and frequency of pathway components and YAP mutations in human leukemias . SeveralThe standard of care for leukemia depends on many factors chosen based on age and overall health, the type of leukemia, and the stage of the disease . Common The human Hippo pathway is based on a kinase signaling cascade including MST1 and MST2, as well as LATS1/2, SAV1 and MOB1, are two types of serine/threonine kinases . When thThe level of YAP1 protein is elevated a variety of cancers, including colorectal cancer (CRC), gastric cancer, esophageal squamous cell cancer (ESCC), human hepatocellular carcinoma (HCC), osteosarcoma –38. The Qu et al. reported that downregulating YAP inhibited cell migration and invasion, and YAP expression level could be a new marker for predicting the prognosis of patients with ESCC . AccordiA wide range of upstream stimuli such as extracellular ligands, organ size, mechanotransduction, environmental stress, energy stress, and cell–cell contact controls YAP/TAZ activation in cancer cells . The actIncreased tissue development has been connected to YAP protein activation, as well as direct target genes like Myc, cell cycle regulators like CycE and E2F1, and apoptosis inhibitors like Diap1 and BIRC3, which have all been identified as contributory factors . Other sIn HSCs, Wnt signaling is critical for maintaining homeostasis , 53. LowThe canonical Wnt pathway is CML’s most seriously impacted Wnt system . BecauseAccording to accumulating evidence, YAP/TAZ, the key effectors in the Hippo signaling cascade, regulate β-catenin levels and activity by physically interacting with β-catenin or Dvl. The first clear evidence that YAP/TAZ inhibited the Wnt/β-catenin pathway came from a study identifying TAZ’s direct interaction with Dvl in the cytoplasm. After Wnt3a stimulation, TAZ knockdown increased Dvl phosphorylation, consequently increasing the nuclear accumulation of β-catenin. Suppressing an upstream kinase in the Hippo pathway improved the connection between TAZ and Dvl, resulting in Wnt/β-catenin pathway downregulation . TAZ’s rBecause Wnt signaling is essential in leukemia stem cells and the microenvironment, targeting Wnt signaling pathways could help treat leukemia . Wnt andThe cellular energy levels, amino acids, and other nutrients affect the rapamycin (mTOR) pathway . Also, iGiven the importance of Hippo and mTOR signaling in growth control, it is not unexpected that links between them have been discovered. Mutation of the tuberous sclerosis complex (TSC), a critical negative regulator of mTORC1, resulted in an mTOR and autophagy-dependent overexpression of YAP proteins in a mouse cancer model . mTORC2 This signaling pathway is critical for transmitting proliferative signals from receptors on membrane-bound . In humaYAP has also been shown to affect how cancer cells respond to inhibitors of the MAPK pathway , 76. TumDeregulation of the Hippo signaling pathway is related to various solid tumors, including lung, breast, liver, and ovary . SignaliMany of the Hippo signaling pathway’s genes have been identified as tumor suppressors, such as MST1/2, SAV1, MOB1a/b, and LATS1/2, whereas others, such as YAP/TAZ, are oncogenes that stimulate malignant cells and allow them to proliferate uncontrollably . SeveralLATS2 gene was significantly overexpressed in patients who suffered AML compared to normal subjects [MST2-ETV6 fusion gene as a core component of the Hippo signaling system, a possible oncogene, in AML patients with t  translocation [YAP and TAZ are functional effectors that regulate gene expression by co-activating various transcription factors involved in leukemogenesis, such as RUNX, TEADS, and SMADS , 85. In subjects   low expression of the ia cells   studies demonstrated that YAP mRNA expression was more significant than healthy controls. In CLL, characterized as lymphoma with B cell accumulation in the blood, bone marrow, and lymph nodes, epigenetic modulation of WWC1 expression was also observed. The WWC1 gene was methylated in around one-third of CLL patients' samples, resulting in lower WWC1 expression . YAP is MicroRNAs are highly involved in the Hippo pathway regulation. Several studies have shed light on the role of the Hippo pathway in tumorigenesis in various types of cancer such as breast, liver, gastric, glioblastoma cancers . ImportaDespite advances in cancer treatment in recent decades, most patients respond poorly after a certain number of treatment cycles, and researchers also face significant challenges in treating cancer . We summSimvastatin also has a potent YAP/TAZ inhibiting action. Ibudilast (a PDE4 selective inhibitor) and Forskolin can promote YAP phosphorylation by preventing cAMP breakdown, implying that PDE inhibitors may be useful in the treatment of cancers with YAP oncogenic activity , 100. DoHippo signaling plays an important role in tumor initiation, invasion, drug resistance, metastatic potential, and self-renewal of cancer stem cells, as well as developmental control. According to studies on this signaling pathway, YAP as a tumor suppressor gene can be involved in many types of cancer. In leukemias, although not much information is available, the increased expression of this protein shows a significant relationship with the poor prognosis of patients. Therefore, the study of the mechanism of action YAP and the factors affecting its inhibition in cancer can be proposed as new pharmacological agents in leukemia treatment.


SignaLink3 contains over 400 000 newly added human protein-protein interactions resulting in a total of 700 000 interactions for Homo sapiens, making it one of the largest integrated signaling network resources. Next to H. sapiens, SignaLink3 is the only current signaling network resource to provide regulatory information for the model species Caenorhabditis elegans and Danio rerio, and the largest resource for Drosophila melanogaster. Compared to previous versions, we have integrated gene expression data as well as subcellular localization of the interactors, therefore uniquely allowing tissue-, or compartment-specific pathway interaction analysis to create more accurate models. Data is freely available for download in widely used formats, including CSV, PSI-MI TAB or SQL.Signaling networks represent the molecular mechanisms controlling a cell's response to various internal or external stimuli. Most currently available signaling databases contain only a part of the complex network of intertwining pathways, leaving out key interactions or processes. Hence, we have developed SignaLink3 ( Signaling pathways describe the chain of molecular events of a cell in response to various external or internal signals. Regulation of many physiological cell functions requires these signaling pathways to work precisely. Deregulation of pathways can lead to disturbances in cellular functions resulting in various diseases. Therefore, it is important to analyse and understand these regulatory processes in order to uncover the workings of these diseases .Traditionally, signaling pathways were considered as a linear chain of events, rather than a complex network of intertwining interactions. However, from a functional viewpoint, to provide a biologically relevant representation of the cell's inner workings we cannot differentiate individual biological processes as independent pathways, as these processes have many common interactors and cross-talks between them . TherefoCurrently available signaling databases use interaction data from high-throughput experiments and text mining, while others such as SIGNOR  and ReacHomo sapiens, we provide signaling data on three main model organisms—Caenorhabditis elegans, Drosophila melanogaster and Danio rerio—in a single web resource. Pathway components are represented at the core of the resource, while their regulatory proteins, transcription factors, miRNAs, and newly added lncRNAs make up each layer of the multi-layered structure. In the update, integrated expression data provides researchers a way to analyze tissue specificity and subcellular localization of signaling networks. SignaLink's new website was developed based on feedback from users to provide an easy-to-use platform with new features and various download formats.As advancements in the field emerge, integrating new data has been a priority in the development of SignaLink. Hence, we now present SignaLink3, the newest version of SignaLink. Next to With the previous version of SignaLink, we have introduced the novel concept of a multi-layered signaling network. We still adhere to this concept in our update due to its efficiency of representing biological processes. The aim of this approach is to represent regulatory mechanisms of signaling pathways as a whole network, rather than separate entities as most other resources do. In the multi-layered structure Figure , the core.g., stimulatory or inhibitory) of the interactions. As a result, of all interactions of SignaLink3 94.3% are directed and 24.2% are signed. In the case of direction prediction, we used methods developed by Liu et al. and Rhodes et al. , TarBase  where ahttps://tflink.net/)). Two major novelties of SignaLink3 is that we doubled the number of miRNAs  and annotated by the pathway they are involved in, as well as their topological features . Each protein is further annotated with their major and minor cellular localization , and tisUsers can access data by navigating to the desired protein's page on the website. The protein page displays an interactive network figure of the interactions of the protein of interest Figure . Here, tThe multi-layered structure of SignaLink not only provides a biologically relevant representation of signaling networks but also caters for customizable download options. On the new download page, users can select any subset of SignaLink, choosing specific species, pathways or layers. With the integrated tissue and subcellular localization data, users can select specific tissues and create a tissue specific network. We also aim to provide a more straightforward way of obtaining data with step-by-step options for customizing the downloaded dataset. SignaLink3 can be downloaded in many widely used formats such as CSV, PSI-MITAB  or BioPAet al. 2020) integrating SignaLink data also indicate the need for such a comprehensive resource. By cross-linking data with such resources, SignaLink's data can reach a wider platform of users. With the rapidly growing amount of data available, one of our priorities is providing an up-to-date resource for our community. Feedback from users - pointing out missing features or personal preferences - has also been considered in the development of SignaLink3. Accordingly, we provide new, previously missing features for an easier and more complete reconstruction of context-dependent signaling networks.SignaLink3 provides an extended and updated resource of signaling networks for humans and three model species. SignaLink3 builds on the previously developed multi-layered network structure and complements it with novel features and annotations to allow creating cell-compartment or tissue-specific networks. SignaLink3 extends signaling pathways by integrating regulatory connections scattered across different databases in one single resource. The derived multi-layered structure presents a more detailed network of signaling events. Since 2013, the community of SignaLink users has grown, reaching researchers in 81 countries with 25,000 visits. Based on the citations on the publication of the previous version of SignaLink , the rese.g., phosphorylation, co-localization, physical association). This additional information allows simulating biochemical reactions and aid in analysis of signal transduction pathways. Another novel feature of SignaLink3 is the possibility of creating tissue specific networks. As pointed out in a review (SignaLink3 is a useful tool for biological modeling, aiding drug discovery research. For these purposes it is crucial to have a thorough annotation of interactions. In SignaLink3, we include predictions for both the direction and effect of interactions, as well as providing information on the type of the interaction (a review  on multia review . Since ta review . HoweverWith the integrated tissue and cellular localization information, and a large number of newly added interactions, SignaLink3 extends signaling pathways with transcriptional, post-transcriptional, post-translational and now lncRNA regulators in four species, making it one of the largest integrated signaling network resources currently available Figure . With a In further developments, we aim to include emerging analysis methods and computational approaches in future versions of SignaLink. Our automated pipeline makes it simple to integrate novel data to always provide an up-to-date source of signaling network data with bi-annual data updates. With our growing community, we plan to prioritize user feedback to make SignaLink fit the needs of researchers. As the prevalence of epigenetics research rises, we also plan to integrate epigenetics information, such as methylation profiles or chromatin regulation data to future updates of SignaLink. Currently, transcription factor - target gene interactions mostly consider transcription factor binding sites in the promoter regions of the target genes. In future updates we plan to further extend these binding sites to more distant enhancer regions as well. With the introduction of the option of creating tissue-specific networks we made a leading step in signaling network research. However, with the emergence of single-cell sequencing technologies we plan to provide a way to create cell-type specific networks with further developments.http://SignaLink.org.SignaLink data is freely available for download in widely used formats, including CSV, PSI-MI TAB or SQL at gkab909_Supplemental_FileClick here for additional data file.


Rupicapra rupicapra and R. pyrenaica) are currently recognized by taxonomy and further subdivided into seven and three subspecies, respectively. However, recent research based on molecular markers finds this classification questionable. We aim to increase the resolution of published research on chamois phylogeny by including mitogenomes of all available subspecies, including the previously unpublished mitogenomes of R. r. balcanica and R. r. tatrica subspecies. The inferred phylogeny based on the full mitogenomes confirms the previously reported genus subdivision in three clades and its monophyletic positioning within the Caprinae. Phylogeny and taxonomy of Rupicapra species thus remain controversial prompting for the inclusion of archeological remains to solve the controversy.Two species of chamois (Rupicapra rupicapra and R. pyrenaica) are currently classified as least-concern by the IUCN , inconsistencies on the subspecies classification reported in literature make it challenging to assess the conservation status of the single subspecies. Previous studies relying on mitochondrial genes, sometimes in combination with nuclear or Y-chromosome markers, reported the presence of clusters corresponding to the geographic distribution but highlighting ambiguities in the genus phylogeny. Here we report novel de novo assembled sequences of the mitochondrial genome from nine individuals, including previously unpublished R. r. balcanica and R. r. tatrica subspecies, and use them to untangle the genus phylogeny. Our results based on the full mitogenome inferred phylogeny confirm the previously reported genus subdivision in three clades and its monophyletic positioning within the Caprinae. Phylogeny and taxonomy of Rupicapra species thus remain controversial prompting for the inclusion of archeological remains to solve the controversy.Although the two species of chamois ( Rupicapra, is the most abundant mountain-dwelling ungulate in Europe and the Near East, and is currently recognized to be divided into two species: Rupicapra rupicapra (Northern chamois) and Rupicapra pyrenaica (Southern chamois), further subdivided into seven  and three  subspecies, respectively [The chamois, genus ectively ,2. The cectively ,4. Althoectively  and refeb (cytb) identified three lineages geographically separated, a western clade in the Western Alps and Iberia, a central clade in the Apennines and the Chartreuse Massif, and an eastern clade covering all regions east of the Alps [Rupicapra mitogenomes are available. Hassanin et al. [Caprini tribe but were unable to resolve the relationships of Rupicapra and a few other genera. In their study, the two Rupicapra species formed a separate branch in the tree [Previous studies relying on cytochrome the Alps . This suthe Alps , differsthe Alps ,8,9. Thethe Alps ,8,9, as the Alps . Howeverthe Alps ,11,12, wthe Alps ,14, thusthe Alps ,16. So fn et al.  were thethe tree  but reprthe tree  confirmeR. r. rupicapra and R. r. balcanica were introduced for hunting purposes and led to a molecularly identified hybrid population [To further complicate the picture, reintroductions and translocations—if performed without previous knowledge on the involved individuals’ taxonomic status ,20—mightpulation . HoweverCaprini, in a sister-group relationship with Ammotragus and Arabitragus, while prompting for additional multi-marker studies also involving historical samples to disentangle the genus phylogeny and taxonomy.In this study, we aimed to contribute to the unraveling of the complex genus phylogeny by increasing the number of individuals and subspecies investigated using the whole mitochondrial genome. Our results confirmed the grouping of individuals from Mt. Velebit with the Balkan subspecies, the subdivision of the genus into three mitochondrial clades and its monophyletic positioning within the Rupicapra subspecies . All samples were collected from dead animals during regular hunting seasons. Genomic DNA was extracted from tissue samples using a standard phenol-chloroform method. DNA fragments were then treated according to the Illumina DNA sample preparation protocol. Genome sequencing was performed on the Illumina HiSeq 2500 platform with paired-end adaptors and 100 bp reads length for libraries of 350 bp inserts. To investigate the phylogenetic relationships, we downloaded one mitogenome sequence for each of the 34 (sub)species from the subfamily Caprinae available in GenBank and five outgroups: Damaliscus pygarus, Damaliscus lunatus, Bos taurus, Bubalus bubalis, and Muntiacus reevesi (We collected samples from  reevesi .R. rupicapra sequence (FY207539) as seed. The resulting circular genomes were verified as belonging to the Genus in BLAST [R. rupicapra sequence using MEGA X [Raw sequences were corrected using the Pollux program  with defin BLAST . Obtaineg MEGA X , Bioeditg MEGA X , and BWAg MEGA X  softwareg MEGA X .Caprini [Rupicapra subspecies. The ND6 region was reverse complemented to present the same reading direction as the other protein-coding sequences [Newly assembled mitogenomes that passed the quality controls  were aliCaprini  were excCaprini  was usedCaprini , regionsCaprini  was usedequences . DNAsp wequences  with 100Caprinae species were reconstructed using a reduced alignment containing a single sequence per Rupicapra subspecies, mitogenome sequences of Caprinae species available in GenBank, and five Bovidae as outgroups. The complete dataset used for this analysis consisted of 45 mitogenome sequences. Relationships within Rupicapra were investigated using all Rupicapra sequences available in GenBank (n = 5) and the nine new sequences obtained within this research. Mitogenome sequences of Ammotragus lervia and Arabitragus jakari were included as outgroups. PartitionFinder2 [Caprinae and Rupicapra datasets was computed with IQ-TREE default parameters, genetic code set to vertebrate mitochondrial, substitution model TIM2+I+G4, and ultrafast bootstrap analysis UFBoot2 [Phylogenetic relationships among nFinder2  was usednFinder2  was usednFinder2  and BayenFinder2 . Stochas UFBoot2 .Caprinae was performed with GTR+I+G4 model and Markov chain Monte Carlo simulation for 1.2 million generations. The trees were sampled every 500 generations until reaching an average standard deviation of split frequencies of 0.011. MrBayes parameters for Rupicapra were GTR+G4 and 1.7 million generations, sampled every 200 generations until reaching an average standard deviation of split frequencies of 0.006. For both analyses, the convergence of chains was additionally checked by verifying the stationarity of the log probability graphs [Bayesian inference on the y graphs . For eacRupicapra subspecies were calculated in Mega X using the Maximum Composite Likelihood model [Tree editing and visualization were performed using the R package ggtree . Within od model , rate vaod model  and inclRupicapra, which showed the typical organization  and nucleotide  diversity, with high variability between the two species and among subspecies .The 14 bspecies . A similceptions . Howeverceptions . Althougceptions , the pS-Damaliscus and the Ovicaprini, the basal divergence of Pantholops within the Ovicaprini, and a well-supported separation of the Ovibovini. Within the Caprini tribe, Capra and Hemitragus form a goat-like clade and the more distant Pseudois and Budorcas as sister groups; Ovis and Oreamnos group together in a sheep-like clade; Ammotragus and Arabitragus showed a sister group relationship and finally, Rupicapra was monophyletic  and the region analyzed  [R. rupicapra and R. pyrenaica) and three mitochondrial clades corresponding to the geographic distribution of the populations .The phylogeny and taxonomy of  complex ,9. Initi complex . Later s complex ,6,7,18, ogenome) ,18. CurrCaprinae tree  was observed compared to the Iberian chamois (0.012), and unexpectedly high variation at the ND6 gene . GC content was more homogeneous between the two species, with a single noticeable difference at the ATP8 gene, with the Alpine chamois showing lower variability . While estimates of nucleotide diversity on small samples are found to be underestimated [Quite interestingly, Pérez and colleagues  reportedstimated , it is iamois 0.0, and unestimated ,55,56. Istimated , epilepsstimated , and musstimated , whereasstimated , epilepsstimated  and cancstimated , making Overall, our results provide increased resolution compared to the available literature, confirm the phylogeny previously reported for both whole and partial mitochondrial sequences and the discrepancy among markers in phylogenetic reconstruction of the genus. Pérez and colleagues  explaine


Human T-lymphotropic virus 1 and 2 (HTLV-1/2) belong to the delta group of retroviruses which may cause a life-long infection in humans, HTLV-1 leading to adult T-cell leukemia/lymphoma and other diseases. Different transmission modes have been described, such as breastfeeding, and, as for other blood-borne pathogens, unsafe sexual activity, intravenous drug usage, and blood transfusion and transplantation. The present systematic review was conducted to identify all peer-reviewed studies concerning the work-related infection by HTLV-1/2. A literature search was conducted from January to May 2021, according to the PRISMA methodology, selecting 29 studies: seven related to health care workers (HCWs), five to non-HCWs, and 17 to sex workers (SWs). The findings showed no clear evidence as to the possibility of HTLV-1/2 occupational transmission in HCWs, according to the limited number and quality of the papers. Moreover, non-HCWs showed a higher prevalence in jobs consistent with a lower socioeconomic status or that could represent a familial cluster, and an increased risk of zoonotic transmission from STLV-1-infected non-human primates has been observed in African hunters. Finally, a general increase of HTLV-1 infection was observed in SWs, whereas only one paper described an increased prevalence for HTLV-2, supporting the urgent need for prevention and control measures, including screening, diagnosis, and treatment of HTLV-1/2, to be offered routinely as part of a comprehensive approach to decrease the impact of sexually transmitted diseases in SWs. Human T-lymphotropic virus 1 and 2 (HTLV-1 and HTLV-2) belong to the delta group of retroviruses, together with virus 3 and 4 , their simian counterparts , and the bovine leukemia virus (BLV) [It has been estimated that 5–10 million people are currently infected by HTLV-1, mainly in the endemic regions, including South Japan, Northeastern Iran, sub-Saharan Africa, almost all of the Caribbean islands, southeastern U.S. regions, Melanesia, and South America ,3. HTLV-Most HTLV-1-infected subjects are asymptomatic, but 2–5% suffer the onset of adult T-cell leukemia/lymphoma (ATL), while a smaller proportion of infected people develops a neurological disorder called HTLV-1-related myelopathy/tropical spastic paraparesis (HAM), or else other chronic illnesses, such as uveitis and dermatitis ,7. UnlikHTLV-1 exhibits little genetic heterogeneity, being spread by the clonal division of infected cells, rather than by reverse transcription-mediated viral replication. Seven different genomic species (A-G) have been identified, mostly related to the geographic origin; subtypes A, B, and C are known to cause ATL, HAM, and dermatitis ,11,12. STransmission of HTLV-1 is lymphocyte-mediated and requires cell-to-cell contact . Severaln = 42 selected items.A systematic literature search was conducted to select available information according to the PRISMA  methodology, to identify all peer-reviewed studies of the relationship between HTLV-1/2 infection and the occupational setting. Firstly, a systematic review of the literature was made using the PubMed, Scopus, Web of Science databases. A search strategy was developed and adapted for each database, and then one researcher performed data extraction from January to May 2021. The variables to be analyzed were selected according to the population, exposure, comparison, and outcome (PECO) method, which is often used for systematic review and is easily adapted to a particular case and combined with other essential variables. Characteristics of individuals (population), exposure to the phenomenon (direction), and their consequences (outcome) were considered. The following keywords: (“HTLV-1 OR HTLV-2) AND  were used for the search. Screening of resources took place in the first phase, by reading the articles titles and abstracts and removing duplicates . This phn = 29 items was made.In the second stage, the full texts of the selected articles were screened independently by two co-authors, based on the inclusion criteria, namely, the English language, case reports or research article, topic on HTLV-1/2 infection or anti-HTLV-1/2 seroprevalence and occupational setting, and exclusion criteria, that were non-occupational setting studies or topics on other viruses. To establish a final set of papers, the two reviewers compared their findings to identify consistencies and discrepancies. Any differences were resolved by mutual agreement. At the end of this second phase, the final selection of The selection for papers on SWs was made according to the World Health Organization (WHO) definition, that considers women, men, and transgendered SWs as those people who receive money or goods in exchange for sexual services, and who consciously define those activities as income-generating, even if they do not consider sex work as their occupation .The details of the articles included in the review are reported according to the occupational setting, distinguishing HCWs, non-HCW and SWs. Seven articles on HTLV-1/2 infection in HCWs were collected, including five observational studies and two case-report studies .The study performed by Hewagama et al.  retrospeIn a survey performed from 2000 to 2008 to investigate the epidemiology of HTLV-1/2 in US Armed Force military personnel in order to ensure emergency transfusion safety, a higher rate of HTLV-1/2 diagnosis was found in military HCWs  vs. combat military personnel  and other military personnel , yielding a relative risk (RR) for HCWs vs. combat personnel of 2.54  .Stuver et al.  observedA seroprevalence survey was conducted by Goubau et al.  among 42A HTLV-1 seroprevalence study, including 291 intravenous drug abusers, 45 household contacts of surveyed drug abusers, and 39 laboratory HCWs at a methadone maintenance center in Rome , was conducted between 1985 and 1987 . SerologBarreto  reportedA case of HTLV-1 associated to a HAM was described by Goubau et al.  in a 57-Five observational studies on work-related HTLV-1/2 prevalence in non-healthcare occupational settings were collected and included in the review .p < 0.001). All the HTLV-1-positive hunters bitten by a gorilla or chimpanzee were infected by a subtype B strain, like that in monkeys in the same area, whereas two hunters attacked by small monkeys were infected by subtype F, that was very close to the strains found in those small monkeys. Mother-to-child infection was excluded in the six infected subjects in whom transmission could be investigated, clearly indicating that hunting NHPs should be considered an occupational activity at risk of zoonotic transmission by STLV-1.A cross-sectional survey made in 2005–2012 compared the HTLV-1 prevalence in 269 individuals  from rural villages and settlements in Cameroon (exposed), bitten by NHPs mostly during hunting activities, vs. 269 matched subjects from the same settlers, but referring no previous NHPs attacks (controls) . The stuCercopithecus nictitans resulted in infection by a subtype D strain that was closely related to the STLV-1 that infected this monkey species, while her mother was infected by a subtype B strain. The observed results, therefore, confirmed that hunters in Africa should be considered a high-risk population, because a severe NHPs bite is a risk factor for STLV-1 acquisition.A similar cross-sectional study was performed by Kazanji et al.  in southFew other studies have investigated work-related HTLV-1/2 infection in non-HCWs. No cases of HTLV-1 infection were assessed in 28 Mexican slaughterhouse workers in a survey performed to rule out cross-reactivity with BLV . NorrgreSeventeen articles related to the SWs were collected, including 16 observational studies and one case report .A cross-sectional study was conducted from 2005 to 2006 in four cities in the State of Parà (Brazil) amongst 339 female SWs, to analyze sociodemographic characteristics, sexual habits, frequency, circulating molecular subtypes of HIV-1 and the HTLV-1/2 seroprevalence [A recent study by Paulino-Ramirez et al.  in the pA cross-sectional study investigated HTLV-1/2 co-infection, between April 2015 and December 2017, in the 21 female SWs showing HBV infection among a total group of 153 female SWs from the Marajo Archipelago (Northern Brazil) . No HTLVp < 0.01), no cases of HTLV-1 being detected among the 224 SWs born after 1979. Among the investigated variables, age, earlier birth cohort, native home in the Andes, duration of sex work, age at the time of starting sex work, as well as HIV seropositivity, were positively associated with HTLV-1 infection.Data collected in 1938 female SWs from Callao (Perù), screened for HTLV-1 infection over three study periods between 1993 and 2010, were used to examine the trend in HTLV-1 prevalence . The oveThe report by Bautista et al.  investigThe prevalence of HTLV-1/2 infection amongst 166 female SWs, 120 pregnant women (PW) and 78 female secondary school students from Ibadan (South-Western Nigeria) was assessed by Forbi et al. . The oveBerini et al.  studied Another cross-sectional study was conducted between March 2000 and March 2002 by Pando et al.  among 61A survey conducted by Zehender et al.  between Trujillo et al.  reportedBetween 1993 and 1996, Chen et al.  studied An epidemiologic study was conducted by Zurita et al.  to deterA cross-sectional survey of the HTLV-1/2 seroprevalence was performed in North-East Brazil, in the period July 1993–February 1994 in 496 female and 171 male SWs, 814 PW, 494 TB and 395 STD patients, 427 prisoners . The preA cross-sectional observational study by Bellei et al.  evaluateIn a cross-section observational study, Zapata-Benavides et al.  performeYoshida et al.  investigA case-report of HTLV-1 infection was described by Caterino-De-Araujo et al.  in a 25 A comparison between the prevalence of HTLV-1/2, HIV-1 and other STDs in SWs, has been described in 11 of the 16 observational studies included in the review .The present review investigated the relationship between different working activities and infection by HTLV-1/2 to probe whether prevention in workplaces should play a role in the global approach to the prevention of the diseases caused by HTLV-1/2.Previous studies have often considered HCWs at higher risk of HTLV-1/2 infection, according to the possible virus transmission by the parenteral route following biological accident . Our sysA possible explanation for the different behavior between HTLV-1/2 and the other blood-borne viruses could be related to the fact that HTLV-1/2 are cell-associated viruses, that probably require the transfer of whole cells for virus transmission to occur. In fact, unlike other retroviruses, lymphocytes infection by free virions is very inefficient, as it is for most cell types except dendritic cells, while transmission is greatly improved upon the establishment of cell-cell contact . A minimThe prevalence of HTLV-1/2 infection in occupational groups other than HCWs was investigated in few cross-sectional studies, most of which were conducted in endemic areas, over a period ranging from 1990 to 2015. Firstly, a higher HTLV-1 prevalence was observed in Central Africa hunters, where interspecies transmission of STLV-1 by NHPs has been considered as a possible source of infection. Molecular characterization of primate T-lymphotropic viruses, both simians and humans, reveals that some HTLV subtypes share closer genetic ties to certain STLVs than to other HTLV subtypes, suggesting sustained zoonotic transmission of STLV between NHPs and humans . PhylogeAmong the reviewed studies, a higher rate of HTLV-1 seropositivity was observed in farmers, laborers and skilled tradesman in a Jamaican population, and in fishing, livestock and forestry workers in a high prevalence Japanese area ,32. HoweOur literature review showed a general increase of HTLV-1 infection among SWs, whereas only one paper described an increased prevalence for HTLV-2, with all the studies, except that by Forbi et al., including confirmatory tests in the analytical procedure . OverallThe high prevalence of HTLV-1 observed in SWs confirmed the epidemiologic importance of sexual transmission, suggesting the essential role of SWs in maintaining the infection and transmission to the general population. The mechanism of transmission in the context of sexual intercourse explains why transmission is more effective from men to women. HTLV-1/2 transmission, in fact, is related to infected T-cell lymphocytes present in the semen that come in contact with the female genital mucosa, mainly composed of epithelial cells which can capture HTLV-1/2 at the apical face and release them at the basal face through transcytosis. HTLV-1 could then infect dendritic cells underneath the epithelial barrier . Vice veAnalysis of the variables influencing the HTLV-1/2 prevalence showed higher rates of seropositivity among SWs who referred to a history of blood transfusions or illicit drug use, and in those reporting low condom use ,42. MoreTo the best of our knowledge, this is the first systematic literature review to analyze the risk of work-related infection by HTLV-1/2. Although a higher risk of infection has been supposed for HCWs, our review did not identify a clear increased rate of seropositivity in HCWs than in the general population, also due to the severe limitations of the observational studies, whereas the main evidence of infection was referred to case-reports in which inadequate use of personal protective equipment had been reported. Definite conclusions, therefore, are not possible on the basis of the actual knowledge and further studies, particularly focusing on HCWs with history of exposure to biological fluids of HTLV infected patients, are needed to clarify the real extent of the occupational risk for HCWs. An increased risk of zoonotic transmission has been observed in Central Africa hunters who perform their activities in STLV-1 infected NHPs habitats, but epidemiological evidence in larger populations is still scarce. Finally, our review confirms that SWs have a higher risk of contracting mainly HTLV-1, although the limit of the large variability observed in HTLV-1 prevalence, supporting the urgent need for prevention and control measures, including screening, diagnosis and treatment of HTLV-1/2, to be offered routinely as part of a comprehensive approach to decrease the impact of STDs in SWs . Neglect


Antidepressants are known to show heterogeneous effects across individuals and conditions, posing challenges to understanding their efficacy in mental health treatment. Social media platforms enable individuals to share their day-to-day concerns with others and thereby can function as unobtrusive, large-scale, and naturalistic data sources to study the longitudinal behavior of individuals taking antidepressants.We aim to understand the side effects of antidepressants from naturalistic expressions of individuals on social media.On a large-scale Twitter data set of individuals who self-reported using antidepressants, a quasi-experimental study using unsupervised language analysis was conducted to extract keywords that distinguish individuals who improved and who did not improve following the use of antidepressants. The net data set consists of over 8 million Twitter posts made by over 300,000 users in a 4-year period between January 1, 2014, and February 15, 2018.Five major side effects of antidepressants were studied: sleep, weight, eating, pain, and sexual issues. Social media language revealed keywords related to these side effects. In particular, antidepressants were found to show a spectrum of effects from decrease to increase in each of these side effects.This work enhances the understanding of the side effects of antidepressants by identifying distinct linguistic markers in the longitudinal social media data of individuals showing the most and least improvement following the self-reported intake of antidepressants. One implication of this work concerns the potential of social media data as an effective means to support digital pharmacovigilance and digital therapeutics. These results can inform clinicians in tailoring their discussion and assessment of side effects and inform patients about what to potentially expect and what may or may not be within the realm of normal aftereffects of antidepressants. As mental health concerns continue to surge as an epidemic, arguably exacerbated in the present and the near future due to the ongoing COVID-19 pandemic ,3. HowevThis short paper targets the above gap by adopting an observational study approach using natural language and machine learning methodologies on large-scale social media data. Our work draws motivation from the success of using social media as an effective source of unobtrusive, real-time, low-cost, and naturalistic data to infer wellbeing. Social media platforms enable individuals to share and express their day-to-day psychosocial concerns; therefore, this is a form of longitudinal verbal and behavioral data, and computational linguistic approaches have helped reveal naturalistic patterns of mood, behavior, cognition, psychological state, and social milieu of individuals and collectives -15.Linguistic cues and social interactions on social media have enabled the inference of psychopathologies including depression, anxiety, stress, suicidal ideation, and loneliness ,12,16-19This work uses natural language and causal inference analytic techniques on self-initiated social media expressions of antidepressant users to study the heterogeneity of individual and drug-specific outcomes. Our rationale is situated in the emergent body of empirical evidence in “digital pharmacovigilance” . DigitalTo begin with, a list of Food and Drug Administration–approved antidepressants and antidepressant augmentation drugs was developed in consultation with the psychiatrist coauthor (JT). This list included 297 brand names mapped to 49 generic names across the four major drug families: serotonin-norepinephrine reuptake inhibitors (SNRIs), selective serotonin reuptake inhibitors (SSRIs), tricyclic antidepressants, and tetracylic antidepressants. Next, the Twitter application programming interface was queried for public English posts containing the brand name or generic name of these drugs between January 1, 2015, and December 21, 2016, to obtain 601,134 posts by 230,573 unique users. Because a mention of a drug does not necessarily indicate a self-intake, a personal medication intake classifier built in prior work  was usedagomelatine, amineptine, amitriptyline, amoxapine, bupropion, butriptyline, citalopram, clomipramine, desipramine, desvenlafaxine, dibenzepin, dosulepin, doxepin, duloxetine, escitalopram, etoperidone, fluoxetine, fluvoxamine, hydroxynefazodone, imipramine, iprindole, levomilnacipran, lofepramine, maprotiline, mazindol, meta-chlorophenylpiperazine, mianserin, mirtazapine, nefazodone, nisoxetine, nomifensine, norclomipramine, northiaden, nortriptyline, opipramol, oxaprotiline, paroxetine, protriptyline, reboxetine, sertraline, setiptiline, trazodone, triazoledione, trimipramine, venlafaxine, vilazodone, viloxazine, vortioxetine, zimelidineThis work aimed to obtain individuals who most and least improved in mental health symptomatic outcomes after the use of antidepressants. Replicating prior work , the stun-grams in the posttreatment data sets of treatment users who showed contrasting changes in symptomatic outcome measures of depression, anxiety, stress, suicidal ideation, and psychosis. A detailed approach is included in the supplementary information in To drill into the posttreatment linguistic markers associated with the symptomatic changes per medication, an unsupervised language modeling approach, the Sparse Additive Generative (SAGE) model , was empTo understand drug-specific effects, first, the posttreatment linguistic markers associated with the symptomatic outcomes of individuals who used antidepressants were compared . Table 1Antidepressants are known to affect the circadian rhythms . Sleep-rChange in weight is a prominent behavioral side effect of many psychiatric medications . The keyKeywords related to eating occurred saliently in both the most and least improved users. For instance, keywords such as “eating disorder” and “hungry” dominated in the posts by the improved users of fluoxetine, whereas “want eat” dominated in the least improved users of sertraline, and “food” dominated in the least improved users of duloxetine. Neuropathic pain is a symptom in depression  and is aAntidepressants are likely to affect sexual issues . In thisSummarily, our linguistic analysis reveals many keywords potentially related to prominent side effects of the antidepressants. There is a lack of simple and systematic means to understand side effects of antidepressants ,5. It isOur knowledge about the etiopathogenesis of mental diseases continues to be “top to bottom” instead of being “bottom up” . ConsideFinally, this study adopts a quasi-experimental study design, which cannot establish a “true causality.” However, it is more robust than more simple correlational analysis because of minimizing the confounders. This work motivates future computational study designs that combine the power of complementary digital data sources, including social media, and other ubiquitous data streams to obtain a more holistic understanding of an individual’s behavior with respect to the use of antidepressants.


Given the heterogeneity of endosomes and lysosomes, due to their specific protein and lipid composition, subcellular localization, and membrane shape, it is not surprising they have also arisen as dynamic signaling platforms. Endosomes and lysosomes elicit an array of signaling events, including regulation of gene expression, cell survival, cytoskeletal organization, nutrient uptake, lipid transport, and autophagy , followed by their internalization by endocytosis. These cargoes are then transported utophagy . These outophagy .In this research topic, we assembled review and original research articles that provide recent advances of molecular mechanisms that are a signature for endosomal and lysosomal function under physiological and pathological conditions. We envision that these articles will illustrate a thorough overview of the current progress and future directions relying on proposed mechanistic models.Kervin et al. present an elegant meta-analysis that demonstrates that binding of PX domain-containing proteins to membranes is regulated by modification on lysine and arginine residues of PX domains. These side chain modifications, such as acetylation, methylation, malonylation, succinylation, hydroxysobutyrylation, and glycation, neutralize the positive charges on lysine and arginine residues, thus, impairing electrostatic interactions with phosphoinositides. These modifications on PX domains, named MET-stops, are also predicted to interfere with “hot spot” sequences for membrane association as identified by the Membrane Optimal Docking Area algorithm. Therefore, MET-stops represent erasers of protein membrane targeting.Peripheral signaling and trafficking proteins target endosomal and lysosomal compartments through interactions with phosphoinositides. One of the best studied phosphoinositide-binding domains is the PX domain, which displays a wide range of specificities for these lipids. Roach et al. provide a detailed review on the function of the endosomal ESCRT protein TOM1 in physiological and disease scenarios. TOM1 participates in cargo trafficking in coordination with other ESCRT proteins and phosphoinositides. TOM1 is also involved in other cellular functions, including autophagy, immune responses, neurodegeneration, and cancer, although many questions regarding molecular interactions in these processes and crosstalk with TOM1’s endosomal function still remain unanswered. Recently, TOM1 has been shown to be hijacked under Shigella flexneri infection following accumulation of intracellular phosphatidylinositol 5-phosphate, in which the ESCRT function of TOM1 is impaired. We hope that more research will shed light on the molecular mechanisms governing TOM1 function in these processes.Cargo trafficking relies on the function of the cargo sorting machinery known as ESCRT . Roach eSchmidt-Arras and Rose-John review the complex spatial regulation of signaling in response to the pro-inflammatory cytokine interleukin 6 (IL-6). The IL-6 receptor consists of the IL-6-binding subunit α and the signaling subunit gp130, which is shared among many different cytokines. The trafficking of the receptors to endosomes and lysosomes is an important layer of regulation. Following endocytosis of the IL-6 receptor-IL-6 complex, signaling can continue from within the lumen of endosomal compartments. Moreover, the receptor can be recycled to the plasma membrane, but also targeted to lysosomes for its degradation. As summarized by Schmidt-Arras and Rose-John, mutations that disrupt endo/lysosomal IL-6 receptor trafficking are a cause for disease.Dabral and van den Bogaart review the roles of phospholipase 2 (PLA2) in phagocytosis. Phagocytosis is mechanistically related to receptor-mediated endocytosis, and is the process by which immune cells, such as macrophages and neutrophils, ingest pathogens and subsequently kill them in phago-lysosomes. PLA2 is a family of enzymes that hydrolyzes phospholipids, producing a free fatty acid and a lysolipid. In addition to the canonical role of PLA2 in eicosanoid signaling  clusters lysosomes. FUS is an RNA-binding protein involved in regulation of transcription, RNA splicing, and RNA transport , which uses ATP hydrolysis to drive the translocation of hydrogen ions across endosomal and lysosomal membranes resulting in a reduction in lumenal pH. Signaling Proteins for Endosomal and Lysosomal Function provides a more comprehensive understanding of several aspects of the endosomal and lysosomal functions, which will serve as a platform for novel discoveries in these intriguing organelles.In summary, this collection of articles for the Research Topic


Crassostrea gigas) variety “Haida No.1” as the material with which to unravel the role of IIS systems in growth regulation in oysters. Systematic bioinformatics analyses allowed us to identify major components of the IIS signaling pathway and insulin-like peptide receptor (ILPR)-mediated signaling pathways, including PI3K-AKT, RAS-MAPK, and TOR, in C. gigas. The expression levels of the major genes in IIS and its downstream signaling pathways were significantly higher in “Haida No.1” than in wild oysters, suggesting their involvement in the growth regulation of C. gigas. The expression profiles of IIS and its downstream signaling pathway genes were significantly altered by nutrient abundance and culture temperature. These results suggest that the IIS signaling pathway coupled with the ILPR-mediated signaling pathways orchestrate the regulation of energy metabolism to control growth in Pacific oysters.The involvement of insulin/insulin-like growth factor signaling (IIS) pathways in the growth regulation of marine invertebrates remains largely unexplored. In this study, we used a fast-growing Pacific oyster ( Nutrient abundance is one of the most important environmental factors that are critical to the growth and reproduction of all organisms. Nutrient sensing and energy metabolism are coordinated by networks of signaling cascades, such as the insulin/insulin-like growth factor signaling (IIS), target of rapamycin (TOR), and adenosine monophosphate-activated protein kinase (AMPK) signaling pathways . The IISAs one of the most important nutrient sensors, the mechanism of the IIS pathway in the growth regulation of vertebrates has been well studied. The IIS signaling pathway—including insulin/insulin-like growth factors (IGFs), insulin/IGF receptors , insulin receptor substrate (IRS), insulin-like growth factor-binding proteins (IGFBPs), and insulin-like growth factor-binding protein complex acid labile subunits (IGFALSs)—regulates multiple cellular processes, including cell growth and proliferation, energy metabolism, hormone secretion, and glucose homeostasis ,4,5. In Only insulin-like peptides, but no growth hormone, have been identified in various invertebrates ,12,13. TCrassostrea gigas) is one of the most widely cultured marine mollusks, and has been introduced from Asia to many other countries around the world [C. gigas in China performed for over ten years has produced a fast-growing variety named “Haida No.1”, which exhibits significant growth advantages over wild oysters [C. gigas, and showed that expression levels of ILPs were significantly higher in fast-growing “Haida No.1” than in wild oysters [ILP genes and on the growth of the oysters. To further understand the role of the IIS signaling pathway in the growth regulation of oysters, in this work, we performed extensive multi-omics data mining in order to identify the key components of the IIS, PI3K-AKT, and RAS-MAPK signaling pathways in C. gigas. The expression profiles of the major genes of the IIS and ILPR-mediated signaling pathways were determined in “Haida No.1” and in wild oysters. Fasting/re-feeding and low-temperature culture experiments were performed in order to further understand how the IIS signaling pathway was altered by nutrient levels and temperature to affect the growth of C. gigas. This work provides a systemic characterization of the IIS and ILPR-mediated signaling pathways involved in the growth of C. gigas, which will provide valuable information toward our understanding of the molecular basis of growth regulation in oysters and other invertebrates.The Pacific oyster  and the IGFBPRP from other mollusks, while the other six vertebrate-specific IGFBP members were clustered into other separate clades  signaling pathways in regulating growth and energy metabolism has been well-studied in vertebrates, while the role of IIS pathways in the growth regulation of marine invertebrates, such as oysters, remains largely unexplored. In a previous study, we identified four insulin-like peptide genes in C. gigas suggested a limited role for the IIS in growth and metabolism regulation. Phylogenetic analysis showed that the sole C. gigas ILPR was clustered with the ILPRs from other invertebrates, and had a relatively closer homology with the insulin receptor (INSR) in comparison with other members of the IR subfamily in vertebrates—including the IGF1R, IGF2R, and INSRR—indicating the similar physiological functions of the ILPR in C. gigas and the insulin receptor in vertebrates. Notably, although four insulin-like peptide genes were found in C. gigas, there was only one ILPR, which was strikingly different from the case in vertebrates, deserving further investigations on the evolution of ligand and receptor binding in insulin-like peptides. The IRS, which could bind to the phosphorylated tyrosine residues of beta subunits of the ILPR, is involved in transducing the IIS signaling pathway within the cell [C. gigas, indicating specific binding of the IRS to beta subunits of ILPR in C. gigas. Identification of the IIS, PI3K-AKT, and RAS-MAPK signaling pathway genes in the cell ,42. In oC. gigas, and the IGFBPRP was clustered into one clade with the IGFBPRPs from other mollusks and the IGFBP7 from vertebrates. Our analysis suggested that the C. gigas IGFBPRP had a similar evolutionary origin and physiological function to the IGFBP7 in vertebrates, supporting the idea that the IGFBPRP (IGFBP7) may be the most ancient gene among the IGFBP subfamily. Furthermore, the number of ALSs is only one or two in vertebrates, while there are usually five to seven IGFALSs in arthropods and mollusks. The obviously larger number of IGFALSs in invertebrates than in vertebrates indicates that some of the ALSs could have been lost during evolution after the divergence of invertebrates and vertebrates.The IGFBP family is composed of 6 distinct types of IGFBP (IGFBP1–6) and 10 IGFBPRPs in vertebrates, which is evolutionarily ancient and conserved . In inveC. gigas, as revealed in our previous study. In the present work, the expression profiles of the major genes in the IIS signaling pathway were further determined in order to confirm the systemic regulatory role of the IIS signaling pathway in the growth regulation of oysters. Our results showed that the major genes of the IIS signaling pathway, including ILPR, IRS, and IGFBPRP, were all expressed higher in the fast-growing “Haida No.1” than in wild oysters. However, the C. gigas ALSs showed distinct expression profiles. The ALSs have been reported to play an indispensable role in the growth regulation of vertebrates through the GH/IGF system [ALS5594 gene was expressed higher in wild oysters, while the ALS7789 and ALS7489 genes were expressed at higher levels in the “Haida No.1” oysters, indicating that the ALS also played important but diverse roles in the growth regulation of C. gigas. Higher expression levels of insulin-like peptide genes were associated with fast growth in F system ,45,46. IPI3K, PDK, AKT, RAS, MEK, ERK, TOR, and GSK3β genes, were all expressed at higher levels in the “Haida No.1” oysters, while the negative regulators PTEN, FoxO, and S6K were all highly expressed in wild oysters. Therefore, ILPR is speculated to transduce signaling from the active ternary complex “ILP–IGFBP–ALS” to the downstream PI3K-AKT and RAS-MAPK signaling pathways, functioning to regulate energy metabolism and cell proliferation in oysters  gene in our previous study [C. gigas. In addition, the expression of the ALS7789 and ALS4336 genes was upregulated during the fasting and re-feeding process, which is consistent with the expression profiles of the MIRP3, MIRP3-like, and ILP7 genes in our previous study [ALS7789 and ALS4336 genes may be associated with the physiological role of molluscan insulin-related peptide 3 (MIRP3), molluscan insulin-related peptide 3-like (MIRP3-like), and insulin-like peptide 7 (ILP7), and are indispensable in energy metabolism under the control of neuroendocrine activity. Furthermore, low temperature greatly suppressed the expression of the ILPR, IRS, IGFBP, ALS7789, ALS7489, and ALS1089 genes. These results suggest that the IIS signaling pathway is responsive to fluctuations of nutrition condition and ambient temperature. Fasting/re-feeding and low-temperature culture experiments were performed in order to further confirm the hypothesis. In our present study, the major genes of the IIS signaling pathway—including the us study . These rus study . We specTOR and S6K genes was decreased during fasting, and recovered to normal levels after re-feeding. GSK3β phosphorylates the serine and threonine sites of a variety of substrates, including glycogen synthase, in order to regulate glycogen synthesis [GSK3β was inhibited under food deprivation conditions, and recovered to normal levels once food was abundant, indicating the reduction of glycogen synthesis under food deprivation conditions. As the negative regulator, FoxO regulates glucose and lipid metabolism through the transcriptional activation of certain genes [FoxO was upregulated during fasting and downregulated after re-feeding. Furthermore, the expression of the IIS and ILPR-mediated signaling pathway genes was inhibited under low culture temperature, except for the negative regulator FoxO. These results confirm that the roles of the IIS and ILPR-mediated signaling pathways are limited, and that ILPR could transduce signaling from the active ternary complex “ILP–IGFBP–ALS” to the downstream PI3K-AKT and RAS-MAPK signaling pathways, regulating the energy metabolism and cell proliferation of C. gigas , and the phylogenetic trees were constructed using the neighbor-joining (NJ) approach in MEGA7 [The amino acid sequences of the IIS signaling pathway genes—including ILPR, IRS, IGFBP and IGFALS—from Homo sapiens, Gallus gallus, Xenopus laevis, Anolis carolinensis, Danio rerio, Limulus polyphemus, Musca domestica, Tetranychus urticae, Aplysia californica, Mizuhopecten yessoensis, Octopus bimaculoid, Pomacea canaliculata, Crassostrea virginica, and Crassostrea gigas were retrieved from the NCBI database. Detailed information on these sequences is provided in in MEGA7 . The relelongation factor 1α (EF1α), were used to assess primer efficiency, and the primer sets with an efficiency of 90–110% were used for real-time PCR analysis. The real-time PCR reactions were carried out in a LightCycler 480 machine  in a 20 μL system with a mixture of 10 μL 2×SYBR Premix ExTaq , 2.0 μL diluted cDNA or double-distilled water as a negative control, 6 μL PCR-grade water, and 1.0 μL of each primer (10 μM). The PCR reactions were initiated via denaturation at 95 °C for 3 min, followed by 40 amplification cycles at 95 °C for 15 s and at 60 °C for 30 s. Dissociation protocols were used to measure melting curves. The raw data of all the qRT-PCR were provided in t-test were used to determine statistical differences for single or multiple comparisons, respectively. The statistical significance was set as p < 0.05, as indicated by the asterisk symbol or by values with different letters.All of the primers used for real-time PCR were designed using Primer Express software , and are provided in ILPR, IRS, IGFBPRP, IGFALS, PI3K, PDK, AKT, RAS, MEK, ERK, PTEN, TOR, FoxO, GSK3β, and S6K—were determined in the adductor muscles of “Haida No.1” and wild oysters. Total RNA was extracted using Trizol Reagent  and reversely transcribed into cDNA according to the PrimeScript™ RT reagent Kit with gDNA Eraser  , according to the manufacturer’s instructions. Real-time PCR was carried out as described in The expression profiles of the major genes in the IIS, PI3K-AKT, and RAS-MAPK signaling pathways—including C. gigas were randomly divided into three groups and used for the fasting and re-feeding experiments. Six oysters collected before the experiment were used as controls (as indicated by C); the other oysters were starved for fourteen days and then re-fed with frozen Chlorella ad libitum for thirty-six hours. Samples were collected on days 1, 3, 5, 7, and 14  during the fasting process, and at 1, 3, 6, 12, 24, and 36 h  after re-feeding, with 6 oysters at each time point. Various tissues—including the labial palp, gill, mantle, digestive gland, hematocytes, heart, visceral ganglia, and adductor muscle—were rapidly excised, frozen in liquid nitrogen, and then stored at −80 °C until use. Total RNA extraction and cDNA synthesis were performed as described above. The cDNA from all eight tissues at each sampling point was pooled into one sample, with equal amounts per tissue, and used for real-time PCR analysis according to the method described in Ninety eight-month-old Oyster umbo larvae of similar sizes (2.5 ± 0.3 mm) were stocked into 3 75 L cylindrical polyethylene vessels  and reared at 5 ℃, 15 ℃, and 25 ℃, respectively. Light was supplied by fluorescent lamps with 12:12 Light: Dark photoperiods. For each culture temperature, 3 random replicates were allocated, the survival and feeding situation of the larvae was monitored daily for 10-day culture, the shell height of 30 randomly collected larvae was measured, and the whole larvae were collected and deposited into RNAstore  until used for RNA expression analyses. ILPR, one IRS, one IGFBPRP, and seven IGFALSs in C. gigas. Expression profiling of these genes in fast-growing oysters and in slow-growing wild oysters suggested their critical roles in growth regulation. Nutrient abundance and ambient temperature greatly affected the expression profiles of these genes. Furthermore, the major genes of the ILPR-mediated PI3K-AKT and RAS-MAPK signaling pathways were all expressed at higher levels in the fast-growing “Haida No.1” oysters, and were significantly suppressed under low-temperature conditions. These observations suggest that the IIS signaling pathway is involved in the growth regulation of C. gigas through regulating food intake, nutrition metabolism, and cell proliferation. This work provides valuable information for further investigation on growth regulation mechanisms in mollusks, as well as other invertebrates.In summary, we identified and characterized one