Machine Learning-Based Interpretable Modeling for Subjective Emotional Dynamics Sensing Using Facial EMG.

Understanding the association between subjective emotional experiences and physiological signals is of practical and theoretical significance. Previous psychophysiological studies have shown a linear relationship between dynamic emotional valence experiences and facial electromyography (EMG) activities. However, whether and how subjective emotional valence dynamics relate to facial EMG changes nonlinearly remains unknown. To investigate this issue, we re-analyzed the data of two previous studies that measured dynamic valence ratings and facial EMG of the corrugator supercilii and zygomatic major muscles from 50 participants who viewed emotional film clips. We employed multilinear regression analyses and two nonlinear machine learning (ML) models: random forest and long short-term memory. In cross-validation, these ML models outperformed linear regression in terms of the mean squared error and correlation coefficient. Interpretation of the random forest model using the SHapley Additive exPlanation tool revealed nonlinear and interactive associations between several EMG features and subjective valence dynamics. These findings suggest that nonlinear ML models can better fit the relationship between subjective emotional valence dynamics and facial EMG than conventional linear models and highlight a nonlinear and complex relationship. The findings encourage emotion sensing using facial EMG and offer insight into the subjective-physiological association.

The widespread action observation/execution matching system for facial expression processing.

Observing and understanding others' emotional facial expressions, possibly through motor synchronization, plays a primary role in face-to-face communication. To understand the underlying neural mechanisms, previous functional magnetic resonance imaging (fMRI) studies investigated brain regions that are involved in both the observation/execution of emotional facial expressions and found that the neocortical motor regions constituting the action observation/execution matching system or mirror neuron system were active. However, it remains unclear (1) whether other brain regions in the limbic, cerebellum, and brainstem regions could be also involved in the observation/execution matching system for processing facial expressions, and (2) if so, whether these regions could constitute a functional network. To investigate these issues, we performed fMRI while participants observed dynamic facial expressions of anger and happiness and while they executed facial muscle activity associated with angry and happy facial expressions. Conjunction analyses revealed that, in addition to neocortical regions (i.e., the right ventral premotor cortex and right supplementary motor area), bilateral amygdala, right basal ganglia, bilateral cerebellum, and right facial nerve nucleus were activated during both the observation/execution tasks. Group independent component analysis revealed that a functional network component involving the aforementioned regions were activated during both observation/execution tasks. The data suggest that the motor synchronization of emotional facial expressions involves a widespread observation/execution matching network encompassing the neocortex, limbic system, basal ganglia, cerebellum, and brainstem.

Regulation of electron transfer in the terminal step of the respiratory chain.

In mitochondria, electrons are transferred along a series of enzymes and electron carriers that are referred to as the respiratory chain, leading to the synthesis of cellular ATP. The series of the interprotein electron transfer (ET) reactions is terminated by the reduction in molecular oxygen at Complex IV, cytochrome c oxidase (CcO) that is coupled with the proton pumping from the matrix to the inner membrane space. Unlike the ET reactions from Complex I to Complex III, the ET reaction to CcO, mediated by cytochrome c (Cyt c), is quite specific in that it is irreversible with suppressed electron leakage, which characterizes the ET reactions in the respiratory chain and is thought to play a key role in the regulation of mitochondrial respiration. In this review, we summarize the recent findings regarding the molecular mechanism of the ET reaction from Cyt c to CcO in terms of specific interaction between two proteins, a molecular breakwater, and the effects of the conformational fluctuation on the ET reaction, conformational gating. Both of these are essential factors, not only in the ET reaction from Cyt c to CcO, but also in the interprotein ET reactions in general. We also discuss the significance of a supercomplex in the terminal ET reaction, which provides information on the regulatory factors of the ET reactions that are specific to the mitochondrial respiratory chain.

A short-term intervention of ingesting iron along with methionine and threonine leads to a higher hemoglobin level than that with iron alone in young healthy women: a randomized, double-blind, parallel-group, comparative study.

PURPOSE: Enhancing iron absorption and utilization is important for amelioration iron status faster and thereby, for improving quality of life. Dietary protein and amino acids, including methionine and threonine, have been reported to facilitate the absorption and utilization of dietary iron. Here, we investigated the effect of combined ingestion of methionine, threonine, and iron on the improvement of iron status during a short-term intervention, by comparing that with iron ingestion alone in healthy young women. METHODS: This was a randomized, double-blind, parallel-group, comparative study with 45 participants (aged 20-39) randomly assigned to three groups (n = 15 each): one group was administered 200 mg methionine, 400 mg threonine, and 6 mg iron once daily (FEMT); another ingested 6 mg iron alone (FE); and the third group ingested a placebo (PCG). Blood samples and dietary nutrient data were collected before the intervention (week 0) and after 2, 4, and 6 weeks. Serum iron, hemoglobin, transferrin, and ferritin levels were measured. RESULTS: Blood hemoglobin levels were significantly higher in the FEMT than in the FE group (P < 0.05) at week 4. Serum iron, transferrin, and ferritin levels were not changed across groups. In addition, our analyses showed that the observed increase in hemoglobin levels was affected by the intervention rather than changes in dietary nutrient intake. CONCLUSIONS: Ingestion of methionine and threonine with low doses of iron leads to a higher hemoglobin levels than that with iron alone in a short period of 4 weeks. TRIAL REGISTRATION: University Hospital Medical Information Network Clinical Trial Registry (UMIN000046621).

Crosstalk in Facial EMG and Its Reduction Using ICA.

There is ample evidence that electromyography (EMG) signals from the corrugator supercilii and zygomatic major muscles can provide valuable information for the assessment of subjective emotional experiences. Although previous research suggested that facial EMG data could be affected by crosstalk from adjacent facial muscles, it remains unproven whether such crosstalk occurs and, if so, how it can be reduced. To investigate this, we instructed participants (n = 29) to perform the facial actions of frowning, smiling, chewing, and speaking, in isolation and combination. During these actions, we measured facial EMG signals from the corrugator supercilii, zygomatic major, masseter, and suprahyoid muscles. We performed an independent component analysis (ICA) of the EMG data and removed crosstalk components. Speaking and chewing induced EMG activity in the masseter and suprahyoid muscles, as well as the zygomatic major muscle. The ICA-reconstructed EMG signals reduced the effects of speaking and chewing on zygomatic major activity, compared with the original signals. These data suggest that: (1) mouth actions could induce crosstalk in zygomatic major EMG signals, and (2) ICA can reduce the effects of such crosstalk.

Electromyographic Validation of Spontaneous Facial Mimicry Detection Using Automated Facial Action Coding.

Although electromyography (EMG) remains the standard, researchers have begun using automated facial action coding system (FACS) software to evaluate spontaneous facial mimicry despite the lack of evidence of its validity. Using the facial EMG of the zygomaticus major (ZM) as a standard, we confirmed the detection of spontaneous facial mimicry in action unit 12 (AU12, lip corner puller) via an automated FACS. Participants were alternately presented with real-time model performance and prerecorded videos of dynamic facial expressions, while simultaneous ZM signal and frontal facial videos were acquired. Facial videos were estimated for AU12 using FaceReader, Py-Feat, and OpenFace. The automated FACS is less sensitive and less accurate than facial EMG, but AU12 mimicking responses were significantly correlated with ZM responses. All three software programs detected enhanced facial mimicry by live performances. The AU12 time series showed a roughly 100 to 300 ms latency relative to the ZM. Our results suggested that while the automated FACS could not replace facial EMG in mimicry detection, it could serve a purpose for large effect sizes. Researchers should be cautious with the automated FACS outputs, especially when studying clinical populations. In addition, developers should consider the EMG validation of AU estimation as a benchmark.

Pleasant Stroke Touch on Human Back by a Human and a Robot.

Pleasant touching is an important aspect of social interactions that is widely used as a caregiving technique. To address the problems resulting from a lack of available human caregivers, previous research has attempted to develop robots that can perform this kind of pleasant touch. However, it remains unclear whether robots can provide such a pleasant touch in a manner similar to humans. To investigate this issue, we compared the effect of the speed of gentle strokes on the back between human and robot agents on the emotional responses of human participants (n = 28). A robot or a human stroked on the participants' back at two different speeds (i.e., 2.6 and 8.5 cm/s). The participants' subjective (valence and arousal ratings) and physiological (facial electromyography (EMG) recorded from the corrugator supercilii and zygomatic major muscles and skin conductance response) emotional reactions were measured. The subjective ratings demonstrated that the speed of 8.5 cm/s was more pleasant and arousing than the speed of 2.6 cm/s for both human and robot strokes. The corrugator supercilii EMG showed that the speed of 8.5 cm/s resulted in reduced activity in response to both human and robot strokes. These results demonstrate similar speed-dependent modulations of stroke on subjective and physiological positive emotional responses across human and robot agents and suggest that robots can provide a pleasant touch similar to that of humans.

Enhanced mirror neuron network activity and effective connectivity during live interaction among female subjects.

Facial expressions are indispensable in daily human communication. Previous neuroimaging studies investigating facial expression processing have presented pre-recorded stimuli and lacked live face-to-face interaction. Our paradigm alternated between presentations of real-time model performance and pre-recorded videos of dynamic facial expressions to participants. Simultaneous functional magnetic resonance imaging (fMRI) and facial electromyography activity recordings, as well as post-scan valence and arousal ratings were acquired from 44 female participants. Live facial expressions enhanced the subjective valence and arousal ratings as well as facial muscular responses. Live performances showed greater engagement of the right posterior superior temporal sulcus (pSTS), right inferior frontal gyrus (IFG), right amygdala and right fusiform gyrus, and modulated the effective connectivity within the right mirror neuron system (IFG, pSTS, and right inferior parietal lobule). A support vector machine algorithm could classify multivoxel activation patterns in brain regions involved in dynamic facial expression processing in the mentalizing networks (anterior and posterior cingulate cortex). These results indicate that live social interaction modulates the activity and connectivity of the right mirror neuron system and enhances spontaneous mimicry, further facilitating emotional contagion.

Precision Synthesis and Atomistic Analysis of Deep-Blue Cubic Quantum Dots Made via Self-Organization.

As a crystal approaches a few nanometers in size, atoms become nonequivalent, bonds vibrate, and quantum effects emerge. To study quantum dots (QDs) with structural control common in molecular science, we need atomic precision synthesis and analysis. We describe here the synthesis of lead bromide perovskite magic-sized nanoclusters via self-organization of a lead malate chelate complex and PbBr3- under ambient conditions. Millisecond and angstrom resolution electron microscopic analysis revealed the structure and the dynamic behavior of individual QDs─structurally uniform cubes made of 64 lead atoms, where eight malate molecules are located on the eight corners of the cubes, and oleylammonium cations lipophilize and stabilize the edges and faces. Lacking translational symmetry, the cube is to be viewed as a molecule rather than a nanocrystal. The QD exhibits quantitative photoluminescence and stable electroluminescence at ≈460 nm with a narrow half-maximum linewidth below 15 nm, reflecting minimum structural defects. This controlled synthesis and precise analysis demonstrate the potential of cinematic chemistry for the characterization of nanomaterials beyond the conventional limit.

Efp promotes growth of triple-negative breast cancer cells.

Triple-negative breast cancer (TNBC) is characterized by its high ability of invasiveness and metastasis, namely lacking expression of estrogen receptor (ER), progesterone receptor, and HER2. We previously demonstrated that estrogen responsive finger protein (Efp) plays a tumor-promotive role in ER-positive breast cancer, yet it remains to be addressed whether Efp contributes to TNBC pathophysiology. We here found that Efp mRNA and protein were abundantly expressed in TNBC patient-derived cells and MDA-MB-231 cells. Efp silencing significantly decreased the growth and migration of both TNBC cell models. Cell-cycle profiling showed a decrease in the S phase population upon Efp silencing. Moreover, exogenous Efp expression increased the growth, migration, and the percentages of S phase population of TNBC cells. Transcriptomic analysis in the Efp-silenced TNBC cells identified several candidate Efp targets including cell cycle-related genes CDCA7 and HELLS, whose contribution to cell growth were validated by siRNA-mediated gene silencing. These results suggest that Efp plays a tumor-promotive role in TNBC and can be a potential therapeutic target for the aggressive disease.

Rapid detection of neutral faces associated with emotional value.

Swift detection of faces with emotional meaning underlies fruitful social relationships. Although previous studies using a visual search paradigm have demonstrated rapid detection of emotional facial expressions, whether it is attributable to emotional/motivational significance remains to be clarified. We examined this issue by excluding the influence of visual factors on the rapid detection of faces with emotional meaning. First, participants were engaged in an associative learning task wherein neutral faces were associated with either monetary rewards, monetary punishments, or zero outcome in order for the neutral faces to acquire positive, negative, and no emotional value, respectively. Then, during the visual search task, the participants detected a target-neutral face associated with high reward or punishment from among newly presented neutral faces. In Experiment 1, neutral faces associated with high reward and punishment values were more rapidly detected than those without monetary outcomes. In Experiment 2, highly rewarded and highly punished neutral faces were more rapidly detected than neutral faces associated with low monetary reward/punishment. Analyses of ratings confirmed that the learned neutral faces acquired emotional value, and the reaction times were negatively related to arousal ratings. These results suggest that the emotional/motivational significance promotes the rapid detection of emotional faces.

Exploration of Emotion Dynamics Sensing Using Trapezius EMG and Fingertip Temperature.

Exploration of the physiological signals associated with subjective emotional dynamics has practical significance. Previous studies have reported that the dynamics of subjective emotional valence and arousal can be assessed using facial electromyography (EMG) and electrodermal activity (EDA), respectively. However, it remains unknown whether other methods can assess emotion dynamics. To investigate this, EMG of the trapezius muscle and fingertip temperature were tested. These measures, as well as facial EMG of the corrugator supercilii and zygomatic major muscles, EDA (skin conductance level) of the palm, and continuous ratings of subjective emotional valence and arousal, were recorded while participants (n = 30) viewed emotional film clips. Intra-individual subjective-physiological associations were assessed using correlation analysis and linear and polynomial regression models. Valence ratings were linearly associated with corrugator and zygomatic EMG; however, trapezius EMG was not related, linearly or curvilinearly. Arousal ratings were linearly associated with EDA and fingertip temperature but were not linearly or curvilinearly related with trapezius EMG. These data suggest that fingertip temperature can be used to assess the dynamics of subjective emotional arousal.

Triarylamine/Bithiophene Copolymer with Enhanced Quinoidal Character as Hole-Transporting Material for Perovskite Solar Cells.

Polytriarylamine is a popular hole-transporting materials (HTMs) despite its suboptimal conductivity and significant recombination at the interface in a solar cell setup. Having noted insufficient conjugation among the triarylamine units along the polymer backbone, we inserted a bithiophene unit between two triarylamine units through iron-catalyzed C-H/C-H coupling of a triarylamine/thiophene monomer so that two units conjugate effectively via four quinoidal rings when the molecule functions as HTM. The obtained triarylamine/bithiophene copolymer (TABT) used as HTM showed a high-performance in methylammonium lead iodide perovskite (MAPbI3 ) solar cells. Mesityl substituted TABT forms a uniform film, shows high hole-carrier mobility, and has an ionization potential (IP=5.40 eV) matching that of MAPbI3 . We fabricated a solar cell device with a power conversion efficiency of 21.3 % and an open-circuit voltage of 1.15 V, which exceeds the performance of devices using reference standard such as poly[bis(4-phenyl)(2,4,6-trimethylphenyl)amine (PTAA) and Spiro-OMeTAD.

Iron-Catalyzed Tandem Cyclization of Diarylacetylene to a Strained 1,4-Dihydropentalene Framework for Narrow-Band-Gap Materials.

Carbon bridging in a form of a strained 1,4-dihydropentalene framework is an effective strategy for flattening and stabilizing oligophenylenevinylene systems for the development of optoelectronic materials. However, efficient and flexible methods for making such a strained ring system are lacking. We report herein a mild and versatile synthetic access to the 1,4-dihydropentalene framework enabled by iron-catalyzed single-pot tandem cyclization of a diarylacetylene using FeCl2 and PPh3 as catalyst, magnesium/LiCl as a reductant, and 1,2-dichloropropane as a mild oxidant. The new annulation method features two iron-catalyzed transformations used in tandem, a reductive acetylenic carboferration and an oxidation-induced ring contraction of a ferracycle under mild oxidative conditions. The new method provides access not only to a variety of substituted indeno[2,1-a]indenes but also to their thiophene congeners, 4,9-dihydrobenzo[4,5]pentaleno[1,2-b]thiophene (CPTV) and 4,8-dihydropentaleno[1,2-b:4,5-b']dithiophenes (CTV). With its high highest occupied molecular orbital level and narrow optical gap, CTV serves as a donor unit in a narrow-band-gap non-fullerene acceptor, which shows absorption extending over 1000 nm in the film state, and has found use in a near-infrared photodetector device that exhibited an external quantum efficiency of 72.4% at 940 nm.

Transcriptomic analysis of hormone-sensitive patient-derived endometrial cancer spheroid culture defines Efp as a proliferation modulator.

Estrogen-responsive endometrial cancer (EC) is prevalent in uterine cancer. Its precise molecular mechanisms remain to be elucidated partly because of limited availability of estrogen-sensitive EC models recapitulating clinical pathophysiology. We previously established EC patient-derived cancer cell (EC-PDC) spheroid culture with high expression of estrogen receptor α (ERα). Using this EC-PDC, we study the transcriptional regulation and function of estrogen-responsive finger protein (Efp), a prototypic tripartite motif (TRIM) protein that modulates protein degradation and RNA processing. Intense estrogen-dependent EFP mRNA induction and high ERα occupancy to EFP estrogen responsive element (ERE) were observed in EC-PDC. Luciferase reporter gene assay showed that the ERE facilitates EFP transcriptional activity estrogen-dependently. siRNA-mediated Efp silencing in EC-PDC resulted in suppressed spheroid proliferation and altered gene expression profile, featuring downregulation of genes related to cell cycle (e.g., CDK6) and inflammation/immune responses (e.g., IL10RA, IL26, and IL6ST) while unaffected expression of cancer stemness-related markers. Taken together, EC-PDC spheroid culture is a powerful EC tool that enables to dissect Efp-mediated ERα signaling pathways as an estrogen-sensitive EC model. This study provides an insight into alternative EC therapeutic strategies targeting ERα-Efp axis.

Effects of localisation of uterine adenomyosis on outcome of in vitro fertilisation/intracytoplasmic sperm injection fresh and frozen-thawed embryo transfer cycles: a multicentre retrospective cohort study.

BACKGROUND: Uterine adenomyosis is a benign disease, common among women in their 40 and 50 s, characterised by ectopic endometrial tissue in the uterine myometrial layer. Adenomyosis causes infertility and has a negative effect on the outcomes of in vitro fertilisation (IVF)/intracytoplasmic sperm injection (ICSI) embryo transfer (ET) cycles. It has also been reported to have different characteristics depending on the adenomyotic lesion localisation. The effect of its localisation on IVF/ICSI-ET outcomes is unclear. This study aimed to investigate whether adenomyotic lesion localisation, assessed using magnetic resonance imaging (MRI), was associated with outcomes of IVF/ICSI-ET cycles. METHODS: This multicentre, joint, retrospective cohort study analysed the medical records of 67 infertile patients with adenomyosis who underwent IVF/ICSI with fresh and frozen-thawed ET at five participating facilities from January 2012 to December 2016 and for whom MRI data were available. Fifteen patients were excluded; therefore, the MRI data of 52 patients were evaluated by two radiologists. We assessed the localisation of and classified adenomyotic lesions into advanced (invades the full thickness of the uterine myometrium), extrinsic (localised on the serosal side), and intrinsic (localised on the endometrial side) subtypes. RESULTS: There were 40 advanced, nine extrinsic, and three intrinsic cases, and the outcomes of 100, 27, and nine ET cycles, respectively, were analysed. Pregnancy loss/clinical pregnancy and live birth rates of the advanced, extrinsic, and intrinsic groups were 64 % (16/25) and 9 % (9/100), 33.3 % (3/9) and 22.2 % (6/27), and 50 % (1/2) and 11.1 % (1/9), respectively. A logistic regression analysis adjusted for age, prior miscarriage, and body mass index showed that the extrinsic group had fewer pregnancy losses (odds ratio 0.06; 95 % confidence interval [CI]: 0.00-0.54, p = 0.026) and more live births (odds ratio 6.05; 95 % CI: 1.41-29.65, p = 0.018) than the advanced group. CONCLUSIONS: Adenomyotic lesions exert different effects on IVF/ICSI-ET outcomes. Thus, MRI assessments of adenomyosis in infertile patients are beneficial. Establishment of treatment plans based on adenomyotic lesion localisation should be considered.

Osmotic pressure effects identify dehydration upon cytochrome c-cytochrome c oxidase complex formation contributing to a specific electron pathway formation.

In the electron transfer (ET) reaction from cytochrome c (Cyt c) to cytochrome c oxidase (CcO), we determined the number and sites of the hydration water released from the protein surface upon the formation of the ET complex by evaluating the osmotic pressure dependence of kinetics for the ET from Cyt c to CcO. We identified that ∼20 water molecules were dehydrated in complex formation under turnover conditions, and systematic Cyt c mutations in the interaction site for CcO revealed that nearly half of the released hydration water during the complexation were located around Ile81, one of the hydrophobic amino acid residues near the exposed heme periphery of Cyt c. Such a dehydration dominantly compensates for the entropy decrease due to the association of Cyt c with CcO, resulting in the entropy-driven ET reaction. The energetic analysis of the interprotein interactions in the ET complex predicted by the docking simulation suggested the formation of hydrophobic interaction sites surrounding the exposed heme periphery of Cyt c in the Cyt c-CcO interface (a 'molecular breakwater'). Such sites would contribute to the formation of the hydrophobic ET pathway from Cyt c to CcO by blocking water access from the bulk water phase.

Assessing Automated Facial Action Unit Detection Systems for Analyzing Cross-Domain Facial Expression Databases.

In the field of affective computing, achieving accurate automatic detection of facial movements is an important issue, and great progress has already been made. However, a systematic evaluation of systems that now have access to the dynamic facial database remains an unmet need. This study compared the performance of three systems (FaceReader, OpenFace, AFARtoolbox) that detect each facial movement corresponding to an action unit (AU) derived from the Facial Action Coding System. All machines could detect the presence of AUs from the dynamic facial database at a level above chance. Moreover, OpenFace and AFAR provided higher area under the receiver operating characteristic curve values compared to FaceReader. In addition, several confusion biases of facial components (e.g., AU12 and AU14) were observed to be related to each automated AU detection system and the static mode was superior to dynamic mode for analyzing the posed facial database. These findings demonstrate the features of prediction patterns for each system and provide guidance for research on facial expressions.

Consistency between chromosomal status analysis of biopsied human blastocyst trophectoderm cells and whole blastocyst cells.

PURPOSE: This study investigated the consistency between results of preimplantation genetic testing for aneuploidy performed on trophectoderm (TE) cells and remaining blastocyst cells. METHODS: TE biopsy was performed on 29 surplus cryopreserved human blastocysts. Biopsy samples and remaining blastocysts were processed using the VeriSeq PGS kit, and chromosomal statuses were compared by next-generation sequencing. RESULTS: Discordance was observed in the chromosomal status of 11 out of 29 blastocysts between the biopsied TE and remaining blastocysts. Concordance was observed in 11 of 12 blastocysts classified as euploid by TE biopsy and in 7 of 17 blastocysts classified as aneuploid. There was 100% concordance (7/7) in cases diagnosed as aneuploid with no mosaicism by TE biopsy. However, discordance was observed in all 10 cases showing mosaicism or partial chromosomal abnormality. CONCLUSION: Chromosomal status analysis based on TE biopsy does not accurately reflect the chromosomal status of the whole blastocyst. The chromosomal status is usually the same between the TE and remaining blastocyst cells in cases diagnosed as euploid or aneuploid with no mosaicism. However, mosaic blastocysts and those with other types of structural rearrangements have a higher risk of inconsistency, warranting caution during embryo selection.

PDZK1-interacting protein 1 (PDZK1IP1) traps Smad4 protein and suppresses transforming growth factor-ß (TGF-ß) signaling.

Transforming growth factor (TGF)-ß signaling in humans is stringently regulated to prevent excessive TGF-ß signaling. In tumors, TGF-ß signaling can both negatively and positively regulate tumorigenesis dependent on tumor type, but the reason for these opposite effects is unclear. TGF-ß signaling is mainly mediated via the Smad-dependent pathway, and herein we found that PDZK1-interacting protein 1 (PDZK1IP1) interacts with Smad4. PDZK1IP1 inhibited both the TGF-ß and the bone morphogenetic protein (BMP) pathways without affecting receptor-regulated Smad (R-Smad) phosphorylation. Rather than targeting R-Smad phosphorylation, PDZK1IP1 could interfere with TGF-ß- and BMP-induced R-Smad/Smad4 complex formation. Of note, PDZK1IP1 retained Smad4 in the cytoplasm of TGF-ß-stimulated cells. To pinpoint PDZK1IP1's functional domain, we created several PDZK1IP1 variants and found that its middle region, from Phe40 to Ala49, plays a key role in its Smad4-regulating activity. PDZK1IP1 knockdown enhanced the expression of the TGF-ß target genes Smad7 and prostate transmembrane protein androgen-induced (TMEPAI) upon TGF-ß stimulation. In contrast, PDZK1IP1 overexpression suppressed TGF-ß-induced reporter activities, cell migration, and cell growth inhibition. In a xenograft tumor model in which TGF-ß was previously shown to elicit tumor-promoting effects, PDZK1IP1 gain of function decreased tumor size and increased survival rates. Taken together, these findings indicate that PDZK1IP1 interacts with Smad4 and thereby suppresses the TGF-ß signaling pathway.