Quality staff-resident communication is crucial to promote outcomes in nursing home residents with dementia requiring assistance during mealtimes. Better understanding of staff-resident language characteristics in mealtime interactions help promote effective communication, yet evidence is limited. This study aimed to examine factors associated with language characteristics in staff-resident mealtime interactions.This was a secondary analysis of 160 mealtime videos involving 36 nursing staff and 27 residents with moderately severe to severe dementia in 9 nursing homes. Mixed-effects models was used to examine the relationships between factors and language characteristics in staff-resident mealtime interactions. The independent variables were speaker (resident vs. staff), utterance quality (negative vs. positive), intervention (pre- vs. post-communication intervention), and resident dementia stage and comorbidities. The dependent variables were expression length (number of words in each utterance) and addressing partner by name (whether staff or resident named their partner in each utterance). All models included staff, resident, and staff-resident dyad as random effects.Staff  predominated conversations and had more positive, longer utterances than residents . As residents progressed from moderately severe to severe dementia, both residents and staff produced shorter utterances . Staff (18%) named residents more often than residents  and when assisting residents with more severe dementia .Staff-resident communication was primarily positive, staff-initiated, and resident-oriented. Utterance quality and dementia stage were associated with staff-resident language characteristics. Staff play a critical role in mealtime care communication and should continue to initiate resident-oriented interactions using simple, short expressions to accommodate resident declining language abilities, particularly those with severe dementia. Staff should practice addressing residents by their names more frequently to promote individualized, targeted, person-centered mealtime care. Future work may further examine staff-resident language characteristics at other levels of language using more diverse samples. People with dementia in residential care settings  often exhibit cognitive, functional, and behavioral challenges during mealtime such as disorientation to meal time/place, swallowing difficulties, and resistance to food/care, and require varied levels of support from staff . MealtimWhile mealtime can be challenging for people with dementia, it is a good opportunity to establish and practice pperson-centered interactions and communication. While interacting and communicating with one another, participants reciprocally make an informational exchange through verbal and nonverbal expressions between staff and residents, which can be an important component of person-centered care especially in residential care settings –5. PersoMultiple factors at the resident-, staff-, and context-levels are associated with person-centered communication between nursing staff and residents with dementia during mealtime. Resident-level factors include social demographics , comorbidities, function , behaviors , verbal communication skills , and values  . AdditioMealtime involves dynamic, interactive, and complex interactions among staff-resident dyads. Among the identified multi-level factors associated with person-centered communication in mealtimes, the majority are related to nursing staff and can be modified, illustrating staff’s critical role as active contributors in influencing the quality of communication and addressing the needs and preferences of residents with dementia . NursingVerbal communication plays a key role in interacting with one another and achieving desirable outcomes for staff and residents during mealtimes. On that account, understanding the characteristics of language produced during mealtimes is crucial to optimize mealtime interactions as well as behaviors, function, nutrition, and quality of life for residents with dementia. Analysis of language can be at different levels, including word , syntax/sentence (syntactic complexity of sentences), speech (speech rate or intelligibility), and discourse (coherence and cohesion of conversations) . While pThis study aimed to (1) describe word-level language characteristics  in staff and resident utterances during mealtime interactions, (2) examine the associations between staff-level factors  and language characteristics, and (3) examine the associations between resident-level factors  and language characteristics.This study was a secondary behavioral analysis of videotaped mealtime observations collected from a randomized clinical trial during 2011–2014. The parent trial evaluated the effect of the Changing Talk (CHAT) intervention on staff elderspeak communication and resident resistiveness to care . ElderspIn the parent trial, a convenience sample of 127 nursing staff and 83 residents were recruited from 13 NHs in Kansas, USA. NHs located within two hours from the research site and providing care for residents with dementia were recruited. In each NH, residents were eligible if they had (1) a dementia diagnosis of any type or stage based on medical records, (2) staff-reported resistiveness to care, (3) long-stay status, (4) hearing capacity based on medical records , and (5) a surrogate decision maker to provide informed consent were eligible . Staff wVideos were recorded to capture staff-resident communication during daily care activities, including mealtimes, and were archived in the parent trial. Video recordings were collected on two days at each time point in the parent study. Prior to the first recording session, a practice recording session was completed by a trained research assistant (videographer) using a mini handheld digital video recorder to minimize the Hawthorne effect by (1) allowing residents to adjust to the novelty of being recorded, (2) allowing the videographer to become familiar with daily care routines, (3) evaluating any adverse impact on residents, and (4) establishing behaviors indicating that recording should be discontinued. Recording did not take place during the practice recording session, but the videographer observed and made notes of resident reactions to the presence of the camera and the videographer. After the practice recording session, it was expected that residents would be familiar with videographers’ faces and existence and behave more naturally when formal recordings occurred. Practice recording was not repeated at the remaining data collection points.Video recordings were collected on day and evening shifts of weekdays. Recordings primarily captured morning care when staff-resident care interactions are most intensive, except when resident resistiveness to care typically occurred at another time of the day based on staff report. The time of day and specific activity recorded were kept constant as much as possible for each resident. Recordings occurred in resident rooms as well as public areas. To facilitate resident de-sensitization to the recording situation, the camera was placed prior to the start of the care activity. Before each recording session, the videographer reminded the resident and staff that recording would take place and obtained oral consent/assent of video recording from the resident and staff. Those recordings that incidentally included persons who did not consent to be in the study were deleted. The videographer stood in an unobtrusive location while the recording occurred, and reviewed video footage on site to ensure the quality of recording. When a resident was recorded during a care activity, including mealtime, the recording started when communication between staff and resident started or continued, and ended when the dyadic communication discontinued or stopped. Therefore, segments of mealtime rather than full mealtime were recorded in the parent study .The archived videos from the parent study were screened for this study. Videos were eligible if they: (1) lasted for ≥ 1 min, (2) captured mealtime interactions between one resident and one staff, and (3) captured utterances with adequate audio quality. A total of 1,748 videos were screened, from which, 1,588 videos were excluded due to lasting < 1 min (n = 63), capturing other activities rather than mealtimes , involving more than one staff and/or more than one resident (n = 34), and poor audio quality (n = 5). Thus, 160 videos were eligible for this study, of which, 110 were collected prior to the staff communication training (pre-intervention) and 50 were after the training (post-intervention) .In this study, staff and resident utterances were transcribed and coded using the refined Cue Utilization and Engagement in Dementia (CUED) Mealtime Video-Coding Scheme in 2019 using Noldus Observer® 14.0  . In the Four coders were trained by the first author following a standard training and coding manual. After establishing inter-coder reliability, each of the four trained coders coded a subset of videos independently. Each utterance (a statement or question) was assigned a code. All utterances were coded as point events, where onset time (vs. offset time) of utterance was coded. Detailed coding process and conceptual and operational definitions of all codes are described elsewhere , 23, 24.For this study, two dependent variables—expression length and addressing partner by their name— representing word-level language characteristics were coded for each transcribed utterance of residents and staff.Expression length, a continuous variable operationalized as the number of words produced in each utterance.Addressing the partner by their name, a binary variable operationalized as whether staff or resident named their dyadic partner  in each utterance.For this study, the independent variables included speaker, utterance quality, and intervention, in addition to resident dementia stage and comorbidities.Speaker, a binary variable operationalized as whether resident or staff produced each utterance.Utterance quality, a binary variable operationalized as whether each utterance was coded as positive vs. negative in quality based on the refined CUED.Intervention, a binary variable operationalized as whether videos were collected before or after the dementia communication training was delivered to staff (pre- vs. post-CHAT intervention).Characteristics of facilities, staff, residents, and utterances were described using descriptive statistics . Poisson-link mixed-effects model was used to fit expression length with speaker (resident vs. staff), utterance quality (negative vs. positive), and intervention (pre- vs. post-intervention) as fixed effects. Poisson distribution is appropriate as the dependent variable, expression length, is a count of the number of occurrences during a defined time interval . Logit-ls were obtained through Institutional Review Boards of universities where the studies were conducted. In the parent study, NHs were first enrolled and randomized to the intervention or waiting list control group. In each enrolled NH, staff and resident participants were provided with information about the study and recruited with written consent (staff) and surrogate consent from the resident’s Legally Authorized Representative and resident assent (resident) .This study as a secondary data analysis of the videos collected from the parent study was approved by Institutional Review Board . A data use agreement between the institution of parent study PI and the institution of the secondary data analysis study PI (first author of this manuscript) was established (see Supplementary file #2), indicating the approved use of videos from residents and staff who indicated consent in the use of their video recordings in future research studies in their written consent forms. All videos used in the study were stored in the Research Data Storage Service (RDSS) at the University of Iowa, a secured system for backups, archiving, and storing research data files in the institution system.The 160 videos lasted between 1 and 23.8 min . Most of the videos were recorded in the dining rooms  and the rest were recorded in the residents’ own room . In three quarters of the videos , residents were seated in a chair, and in the other 25% of the videos, residents were seated in a wheelchair. The 160 videos that were used in this study involved 27 residents and 36 nursing staff (53 unique staff-resident dyads) in 9 NHs. In this study, while each resident was assisted by one staff in each videotaped mealtime interaction, the resident could be assisted by the same or different staff across all the videorecorded mealtime interactions, resulting in one or multiple unique staff-resident dyads. Among the 27 residents, 9 residents, respectively, were assisted by 1 staff in the videos that captured the residents, resulting in 9 unique staff-resident dyads; 12 residents, respectively, were assisted by 2 different staff in the videos that captured the residents, resulting in 24 unique staff-resident dyads; 4 residents, respectively, were assisted by 3 different staff in the videos that captured the residents, resulting in 12 unique staff-resident dyads; 2 residents, respectively, were assisted by 4 different staff in the videos that captured the residents, resulting in 8 unique staff-resident dyads.The nine NHs ranged from 43 to 163 beds in size (Median = 60 beds) and were distributed evenly on location , profit status , and quality ratings . Five NHs had one or more memory care units.Resident characteristics were collected through medical records. As show in Table Staff characteristics were collected using self-report surveys. As shown in Table Staff (utterances n = 2990) spoke three times more often than residents  and utterance quality  and utterance quality , and the three-way interaction between speaker, utterance quality, and intervention  remained significant. Expression length was significantly associated with resident dementia stage , suggesting that as resident dementia stage progresses from moderately severe into severe, both residents and staff are likely to produce shorter utterances. Expression length was not significantly associated with resident comorbidities .The model revealed addressing the partner by their name was significantly associated with speaker , indicating staff addressed residents by their name more often than residents addressing staff by their name during mealtimes Table . UtteranThis study described staff-resident language characteristics  during mealtime interactions and examined their associations with speaker, utterance quality, staff reception of dementia communication intervention, and resident characteristics . While staff predominated dyadic conversations, residents were also involved in the conversations. While both staff and residents predominately used positive communication, staff used more positive communication comparatively. Not surprisingly, staff spoke longer sentences and named their partner  more often than residents, indicating staff-resident mealtime interactions were primarily staff-initiated and resident-oriented. Most of the mealtime interactions in this study occurred in the dining areas (vs. resident own room), representing common mealtime care practices in NHs. Residents were exposed to different environmental stimuli including food, conversations, and assistance provided by staff as well as occupied with eating food, and therefore, were less dominant in dyadic interactions.Interestingly, staff produced longer utterances in their positive (vs. negative) utterances while residents produced longer utterances in their negative (vs. positive) utterances; however, the difference between positive and negative utterances in both staff (0.87 words) and residents (-0.84 words) was small. Such findings may be due to the unbalanced distribution of positive vs. negative utterances in staff and residents. Utterance quality in the study sample had limited variations . Further analysis indicated that staff positive utterances n = 2963) were distributed across all eight codes see  section 63 were dThe study showed that staff and resident expression length was associated with resident dementia stage. This is consistent with prior work that residents with neurodegenerative deterioration produce more frequent, easier words and shorter, simpler sentences , 12. ResThe study showed that staff expressions became shorter and simpler when providing mealtime care to residents at the stage of severe (vs. moderately severe) dementia, possibly because staff adapted their communication to accommodate residents’ decline in understanding and mastery of language due to residents’ cognitive decline. Staff as cognitively intact individuals were able to adjust expressions based on their partners’ needs in social interactions, such as using shorter, simpler phrases in communicating with residents with dementia . The stuStaff named residents more often as resident progresses from moderately severe to severe dementia stage. Addressing residents by their name during dyadic communication is a critical strategy of person-centered care to acknowledge resident identity, show respect, and establish emotional/personal connection, as well as to engage residents in activities . While aStaff intervention to avoid elderspeak was not associated with staff-resident language characteristics in this study, possibly because the dementia communication training tested in the parent trial focused on reducing elderspeak  by staff when communicating with residents during care activities in general, not necessarily focusing on other communication approaches or activities during mealtimes . A similThe sample in the parent clinical trial focused on residents with staff-reported resistiveness to care during daily activities, which is a population that may require additional attention in dyadic communication. Resident resistive behaviors are considered a way to communicate their needs, preferences, and wants and maybe the only way of communication for residents who cannot verbalize or have lost their language ability. Recent work showed that the use of person-centered verbal cues was associated with increased food intake among residents who were compliant, and were associated with decreased intake among those with resistive behaviors during mealtimes . ResidenWhile current mealtime care practice in NHs may evolve due to various factors, such as the shift of care from task-oriented to person-oriented, the COVID-19 pandemic and its following consequences, and the understaffing, the video sample collected between 2011 and 2014 reflects current dyadic mealtime care interactions to certain degrees for three reasons. First, the videos reflect the nature of the complex and dynamic staff-resident mealtime verbal interactions that feature different levels of recognition, negotiation, validation, and facilitation. Second, the CUED tool used to assess staff and resident utterance during mealtime interactions was developed based on extensive systematic reviews of literature on dementia mealtime care research including person-centered and task-centered care measures , 41, 42.The use of videotaped observations and a validated behavioral tool in this study captured the complexity and interactivity of dyadic mealtime interactions. The use of videos allows for repetitive reviews of dyadic verbal interactions for precise and accurate coding and in-depth analysis of utterance and language characteristics. The CUED is developed based on extensive literature review of dementia mealtime care research and captures a comprehensive list of staff and resident verbal behaviors from both positive and negative perspectives, addressing the current gaps in characterizing dyadic mealtime verbal communication.The video sample captured primarily segments of meals  and 1:1  interactions. Future research may examine staff-resident language characteristics using full-meal observations that capture varied dynamic complexity of mealtime interactions. Further, this study primarily focused on the analysis of word-level language characteristics of staff and residents because utterances of nursing staff and residents with dementia in this study were short and simple, and conversations jumped from one topic to another due to the nature of dynamic and fluid dyadic mealtime interactions, which makes analysis at other language levels  challenging. Future work may expand the analysis to other levels of language  using more diverse samples.While videos were mostly collected during morning care in the parent study, the time of the meal were not recorded, and therefore the role of different types of meals  could not be examined. Videotaped observations collected from pre- and post-intervention were used in this study. While measures  have been taken to minimize the Hawthorne effect of videotaping on staff-resident interactions, staff may perform differently from usual care due to social desirability. The video sample may not reflect the most current mealtime care practice and future work can validate the findings using up-to-date mealtime observations.Lastly, staff participants were primarily nursing care providers and resident participants had exclusively moderately-severe to severe dementia with staff-reported resistiveness to care in NHs. Therefore, findings have limited generalizability to other staff-resident populations in other care settings.Quality dyadic communication is crucial to promote care quality as well as resident behaviors, function, hydration, and nutrition during mealtimes. This study provided preliminary evidence on the associations of staff-resident language characteristics with utterance quality and resident dementia stage. Residents with dementia have limited language ability which influences the complexity and length of conversations they can initiate and understand. Our findings indicate there is room to increase the use of person-centered care strategies including simple, short expressions and addressing residents by name during mealtime care practice of people with dementia. Future work needs to examine staff-resident language characteristics at the word and other levels of language using larger, diverse samples such as full-meal observations.


The immune and endocrine dysfunctions of white adipose tissue are a hallmark of metabolic disorders such as obesity and type 2 diabetes. In humans, white adipose tissue comprises distinct depots broadly distributed under the skin (hypodermis) and as internal depots . Depot-specific ASCs could account for visceral and subcutaneous adipose tissue properties, by regulating adipogenesis and immunomodulation. More importantly, visceral and subcutaneous depots account for distinct contributions to obesity and its metabolic comorbidities. Recently, distinct ASCs subpopulations were also described in subcutaneous adipose tissue. Interestingly, the superficial layer closer to the dermis shows hyperplastic and angiogenic capacities, whereas the deep layer is considered as having inflammatory properties similar to visceral. The aim of this focus review is to bring the light of recent discoveries into white adipose tissue heterogeneity together with the biology of distinct ASCs subpopulations and to explore adipose tissue 3D models revealing their advantages, disadvantages, and contributions to elucidate the role of ASCs in obesity development. Recent advances in adipose tissue organoids opened an avenue of possibilities to recreate the main cellular and molecular events of obesity leading to a deep understanding of this inflammatory disease besides contributing to drug discovery. Furthermore, 3D organ-on-a-chip will add reproducibility to these adipose tissue models contributing to their translation to the pharmaceutical industry. The immune and endocrine dysfunctions of white adipose tissue are a hallmark of metabolic disorders such as obesity and type 2 diabetes. These dysfunctions negatively impact on insulin sensitivity, metabolism, and promote local and systemic inflammation ,2,3. In Subcutaneous adipose tissue can be divided into two main layers—superficial and deep—delimited by a conjunctive fascia. Recently, distinct ASCs subpopulations were also described in subcutaneous adipose tissue. Interestingly, the superficial layer closer to the dermis shows hyperplastic and angiogenic capacities, whereas the deep layer is considered as having inflammatory properties similar to visceral. These ASCs subpopulations have distinct contributions to subcutaneous adipose tissue physiology and may represent the key to the recovery of ASC properties in an inflammatory scenario .Currently, the inability of traditional monolayer culture and animal models to reproduce molecular and cellular events of human tissues and organs is a consensus in the scientific literature. Three-dimensional cell culture models such as spheroids and organoids are increasingly used as models more faithful to the human cell physiology, creating a need for the development of functional adipose tissue 3D models .The aim of this focus review is to bring the light of recent discoveries into white adipose tissue heterogeneity together with the biology of distinct ASCs subpopulations and to explore adipose tissue 3D models revealing their advantages, disadvantages, and contributions to elucidate the role of ASCs in obesity development. We further provide considerations on which methodological and technological advances may be necessary to improve obese adipose tissue in vitro modeling. This review may guide further studies in which both ASCs from subcutaneous and visceral depots and from ASCs subpopulations of distinct layers of subcutaneous adipose tissue may unravel cellular and molecular mechanisms as potential targets for the recovery of ASC and adipose tissue function in the context of obesity.Knowledge of white adipose tissue physiology and metabolic dysfunctions has expanded significantly in recent decades. Once regarded as a simple inert energy storage tissue, white adipose tissue is now recognized as a biologically dynamic organ. More importantly, white adipose tissue has diffuse locations throughout the body as fat depots. Different physiologies, cell compositions, and functions are characterized for visceral and subcutaneous depots, and recently, for the different layers of the subcutaneous.White adipose tissue (WAT) acts primarily as a regulatory center for the homeostasis of the body’s energy metabolism. This is accomplished through the regulation of adipocyte lipid storage or release in response to the body’s energy demands, regulation of blood glucose levels due to the high insulin sensitivity of adipocytes , and thrWAT is one of the body’s pathways that, through remodeling of adipose tissue, allows adaptation to metabolic challenges posed by different external environmental changes, including food deficit, energy excess, stress, infection, or cold . CrosstaWAT depots with distinct expansion and remodeling patterns are widely distributed in the human body, comprising mainly subcutaneous and visceral depots. They have intrinsic biological differences  and diff+), termed “fibro-inflammatory progenitors” , regulates visceral WAT macrophage accumulation in mice fed a high-fat diet in a Toll-like receptor 4-dependent manner resulting in WAT dysfunction [We have previously shown that the SVF of obese abdominal SAT in humans has the highest content of preadipocytes compared to the VAT. More importantly, adipocyte stem and progenitor cells’ in vitro counterparts, here referred to as the adipose-derived stromal/stem cells (ASC), are depot-specific, with an inverse relationship between the adipogenic and immunogenic status: ASC from VAT are more pro-inflammatory in terms of cytokine secretion, with lower adipogenic potential . We havefunction . PDGFRβ+function . These aRecent efforts on single-cell level profiling of adipocyte progenitors and on clonal analysis of their in vitro counterparts (ASC) derived from different WAT depots of donors with a variety of metabolic phenotypes have clarified the understanding of ASC heterogeneity, their depot-dependent characteristics, and their contribution to the obesity scenario . Vijay aBST2) was identified as a marker of visceral ASC in a non-obese state in humans. However, the difference in BST2 expression between subcutaneous and visceral ASC is more pronounced in obesity and with insulin resistance. In addition, in vitro-derived adipocytes from non-obese ASCs of VAT have lower gene expression of adipogenic markers and higher gene expression of immunogenic markers than those derived from SAT [−CD45−CD29+ SVF cells. Interestingly, adipocytes derived from each progenitor subtype display distinct metabolic and endocrine phenotypes. Furthermore, adipocyte progenitors were more frequent in gluteo-femoral SAT than in abdominal SAT and VAT, and the frequency of adipocyte progenitor subtypes varies among donors with type 2 diabetes. ASCs showing depot-specific genetic [The intrinsic differences in ASC derived from VAT and SAT are also present in a non-obese state and are retained during obesity. The immunogenic factor bone marrow stromal cell antigen 2  which allows the dermis to be a support tissue for the skin. There is an ECM signature of the different fibroblasts subpopulation . The papIn addition to their role in ECM regulation, fibroblasts also interact with other cell types such as ASCs, located in SAT just beneath the reticular dermis. Haydont and collaborators  recentlyIf fibroblasts can interact with ASCs modulating their behavior, ASCs can also do the same with fibroblasts, as suggested by in vitro assays.ASC’s conditioned culture medium increased proliferation of dermal fibroblasts ,66. ThisThe beneficial effect of ASCs in dermis is even observed in altered conditions, such as keloid scar, since TGF-beta1-induced myofibroblast differentiation and human dermal fibroblasts’ function were inhibited by ASC’s conditioned culture medium . In addiTo conclude, the scientific literature supports the crosstalk between dermal fibroblasts and ASCs that impact skin quality. Soluble factors secreted by ASCs seem to regulate the skin microenvironment according to the needs. Surprisingly, a recent study showed that diabetic microenvironment-preconditioned ASCs effectively strengthen the capacity against inflammation and modulate the progress of long-term T2D complications . In addiASCs are capable of reproducing in vitro their corresponding adipose tissue origin. However, 2D culture has severe limitations. Long-term cell culture is not possible using monolayers, limiting cell differentiation and disease models. More importantly, the adipose tissue microenvironment is complex showing cross-talk among adipocytes, pre-adipocytes, ASCs, endothelial and immune cells. The first strategy for adipose tissue 3D models had a focus on soft tissue reconstruction in the plastic surgery field. The understanding of the adipose tissue as an endocrine organ and obesity as an inflammatory chronic disease boosted these 3D models. Currently, the main concern is to recapitulate physiological functions of adipose tissue and its alterations during obesity development with corresponding exogenous stimulus and cellular components.In 1993, Langer and Vacanti  establisDespite the success of the scaffold-based strategies for soft tissue reconstruction in animal models ,92, thisBy definition, a spheroid is a spheroidal 3D structure constituted by mature, progenitor, or stem cells and extracellular matrix molecules. On the other hand, organoids must be formed from stem cells derived from healthy or pathological tissues, having at least one physiological function corresponding to the desired organ and not necessarily showing a spheroidal 3D structure . HoweverDue to the chronic inflammatory scenario established during obesity, obese adipose tissue in vitro 3D models intend to mimic this inflammatory microenvironment using different approaches. Turner and collaborators  showed tFatty acids can also act as a proinflammatory stimulus. Recently, Pieters and collaborators (2022) treated human ASC embedded into hydrogel with fatty acids after adipogenic differentiation. The resulting adipocytes showed obese characteristics such as hypertrophy, increased lipolysis, and insulin resistance . The pheInsulin resistance can also be mimicked in 3D adipose tissue models. ASCs were co-cultivated with HUVECs after adipocyte differentiation in a 3D silk scaffold. Only co-culture ASCs differentiated into adipocytes showed a decrease in triglycerides content which may suggest an increase in lipolysis . This stBesides the relevance of endothelial cells, macrophage has a central role in the chronic inflammatory scenario established in adipose tissue in obesity ,14. A reIn the chronic inflammation scenario established in an obese adipose tissue, the long-term presence of proinflammatory macrophages leads to an unsolved fibrosis microenvironment . RajangaOrganoids have transformed the field of personalized medicine mainly due to their potential for predicting cellular and molecular events involved in the development of diseases ,112.The SVF fraction comprises a heterogeneous cell population, showing the main cellular elements of SAT such as stem cells, endothelial and immune cells. In adipose tissue engineering, the organoid concept emerged with the use of SVF to form 3D constructs to recapitulate adipose tissue morphology and physiology, with a particular attention to vascular structures and adipocyte-like cells showing unilocular lipid vacuoles ,113 FigA. In thiIn both studies, an external support (magnetic beads and Matrigel) was needed to maintain a 3D structure derived from SVF, distinct from ASC spheroids that secrete extracellular matrix components capable of sustaining their spheroidal morphology. Furthermore, these studies only showed functional improvements related to adipogenesis and angiogenesis ,113. DaqInterestingly, rat adipocyte progenitors were capable of migrating out of superficial fascia and generating adipose tissue organoids embedded in 3D hydrogel. These adipose tissue organoids derived from superficial fascia showed the majority of unilocular cells together with adipocyte functions involved with triglycerides metabolism and adipokine secretion . One impOther studies using human ASC or SVF cells combined with human-derived hydrogels showed potential as a 3D in vitro model of adipose tissue ,118. DecTo conclude, human ASC organoids compared with SVF organoids show some advantages including, but not restricted to: (1) ease of manipulation mainly due to the absence of hydrogel; (2) possibility of cell expansion and creating biobanks as a previous step to the organoids. However, one important disadvantage relies on the absence of endothelial cells. In this sense, alternative induction protocols must be tested. More importantly, mainly due to representing epigenetic alterations accumulated at stem and progenitor cells during obesity development, it would be interesting to obtain ASC organoids derived from obese donors. Obese organoids hold the potential of recapitulating the main cellular and molecular alterations which occurred during obesity development.Adipose tissue obese organoids will represent a powerful tool to modeling obesity besides testing potential drugs. However, besides their potential to revolutionize anti-obese drug discovery, organoids face challenges in their limited scale of production, absence of automation, costs, and reproducibility  . In addiThe diversity of WAT is reflected in its population of ASCs according to the adipose tissue physiology of each depot. Distinct layers of SAT reveal distinct ASC subpopulations and a possible integration with different subpopulations of fibroblasts located at the dermis, contributing together to skin and adipose tissue physiology. Recent advances in adipose tissue organoids opened an avenue of possibilities to recreate the main cellular and molecular events of obesity leading to a deep understanding of this inflammatory disease besides contributing to drug discovery. Obese organoids models will be more reliable when ASCs or SVF derived from obese donors are used, adding a personalized medicine context. Furthermore, 3D organ-on-a-chip will add reproducibility to these adipose tissue models contributing to their translation to the pharmaceutical industry.


Thus, here we provide a brief overview of the characteristics and physiological functions of the various types of NEVs, focusing on their production, isolation, functional characterization, and engineering strategies for their promising application as a cell-free modality for tumor immunotherapy.Extracellular vesicles (EVs) are increasingly recognized as important intermediaries of intercellular communication. They have significant roles in many physiological and pathological processes and show great promise as novel biomarkers of disease, therapeutic agents, and drug delivery tools. Existing studies have shown that natural killer cell-derived EVs (NEVs) can directly kill tumor cells and participate in the crosstalk of immune cells in the tumor microenvironment. NEVs own identical cytotoxic proteins, cytotoxic receptors, and cytokines as NK cells, which is the biological basis for their application in antitumor therapy. The nanoscale size and natural targeting property of NEVs enable precisely killing tumor cells. Moreover, endowing NEVs with a variety of fascinating capabilities  As the first line of defense against tumors and viral infections, NK cells can induce antigen-independent immune responses against malignant cells. A growing number of scientific reports and clinical studies have demonstrated that NK cell-based immunotherapy has promising antitumor effects . Furtherd tumors . Moreoved tumors . These f. Due to the above advantages, engineered NEVs have received a lot of attention to enhance their tumor-killing capabilities. Current research focuses on the use of engineered modifications to enhance the functionality of NEVs (Extracellular vesicles (EVs) can be divided into three subgroups based on their biological origin, including exosomes (30–150 nm in diameter), microvesicles , and apoptotic vesicles . The role of EVs secreted by immune cells in antitumor therapy has received more attention in recent years, with many studies confirming their great potential . Among t of NEVs . Neverth of NEVs .in vitro is not cytokine-dependent). It is inexpensive to administer, and there is substantial evidence that it is relatively safe  and cell lines are the two main sources of NK cells used in therapeutic studies. PBMC can be obtained from patient blood or blood collected from healthy adult volunteers. Among them, PBMC-induced NK cells from cancer patients had fewer clinical adverse effects and a higher safety profile . Howeverely safe . A studyely safe .Furthermore, umbilical cord blood (UCB), hematopoietic stem and progenitor cells (HSPCs), induced pluripotent stem cells (iPSCs), and CAR NK cells are valuable sources of NK cells . CAR-NK The most common method for isolating EVs is cell culture supernatant collection. Since there is no “gold standard protocol” for the preparation of pure EVs, the properties and functions of EVs may vary depending on the culture and isolation methods. Differences in cell culture oxygen content and inoculation surface have been shown to affect EVs production . DiffereObtaining EVs through natural secretion is hampered by the low yield. The advent of exosome-mimetic vesicles (EMs) with higher yield is expected to resolve this issue . Large-sEither artificially generated EMs or naturally secreted EVs typically contain multiple types of biological impurities. Therefore, it is essential to ensure that the purified products are inherently EVs without other contaminants before performing any functional analysis of the EVs. The current EVs isolation and purification methods and their advantages and disadvantages are summarized in As a promising cell-free therapy, achieving long-term and stable storage of NEVs plays a key role in their clinical application. Therefore, it is necessary to explore preservation techniques to protect the biological activity of NEVs for transport and clinical applications. Common conservation techniques include freezing, lyophilization, and spray drying . Any froIn recent years, as research on NEVs have continued, knowledge about the mechanisms underlying their function has been gained. Several studies have confirmed the ability of NEVs to target and kill tumor cells, which have been summarized in NEVs contain many substances acting as tumor killers, such as membrane proteins, toxic proteins, and miRNAs. In this section, we will elaborate on each section individually  are all found in NEVs . The perRegulatory miRNAs found in NEVs demonstrate tumor-killing and immunomodulation ability. Among these, miR-3607-3p encapsulated in NEVs inhibits cancer cell migration and invasion; miR-3607-3p-enriched NEVs may inhibit the malignant transformation of pancreatic cancer by directly targeting IL-26, and decreased miR-3607-3p levels were associated with poor prognosis and tumor metastasis . Anothervia the change of TAM -secreted cytokines and membrane surface proteins, and induces a direct antitumor effect of M1 macrophage  and enter the cancer reservoir to kill or deliver the drugs to central nervous system (CNS) tumors .Multiple studies have confirmed the ability of NEVs to target tumors . This chNEVs enter target cells through micropinocytosis, and the number of internalized NEVs correlates with tumor cell cytotoxicity . It was in vitro experiments, safety issues will become an issue that has to be addressed for the future development of NEVs.All current NEVs research has addressed the safety of NEVs at the cellular or animal level. Some studies point out that NEVs derived from PBMC have cytolytic activity against cancer cells but not against normal resting PBMC cells or normal cells . FurtherEVs-secreting behavior of NK cells is independent of cell activation status . Howevervia common engineering strategies has become a crucial direction for much applied research. Common EVs engineering techniques can be classified as endogenous and exogenous modifications. The primary objective is to increase the targeting ability or transform them into drug carriers (Endowing EVs with a variety of fascinating capabilities carriers . Existinvia electroporation (in vitro and in vivo (NEVs are suitable for drug delivery systems (DDS) due to their strong penetration, antitumor activity, and natural targeting. The current strategy is transporting exogenous drugs into NEVs, with engineering methods mainly including electroporation, ultrasonication, extrusion, freeze-thaw cycles, and saponin treatment . NEVs weporation  , which is enriched in NEVs and successfully enhanced tumor-killing ability by inhibiting overexpression of BCL-2 in breast cancer , 4D Kab. A studyThis review highlights the challenges and potential of NEVs in cancer therapy, which has demonstrated tremendous advantages in recent years as an emerging cell-free therapy in cancer immunotherapy, including smaller size, greater tissue penetration, lower acquisition costs, and independence from inhibitory TAM compared to conventional NK cell therapies. The NEVs inherit the tumor-killing and natural targeting abilities of their parent cells. It is associated with relatively few immune side effects due to the absence of cellular involvement. Therefore, in addition to cell therapy, NEVs have the potential to play a crucial role in future tumor immunotherapy.Pseudomonas aeruginosa lung infection by promoting M1 macrophage polarization. This suggests that NEVs may play a protective role in inflammation, especially in diseases with an imbalanced M1/M2 macrophage ratio (69). As the in-depth functions of NEVs have not yet be investigated, following research may need to focus on the functional contained biomolecules and the critical roles in the immune regulation process.The role of NEVs is still poorly understood, and researchers continue to investigate it. One study showed that NK-92MI cells-derived EVs could inhibit TGF-β1-induced HSC proliferation and activation, preventing liver fibrosis by carrying miR-223 . AnotherDespite the large number of studies demonstrating the efficacy of NEVs in cancer therapy, the development of NEVs still faces significant challenges. Not only do NEVs face these challenges, but all therapeutic EVs developments must also overcome them. The first is the heterogeneity of EVs, which complicates quality control and hinders a comprehensive understanding of their function. The main reason comes from the cell source and isolation methods for the production of EVs. The optimal cell source and isolation method for EVs is still under investigation. Ultracentrifugation is the most widely-used EVs isolation method, which needs to be integrated with another isolation method to improve the separation purity. The second challenge is selecting designs that improve the cycling stability and the cytotoxicity of NEVs. As mentioned above, although various endogenous and exogenous modification methods are used, there is no effective method to improve the loading efficiency of bioactive molecules without compromising the integrity of EVs, and most modification methods may cause clustering. Furthermore, it is necessary to evaluate the need for these modifications and their improvement in therapeutic efficacy. A reliable method to determine whether the loaded EVs contain active molecules is still urgently needed . Finally


What characteristics of the parent-adolescent relationship are associated with young adult health outcomes?In this cohort study of more than 15 000 adolescents, higher levels of adolescent-reported parental warmth, parent-adolescent communication, time together, academic expectations, relationship and communication satisfaction, and maternal inductive discipline were associated with favorable outcomes in young adulthood.This study suggests that investments in improving the parent-adolescent relationship may be warranted, given robust links with long-term health outcomes. Studies linking the quality of parent-adolescent relationships with young adult health outcomes could inform investments to support these complex relationships.To evaluate whether consistently measured, modifiable characteristics of parent-adolescent relationships are associated with young adult health across multiple domains.This cohort study used data from waves I  and IV  of the US National Longitudinal Study of Adolescent to Adult Health. Of 20 745 adolescents enrolled in wave I, 15 701 of 19 560 who were eligible completed wave IV . Data analyses were conducted from February 2019 to November 2020.Parental warmth, parent-adolescent communication, time together, relationship and communication satisfaction, academic expectations, and maternal inductive discipline as reported at wave I by adolescent participants.Wave IV participant-reported self-rated health, depression, stress, optimism, nicotine dependence, substance abuse symptoms , unintended pregnancy, romantic relationship quality, physical violence, and alcohol-related injury. Separate regression models were run for mother-adolescent and father-adolescent relationships while controlling for age, biological sex, race and ethnicity, parental educational level, family structure, and child maltreatment experiences.A total of 10 744 participants  and 8214 participants  had valid sampling weights and complete data for mother-adolescent and father-adolescent relationship characteristics, respectively. Adolescents who reported higher levels of mother-adolescent warmth , communication , time together , academic expectations , relationship or communication satisfaction , and inductive discipline  reported significantly higher levels of self-rated general health in young adulthood. Adolescents who reported higher levels of father-adolescent warmth , communication , time together , academic expectations , and relationship satisfaction  also reported significantly higher levels of self-rated general health in young adulthood. Adolescents reporting higher levels of all exposures also reported significantly higher levels of optimism and romantic relationship quality in young adulthood  and lower levels of stress and depressive symptoms . Higher levels of parental warmth, time together, and relationship or communication satisfaction were significantly associated with lower levels of nicotine dependence  and substance abuse symptoms , as well as lower odds of unintended pregnancy . Patterns were less consistent for physical violence and alcohol-related injury. Characteristics of mother-adolescent and father-adolescent relationships were similarly associated with young adult outcomes.The findings of this cohort study suggest that adolescents’ positive perceptions of their relationships with their mothers and fathers are associated with a wide range of favorable outcomes in young adulthood. Investments in improving parent-adolescent relationships may have substantial benefits for young adult population health. This cohort study uses data from the US National Longitudinal Study of Adolescent to Adult Health to evaluate whether consistently measured, modifiable characteristics of parent-adolescent relationships are associated with young adult health across multiple domains. First, many studies are short-term and do not use diverse samples, which limits generalizability.14 Second, strategies for measuring dimensions of parent-adolescent relationships vary widely, which makes it challenging to identify specific characteristics to target for intervention development.16 Third, most studies focus on health outcomes within selected domains, which precludes a more holistic understanding of the importance of parent-adolescent relationships on adult health across domains.13 Fourth, few studies include a separate focus on mother-adolescent and father-adolescent relationships, which may have important differences.18The importance of parent-adolescent relationships on the health of adult populations may influence decisions about investments to expand support for these complex relationships. Research shows that characteristics of family relationships during adolescence are associated with later health outcomes including mental health, sexual health, substance use, cardiovascular risk, and overall general health.19 Our primary goals were to create a clearer understanding of associations between parent-adolescent relationship characteristics and the health of adult populations and to inform future research. Previous research using an attachment theory framework has shown that the quality of parent-child relationships is associated with the health of children and adolescents.20 Within the context of this framework, we used Add Health to examine associations of mother-adolescent and father-adolescent relationship characteristics with young adult health in the US across the domains of general health, mental health, sexual health, substance use, and injury.The National Longitudinal Study of Adolescent to Adult Health  provides an opportunity to help address these limitations by evaluating whether consistently measured, modifiable, specific characteristics of parent-adolescent relationships are associated with young adult health across multiple domains.19 This cohort has been followed up over time. Our study focused on young adult health outcomes in the third decade of life, using data from wave IV, which was completed in 2008-2009 .19 We conducted a secondary data analysis to quantify associations between wave I adolescent-reported characteristics of mother-adolescent and father-adolescent relationships and wave IV health behaviors or outcomes. This study followed the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) reporting guideline for cohort studies.21 The University of North Carolina institutional review board approved all original Add Health study procedures, and participants provided written informed consent. Parental consent was required for students to participate in the study. Unless otherwise directed by the school, passive consent forms were used, such that it was assumed that a parent granted permission unless they returned a signed form indicating otherwise. However, some schools did require active consent forms in which a parent returned a signed form granting permission. The Committees for the Protection of Human Subjects at Children’s Hospital of Philadelphia approved an exemption from review for this study because data were deidentified.Add Health enrolled a large, nationally representative sample of more than 20 745 in-school adolescents who were in the 7th to 12th grades during the 1994-1995 school year (wave I).The Box. Most of the time, your mother [father] is warm and loving toward you.How close do you feel to your mother [father]?How much do you think your mother [father] cares about you?Which of the things listed have you done with your mother [father] in the past 4 weeks?Talked about someone you’re dating, or a party you went toTalked about a personal problem you were havingTalked about your school work or gradesTalked about other things you’re doing in schoolWhich of the things listed have you done with your mother or father in the past 4 weeks?Gone shoppingPlayed a sportGone to a religious service or church-related eventGone to a movie, play, museum, concert, or sports eventWorked on a project for schoolHow disappointed would your mother [father] be if you did not graduate from college?How disappointed would your mother [father] be if you did not graduate from high school?Overall, you are satisfied with your relationship with your mother [father].You are satisfied with the way your mother [father] and you communicate with each other.When you do something wrong that is important, your mother talks about it with you and helps you understand why it is wrong.Self-rated healthDepressive symptomsStressOptimismRomantic relationship qualityUnintended pregnancySmoking cigarettesAlcoholMarijuanaOther drugsPhysical violenceAlcohol-related injuryaSee eAppendix 2 in All characteristics of parent-adolescent relationships were measured by adolescent report at wave I and are reported on a scale of 1 to 5 . Adolescents were asked about various aspects of their relationships with their residential parent figures . Parental warmth was measured using the mean score of 3 items about love, closeness, and caring. Extent of communication was measured using the sum of 4 items on talking about social activities, personal problems, and school, then transforming the value to a scale of 1 to 5  for ease of comparison with the other exposure measures. Time together was measured using the sum of 5 items about shared activities , which were transformed to a scale of 1 to 5 . Academic expectations (graduation from high school and college) were measured using the mean score of 2 items. Adolescent relationship and communication satisfaction with parents was measured using the mean of 2 items. Each of these measures was asked separately for mother-figure and father-figure relationships. The single-scale item for inductive discipline (explaining why the adolescent has done something wrong) was asked only about mothers.22 Perceived stress was measured using 4 items from the Cohen Perceived Stress instrument and summing responses .23 Optimism was measured using 4 items on a 1 to 5 scale and summing responses . Romantic relationship quality was calculated as the mean score of 6 items on a scale of 1 to 5 (where 1 indicates strongly disagree and 5 indicates strongly agree) from the Supporting Healthy Marriage instrument.24 History of unintended pregnancy was measured with a single yes or no item. Nicotine dependence was measured using 4 items from the Heavy Smoking Index and coded as a dichotomous outcome (yes or no).25 Symptoms of alcohol, cannabis, and other drug abuse were based on 5-item scales from the Diagnostic and Statistical Manual of Mental Disorders (Fourth Edition) and summed to create count variables of the number of presenting symptoms .26 History of physical violence and risk of alcohol-related injury were each measured with single items on scales of 0 to 3 and 0 to 2, respectively, and collapsed to create dichotomous variables (yes or no).All health behaviors and outcomes were measured by participant report at wave IV. Self-reported general health was measured with a single item on a scale of 1 to 5 (where 1 indicates poor and 5 indicates excellent). Depressive symptoms were measured using 5 items from the Center for Epidemiologic Studies Depression instrument and summing responses .29Covariates included biological sex, race and ethnicity , wave I parental educational level (as a proxy for socioeconomic status), wave IV age, wave I family structure, and child maltreatment experiences reported at wave III and wave IV. These covariates were selected because they have been shown to be associated with both parent-child relationships (exposure) and health (outcome).30 We used listwise deletion to address missing data. Although data were not missing completely at random , the literature suggests that listwise deletion works well with large samples that have a small amount of missing data, such as Add Health, and that listwise deletion often accommodates data that are not missing at random in regression contexts, better than multiple imputation or full information maximum likelihood.34Statistical analyses were conducted from February 2019 to November 2020. All statistical tests were 2-sided. Analyses were restricted to respondents who had valid sampling weights and complete data on all variables of interest. Separate models were run for mother-adolescent and father-adolescent relationships. All analyses used sampling weights and adjusted variance estimates for the Add Health complex survey design using the subpop command in Stata, version 15.9 (StataCorp LCC), to allow reporting of nationally representative estimates. The first step in the analysis was to create and measure the internal consistency of our parent-adolescent relationship domain variables. Next, regression models tested for associations between each of the parent-adolescent relationship domains and each of the outcomes while adjusting for all covariates. The Holm-Bonferroni method was used to report only statistically significant differences at the .05 level after correction for multiple tests.The type of regression used depended on the strategy used for outcome measurement. Logistic regressions were used for nicotine dependence diagnosis, unintended pregnancy, physical violence, and alcohol-related injury, for which results are presented as odds ratios (ORs). Negative binomial regressions were used for alcohol abuse symptoms, cannabis abuse symptoms, and other drug abuse symptoms, for which results are presented as incidence rate ratios (IRRs). Ordinary least-squares regressions were used for depression score, stress score, optimism score, romantic relationship quality, and self-rated health, for which the β coefficients represent the estimated mean change in the outcome for every 1-unit increase in the variable. See eAppendix 1 in Adolescents who reported higher levels of mother-adolescent warmth , communication , time together , academic expectations , relationship or communication satisfaction , and inductive discipline  reported significantly higher levels of self-rated general health 14 years later . AdolescAdolescents who reported higher levels of mother-adolescent warmth , time together , academic expectations , relationship or communication satisfaction , and inductive discipline  had significantly lower levels of depressive symptoms 14 years later . The extAdolescents who reported higher levels of mother-adolescent warmth , communication , time together , academic expectations , relationship or communication satisfaction , and inductive discipline  reported significantly higher romantic relationship quality 14 years later . AdolescAdolescents who reported higher levels of mother-adolescent warmth , time together , and relationship or communication satisfaction  were significantly less likely to report having had an unintended pregnancy by young adulthood . Extent Adolescents who reported higher levels of mother-adolescent warmth  and relationship or communication satisfaction  had significantly lower odds of nicotine dependence. Similarly, maternal warmth and relationship or communication satisfaction were associated with significantly lower incidence of alcohol abuse symptoms , cannabis abuse symptoms ; satisfaction: IRR, 0.77 ), and other drug use symptoms  14 years later . AdolescResults from adolescent report of father-adolescent relationship characteristics and substance use in young adulthood were similar . Father-No characteristics of mother-adolescent relationships were significantly associated with odds of experiencing physical violence 14 years later . AdolescA visual summary of these primary analyses displaying the associations of mother-adolescent and father-adolescent relationship characteristics with young adult health behaviors and outcomes, while controlling for all covariates, is provided in the eFigure in A summary of the sample of participants with responses to both the mother and father relationship items (n = 7783) can be found in eTable 1 in 20 we add to the literature suggesting links to health through the third decade of life. In a nationally representative sample, with a set of consistently defined measures of parent-adolescent relationship characteristics, we found adolescents’ perceptions of their relationships with parents were associated with general health, mental health, sexual health, and substance use in young adulthood. Adolescent perceptions of their relationships with their mothers and their fathers were similarly associated with young adult health outcomes, above and beyond associations with adolescent biological sex, race and ethnicity, socioeconomic status, family structure, and history of child maltreatment. Our results can help to inform specific strategies for improving complex parent-adolescent relationships that could be associated with young adult population health.Our findings extend existing science to show that specific modifiable characteristics of parent-adolescent relationships are linked to favorable health behaviors and outcomes across multiple health domains 14 years later. Aligned with developmental science supporting the importance of quality of parenting relationships in child and adolescent health,8 also used Add Health data to examine the association between family “connectedness” (using an averaged response provided for mother and father items) and young adult health. Their analyses found associations between adolescent perceptions of family connectedness and wave IV measures of sexual health and substance use, but not mental health. Chen and Harris,1 in Add Health analyses extended to wave V, found that higher family “cohesion” and lower levels of parent-child “conflict” were associated with lower risk of depressive symptoms throughout young adulthood into midlife. Variability in findings may be partially associated with differences in measurement strategies.9 Nonetheless, taken together, adolescents’ perception of the quality of their relationship with parents appears to have a long-lasting association with young adult health behaviors and outcomes.Patterns were consistent. Adolescents who reported warm, loving, close, caring relationships with their parents; spending more time with their parents; and high levels of satisfaction with their relationship and communication with parents reported having higher levels of general health, feelings of optimism, and quality of romantic relationships in young adulthood; they also reported lower levels of depressive symptoms, stress, nicotine dependence, and symptoms of substance abuse. We also found consistent favorable patterns associated with adolescents’ report of maternal inductive discipline. These findings are generally aligned with existing literature using less-specific measures for parent-adolescent relationship quality and narrower ranges of young adult outcomes. Steiner et al38There was some variability in patterns between parent-adolescent relationships and young adult outcomes. Adolescent reports of parental academic expectations were less consistently associated with young adult outcomes, highlighting that the associations with specific characteristics of parenting relationships may vary by health behavior or outcome. Even though adolescent-reported satisfaction with parent relationships and communication was consistently associated with young adult outcomes, reported extent of communication with parents was not, highlighting the complexity of behaviors such as communication, and their measurement, within parent-adolescent relationships. Patterns were more consistent for some health-related outcomes and behaviors than others. For example, the links between quality of parent-adolescent relationships and emotional health in young adulthood seemed most consistent, while links with injury and physical violence outcomes were inconsistent or weaker, highlighting that health behaviors and outcomes are likely associated with many other factors.40Taken together, our results suggest that efforts to improve parent-adolescent relationship quality and adolescent perceptions of parent-adolescent relationships could have benefits for many  important young adult health behaviors and outcomes. Return on investment would go beyond benefits for adolescent health, reaching far into young adult populations and across a wide range of health domains. Building on existing and new research, it should be possible to identify and implement a portfolio of effective strategies to meet the needs of diverse parents and their adolescent children. Our results suggest that interventions focused specifically on influencing adolescents’ perceptions of relationship warmth, satisfaction with relationships and communication, time spent together, and inductive discipline should be prioritized. Although outreach is often to mothers, our findings suggest that interventions should include both mothers and fathers when feasible. Emerging research can inform such efforts, and more research is needed to explore innovative strategies, such as building peer support or mentoring father networks and mobile messaging interventions targeting fathers, to better engage fathers in adolescents’ health.43 Reviews of parenting interventions outside of clinic settings have shown favorable associations with the quality of parent-adolescent relationships, as well as health-related outcomes.47 Emerging literature shows that clinic-based interventions can effectively improve parenting communication and relationships as well as parent-adolescent communication about specific adolescent health-related outcomes in domains of sexual health, alcohol use, and teen driving.49 Our study supports the groundwork for a renewed interest in interventions and strategies designed to nurture and support the development of high-quality parent-adolescent relationships, above and beyond efforts targeting early childhood.50 Early-life factors associated with favorable quality of parent-adolescent relationships should be identified and supported, as well as strategies to “reset” low-quality relationships in late childhood and early adolescence when needed. All efforts to support parent-adolescent relationships must take into account that primary adult caregivers can look different across families.51 Diversity in personal values, cultural norms and expectations, family structure, and relationship dynamics need to be acknowledged and respected within the context of family relationships. There is a special need for more research on relationships between parents and sexual and gender minority youths.52Existing literature demonstrates effective strategies to influence parental expectancies and behaviors and favorably influence adolescents’ perceptions of parental trustworthiness, communication, and relationship satisfaction.This study has some strengths, including its large diverse sample, use of consistent specific measures for parent-adolescent relationships, and ability to test associations with a broad range of young adult health behaviors and outcomes. Analyses included adolescent relationships with both mothers and fathers.This study also has some limitations. It precluded detailed analyses related to specific characteristics of caregivers, relationships with nonresidential parents, and more specific outcomes. In the future, more granular analyses that consider nuanced complexities of adolescent gender identity, relationships with specific types of parental figures, family structure, and specific outcomes are warranted. Measures were limited to those included on Add Health surveys. This sample is representative of in-school adolescents in the 1994-1995 school year, and findings may not be generalizable to out-of-school youth, later cohorts, or individuals in other countries. We used adolescent-reported measures in this study, which likely differ from parental reports or perceptions. Multiple factors not measured in the Add Health study or included in our analyses contribute to the complex health behaviors and health outcomes used in our study, and results should be placed within this context.This cohort study produces a clearer understanding of the associations between specific characteristics of parent-adolescent relationships and the health of young adult populations that can inform future research. Adolescents’ perceptions of their relationships with parents are associated with a wide range of favorable outcomes in young adulthood. Investments in improving adolescents’ relationships with their mothers and fathers may have substantial benefits for young adult population health.


Editorial on the Research TopicCardiovascular sequelae of chemotherapy and radiotherapy in cancer survivors: current evidence and perspectivesCardiovascular diseases and cancer are the two leading causes of morbidity and premature death worldwide , 2. In tMoon-Sing Lee et al. in their population-based study, identified 158,798 breast cancer patients and using a propensity score match of 1:1, included 21,123 patients in each left and right breast radiation cohort. Previous heart disease and anticancer agents, including epirubicin, doxorubicin and trastuzumab, were taken into account for the analysis. Patients who received left radiation demonstrated an increased risk of ischemic heart disease compared with patients who received right breast radiation. Furthermore, in those undergoing left breast radiation dose >6,040 Gray, subsequent epirubicin might show a tendency to increase the risk of heart failure, whereas doxorubicin and trastuzumab did not. Further studies appear to be needed to confirm these hypothesis-generating results.Therefore, we should take into account the potential synergic effect of anticancer agents and radiotherapy –6. Moon-Torrente et al. conducted a single-institution retrospective study in order to assess the incidence and identify risk factors for cardiac events in patients treated with Immune checkpoint inhibitors (ICIs) for hematological and solid tumors. These agents produce a wide spectrum of inflammatory and immune-related adverse events  cells infusion for patients affected by advanced and refractory onco-hematological malignancies. CAR-T cell-related cardiovascular toxicities are now emerging as significantly affecting patients' prognosis. Mechanisms involved are still under investigation, although the aberrant inflammatory activation observed in cytokine release syndrome (CRS) seems to play a pivotal role. The most frequently reported cardiac events, observed both in adults and in the pediatric population, are represented by hypotension, arrhythmias and left ventricular systolic dysfunction, sometimes associated with overt heart failure. Therefore, there is an increasing need to understand the pathophysiological basis of these events and risk factors related to their development, in order to identify most vulnerable patients requiring a close cardiological monitoring and long-term follow-up. A multidisciplinary approach focused on the management of this selected population is now needed to optimize outcomes Wei-Ting Chang et al. starting from a reported increased risk of incident coronary heart disease and myocardial infarction in association with Gonadotropin Releasing Hormone (GnRH) antagonists treatment, aimed at studying whether regular aspirin use was associated with a risk reduction of major adverse cardiovascular and cerebrovascular events (MACCEs) by using a nationwide database. In their study, they observed that, despite a higher risk of MACCEs among irregular aspirin users, after adjusting for age, cancer stage and comorbidities, there was a trend to MACCEs reduction among those who received aspirin regularly.The inflammatory hypothesis may be also applied to the therapeutic field. Aspirin, through its anti-inflammatory effect, can inhibit tumor cell growth and prevent the development of cardiovascular diseases . By suppin vivo and potentially provide additional insights beyond functional evaluation -sequencing data. Mice were injected intraperitoneally with doxorubicin [ or ] to simulate acute and chronic cardiotoxicity. Left ventricular myocardium samples were analyzed by RNA-sequencing to identify differentially-expressed genes. Alas1, Atp5g1, and Ptgds revealed to be ideal biomarkers in acute cardiotoxicity, while Hsph1 and Vegfa were identified as potential biomarkers of chronic toxicity. This report first provided bioinformatics and clinical evidence for differences in mechanisms of doxorubicin-induced acute and chronic cardiotoxicity.Rapid development of microarray and sequencing technologies has revolutionized the complexity of collecting and examining molecular data in current biomedical research. Bei Chen et al. in the first large population-based study on the risk of fatal cardiac events in sarcoma patients, suggested that the risk of death is higher than that observed in the general population, and progressively increased with longer follow-up times. However, in patients with bone and soft tissue sarcoma, the risk of cardiac death varied mainly in patients with different histological subtypes of sarcoma and disease stages. Subgroup analyses indicated that chemotherapy increased the risk of events in patients with localized osteosarcoma, but not in those with other histological sarcoma subtypes and clinical characteristics. Thus, to mitigate the risk of death in sarcoma patients, enhanced multidisciplinary cooperation is warranted.The increasing need to understand the pathophysiological basis of cardiotoxicity and its early detection serves to identify most vulnerable patients requiring a close cardiological monitoring and, above all, a long-term follow-up . With imXuezhen Wang et al. explored the impact of chemotherapy on the risk of cardiac-related death in astrocytoma patients, by retrospectively evaluating astrocytoma patients diagnosed between 1975 and 2016 in the Surveillance, Epidemiology, and End Results (SEER) database. They found that age at diagnosis plays a crucial role in determining the risk of subsequent cardiac death rather than chemotherapy, highlighting that cardio–oncologists must provide comprehensive care and long-term monitoring for cancer patients, especially those at an increased cardiovascular risk.Similarly, This special issue again sheds light on the emerging role of cardio-oncology teams, always more necessary to reduce the burden of cardiovascular diseases in cancer patients through surveillance strategies implementation and cardiotoxicity management protocols.


Hand gesture recognition is a vital means of communication to convey information between humans and machines. We propose a novel model for hand gesture recognition based on computer vision methods and compare results based on images with complex scenes. While extracting skin color information is an efficient method to determine hand regions, complicated image backgrounds adversely affect recognizing the exact area of the hand shape. Some valuable features like saliency maps, histogram of oriented gradients (HOG), Canny edge detection, and skin color help us maximize the accuracy of hand shape recognition. Considering these features, we proposed an efficient hand posture detection model that improves the test accuracy results to over 99% on the NUS Hand Posture Dataset II and more than 97% on the hand gesture dataset with different challenging backgrounds. In addition, we added noise to around 60% of our datasets. Replicating our experiment, we achieved more than 98% and nearly 97% accuracy on NUS and hand gesture datasets, respectively. Experiments illustrate that the saliency method with HOG has stable performance for a wide range of images with complex backgrounds having varied hand colors and sizes. Gestures are used for human interaction to express feelings, communicate non-verbal information, and increase the value of messages. A gesture can be an intuitive human–computer interface that helps machines understand body language for various purposes. Both online and offline applications, such as interacting with a computer, recognizing pedestrians and police hand signs in automated cars, gesture-based game control, and medical operations that use this technology are still in their infancy.Two main approaches to detecting hand gestures are glove-based analysis and vision-based analysis. Glove-based techniques take advantage of sensors attached directly to the glove and accurately analyze hand movements. Vision-based methods can help users feel more comfortable without annoying physical limitations. They utilize a camera(s) to capture human hand signs and provide a more natural posture. The most essential ability of vision-based techniques is filtering out irrelevant and complex information and considering the most useful information during detection.In this paper, we propose a general method of hand gesture recognition based on computer vision methods and compare the empirical results of input images with complex backgrounds. Recognizing different hand signs using an integrated structure based on saliency maps and histogram of oriented gradients (HOG) creates a filter for selecting target regions by ignoring irrelevant information. This leads to an increase in the performance of gesture recognition algorithms. These methods detect the exact regions of hand gestures and ignore complex backgrounds from input images. Lastly, we improve a convolutional neural network (CNN) with two blocks to identify hand postures for increasing accuracy and stability. We use the NUS Hand Posture Dataset II and the hand gesture dataset to demonstrate the performance of our model. In our experiments, we applied six diverse aggressive types of noise such as Gaussian, impulse, Laplacian, multiplicative-Gaussian, Poisson, and uniform to around 60% of our datasets to evaluate our model’s performance while encountering low-quality images.The remainder of this paper is organized as follows. Ajallooeian et al.  used a sZamani and Rashidy  after exnd Davis  developend Davis  trained Vishwakarma et al.  detectedYang et al.  proposedach area .A two-stage hand gesture recognition is proposed to support a patient assistant system. The first step utilizes a saliency map to simplify hand gesture detection, and the second step classifies the patient’s postures. A novel combined loss function and a kernel-based channel attention layer are used to optimize the saliency detection model and emphasize salient features, respectively . Guo et Ma et al.  designedWe introduce a method that eliminates the complexity of image backgrounds using features extracted from original images and binary operators. Detecting objects in complicated scenes is one of the challenging tasks in hand gesture recognition since it is difficult to recognize the intent object among many others. The proposed model provides an efficient system based on deep learning for recognizing the structure of hand postures in complex backgrounds by developing the architecture shown in In this architecture, the size of the input image is equal to In this section, experiments are designed to evaluate the performance of the saliency map incorporating the HOG features.As can be seen in This study uses Python 3.7.12 with CUDA version 11.8.89 for all our experiments. The experiments have been carried out using PyTorch, an open-source and optimized tensor library for deep learning . The modIn the experiments, four main features, namely saliency map, skin color, histogram of oriented gradients (HOG), and Canny edge detection, are extracted from the main input image. We apply some aggressive image noises randomly in 60% of our datasets to alleviate the problem of insufficient training and testing data. The six types of image noises applied to our datasets are namely Gaussian, impulse, Laplacian, multiplicative-Gaussian, Poisson, and uniform. The real rate of noise distribution in all functions is 0.9 except for impulse, which is equal to 0.1  Figure 7Some of the referenced-based image quality estimation metrics such as mean square error (MSE), global relative error (ERGAS), multi-scale structural similarity index (MSSSIM), peak signal-to-noise ratio (PSNR), root mean squared error (RMSE), spectral angle mapper (SAM), structural similarity index (SSIM), universal quality image index (UQI), and visual information fidelity (VIF) are estimated for six different image noises mentioned above  which are applied on both the NUS Hand Posture Dataset II  and the We compared the proposed method with three state-of-the-art pyramid pooling and saliency detection methods, which include CNN-SPP  and saliChuang  proposedA hand-gesture-controlled PAS proposed in  uses a tThe NUS hand posture images with varying backgrounds, hand sizes, and skin colors are very challenging hand postures to identify. As can be seen in We introduced a novel method integrating the histogram of oriented gradients (HOG), skin color, Canny edge detection, and saliency maps using bitwise operators to detect hand postures with complex scenes by an improved CNN model. Using integrated feature maps identified the exact regions of the gestures in each input image and increased the accuracy. Apart from this, the proposed method enabled distinguishing postures better given complex backgrounds. The NUS hand posture II and the hand gesture datasets were used in the experiment, and the results showed that the proposed method improved the performance of hand gesture recognition in these datasets with and without image noises.In our future work, we will address issues with the quality evaluation of image dehazing methods in vision-based hand gesture recognition systems. The image quality of experiments (QoE) is an essential aspect of various intelligent systems like those detecting hand postures since low-quality images or videos can have an adverse effect on identification performance. Evaluating gesture detection by incorporating audio-visual saliency will be considered in the next step of recognizing hand gestures.


Phase separation (PS) is an important mechanism underlying the formation of biomolecular condensates. Physiological condensates are associated with numerous biological processes, such as transcription, immunity, signaling, and synaptic transmission. Changes in particular amino acids or segments can disturb the protein’s phase behavior and interactions with other biomolecules in condensates. It is thus presumed that variations in the phase-separating-prone domains can significantly impact the properties and functions of condensates. The dysfunction of condensates contributes to a number of pathological processes. Pharmacological perturbation of these condensates is proposed as a promising way to restore physiological states. In this review, we characterize the variations observed in PS proteins that lead to aberrant biomolecular compartmentalization. We also showcase recent advancements in bioinformatics of membraneless organelles (MLOs), focusing on available databases useful for screening PS proteins and describing endogenous condensates, guiding researchers to seek the underlying pathogenic mechanisms of biomolecular condensates. Biomolecular condensates organize cellular components in a temporally and spatially appropriate manner. It has been shown that the formation of some MLOs is initiated by liquid-liquid phase separation (LLPS) of particular proteins –3 . TheDysfunctional condensation is implicated in a variety of pathological processes linked to liquid- or solid-like condensates. For example, mutations of methyl CpG binding protein 2 (MeCP2) can disrupt heterochromatin assembly, which forms via LLPS in native status and develops into transcriptional dysregulation in Rett syndromeIn this review, we first characterize four types of sequence variations associated with dysfunctional condensates. We then provide a collective assessment of current computational resources for describing proteins driving liquid-like PS or solid-like phase transition, as well as their relations with disease-associated condensates.Theories that try to explain PS proteins in a universal mechanism often emphasize the importance of multivalency ,21 . OnStickers can be distinguished by employing a series of computational ,26 , thsat) and thus expand the PS propensity, similar to the effects of modulating the aromatic residues in hnRNPA1-LCDThe driving force for PS is governed by three factors, including the effective valence of stickers, the interaction strengths between stickers, and the patterning of spacers in the protein sequence ,27 . Thet al.et al.It has been reported that amino acids with similar chemical properties can demonstrate various sticker strengths. Bremeres) of PS proteinses stabilizes the PS process of A1-LCD, a homolog of hnRNPA1, when the Ser content is increasedes by increasing the charged contentet al.Spacers affect the phase behaviors and material properties of condensates by influencing the effective solvation volume mutBTB/BACK. The mutant tends to diffuse in the nucleus and is unable to localize to SPOP-DAXX bodies in HeLa cellsMissense mutations are a critical type of variation observed in proteins that affect biomolecular assembly. To obtain the overall landscape of functional mutations of PS proteins, we referred to PhaSepDB, which provides a collection of manually curated PS-related proteinses, according to the stickers-spacers modelde novo design for manipulating PS.In addition to the disorder status and frequency of particular mutations, the physiochemical properties of drivers are relatively preserved in the regulation of PS. Previous study shows that mutating attractive groups into hydrophilic amino acids partially accounts for the loss of PS propensity. For example, the W131G mutation is expected to have an impact on SPOP phase behavior by increasing its vet al.et al.Expansion of sequence repeats is another key type of disease-associated variation that occurs in dozens of inherited human disorders ,102 . PFusion genes are a crucial category of cancer drivers that are identified in approximately 20% of cancer morbidityThe length and amino acid composition of attractive groups in PS proteins can also be modulated pretranslationally through AS, which produces distinct mature mRNAs from a single pre-mRNA by including or skipping different exon segments. Approximately 95% of multiexon genes are expected to undergo AS –124 , me.g., the splicing of FXR1 in the development of Xenopus. Smithet al.AS of Ser- and Arg-rich IDRs can achieve different morphologies of condensates,Previous studies discovered that mammalian exons with specific AS events are enriched in IDRs containing Gly- and Tyr-rich motifs, such as hnRNP families and other RBPs, which play diverse roles in RNA processinget al.It has been observed that AS regulates the assembly properties of RNA-processing proteins by manipulating the incorporation of IDRs in different isoformsFailure in the regulation of material states may lead to aberrant protein assemblies, which could trigger a range of pathological processes, with the sequence variations mentioned above as a possible prominent mechanism –147 . TSeveral LLPS-associated protein and RNA databases were released until 2023. Since the original publications and web interfaces of these resources have already provided detailed explanations on their usage, we aim to provide a guide for users on the strengths of these resources and explain particular usage where they could be beneficial. The main features of the databases discussed are summarized in Tablesi.e., scaffolds and regulators, which were assigned to 40 biomolecular condensates. This database also documents PS-associated proteins in 164 eukaryotes through computational prediction. LLPSDB is a web-accessible database providing curated LLPS proteins and correspondingin vitro experimental conditions from published literaturein vivo orin vitro. PhaSepDB is a suitable resource for those who want to gain an overview ofin vitro andin vivo experiments performed in proof of LLPS of a target protein.DrLLPS is an integrative database for the proteins that can undergo LLPSIncreasing evidence has demonstrated the essential roles of RNAs in the functions and maintenance of MLOs such as nucleoli, P-bodies, and stress granules. At present, two resources have addressed and collected RNAs related to PS and condensates . RNAPhaSA range of computational algorithms have been developed to predict PS proteins, which are to some extent applicable for proteins with higher contents of IDRs or LCDs. PScore predicts long-range π-π interactions based on statistically expected interactions between π-orbital-containing residuesMost current predictors only perform well in proteins that can self-assemble to undergo PS (SaPS). PhaSePred is a machine-learning predictor that can predict the potential of proteins to assemble through partners, considered partner-dependent PS (PdPS) proteinsThe discovery and prediction of protein constituents in biological condensates present enormous opportunities for uncovering underlying mechanisms. The DeepPhase method provides a global view of the human PS proteome, according to the morphology of their condensates, instead of using sequence-dependent traits such as other predicting toolse.g., partner-dependent PS proteins and regulators. Prediction efficiency depends on standard negative and positive phase separating protein datasets, which are thus far limited. Indeed, these predictors successfully predict numerous phase-separating proteins, confirming the importance of phase-separation-prone features and providing the foundation to establish more integrative and accurate predictors or tools for protein science in the future [Informative reviews have comprehensively summarized the route performance and primary principle, which suggests a limited overlap between existing predictors that may generate a significant rate of false negatives –164 . T future ,165 .It is clear that PS is presumed to play ubiquitous roles in fundamental biological processes. The question is thus raised as to whether PS is a requisite in a given function. A detailed experimental pipeline for LLPS-associated studies has been provided by Gao and coworkers, addressing how we can relate the experimental phenomena to biological functionsIncreasing numbers of useful bioinformatics methods have been developed to predict PS proteins. However, most of these methods are designed for screening such proteins with higher IDR contentA range of diseases are associated with gene mutations, which can lead to the gain or loss of the ability to condense during the development of pathogenesis ,170 . F