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Background: The quality of early experiences with caregivers affects individual adjustment and can modulate adults' responses to salient social stimuli, like infant faces. However, in the framework of Interpersonal Acceptance-Rejection Theory (IPARTheory), no research to date has examined whether early experiences of acceptance or rejection from caregivers are associated with individual differences in the electrophysiological (EEG) responses to infant faces., Objective: This study examined the associations between the perceived quality of care during childhood and the behavioral and EEG responses to infant and adult faces in non-parent young adults., Methods: N = 60 non-parent young adults (30 males; 30 females) completed an Emotion Recognition task displaying emotional and unemotional infant and adult faces during an EEG recording. Memories of past care experiences with mothers and fathers were collected using the short form version of the Parental Acceptance-Rejection scale., Results: At the behavioral level, slower Reaction Times (RTs) in recognizing all faces were related to higher levels of perceived maternal rejection in young adults; in particular, males who reported higher levels of maternal rejection displayed longer RTs in recognizing faces compared to females. At the neurophysiological level, as the level of perceived paternal rejection increased, the N170 amplitude to infant faces increased. Females who reported higher levels of paternal rejection, compared to males, had a larger increase in the N170 amplitude and a larger decrease in the LPP amplitude in response to emotional faces., Conclusions: While a higher perception of maternal rejection hindered the behavioral responses of adults in recognizing faces, those who felt more rejected by their own father during childhood showed an enhanced N170 amplitude to infant faces. This might reflect a greater need for discrimination resources, at a very early stage of infant face processing, in those adults who perceived higher levels of paternal rejection. Adults' sex modulated the associations found at the behavioral and neurophysiological levels. Overall, our findings extended the IPARTheory postulates that being neglected during childhood might trigger perceptual changes in adults, hindering the elaboration of social cues like infant and adult faces at different levels., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

While research has shown that the attentional bias to infant faces predicts the quality of infant care, the antecedents of this cognitive process have been less established. In particular, it remains unknown whether the attentional bias to infant faces might be affected by the interplay between different factors, including memories of past experiences of care, adults' sex, and the experience of parenthood. To extend previous results, we examined the attentional bias to infant faces in a mixed sample of parents ( n = 99) and nonparents ( n = 102), and whether it varied in relation to parental status, sex, the quality of past experiences of care, and the interactions between these factors. A modified go/no-go task was used to compare the effect of adult and infant faces in retaining adults' attention. Memories of past experiences of paternal and maternal care were collected using the short form of the Italian version of the Parental Acceptance-Rejection scale. Results confirmed that infant faces induced greater attentional interference compared to adult faces. A heightened attention to all types of faces was found in parents versus nonparent. Women, as compared to men, were slower in the task performance, and allocated more attention to infant versus adult faces. Consistent with previous evidence, the attentional prioritization of infant faces varied in relation to previous experiences of care with one's own mother; individuals who remembered a more accepting maternal care allocated more attention to infant versus adult faces. Parental status did not modulate this effect, but sex of participants did. Grounded in the interpersonal acceptance-rejection theory (IPARTheory), this study provides new insights for discerning processes that might regulate global adult caregiving. (PsycInfo Database Record (c) 2024 APA, all rights reserved).

Purpose: Recruitment and activation of inflammatory cells, such as retinal microglia/macrophages, in the subretinal space contribute significantly to the pathogenesis of age-related macular degeneration (AMD). This study aims to explore the functional role of vascular endothelial growth factor (VEGF-A), placental growth factor (PlGF) and VEGF-A/PlGF heterodimer in immune homeostasis and activation during pathological laser-induced choroidal neovascularization (CNV)., Methods: To investigate these roles, we utilized the PlGF-DE knockin (KI) mouse model, which is the full functional knockout (KO) of PlGF. In this model, mice express a variant of PlGF, named PlGF-DE, that is unable to bind and activate VEGFR-1 but can still form heterodimer with VEGF-A., Results: Our findings demonstrate that, although there is no difference in healthy conditions, PlGF-DE-KI mice exhibit decreased microglia reactivity and reduced recruitment of both microglia and monocyte-macrophages, compared to wild-type mice during laser-induced CNV. This impairment is associated with a reduction in VEGF receptor 1 (VEGFR-1) phosphorylation in the retinae of PlGF-DE-KI mice compared to C57Bl6/J mice. Corroborating these data, intravitreal delivery of PlGF or VEGF-A/PlGF heterodimer in PlGF-DE-KI mice rescued the immune cell response at the early phase of CNV compared to VEGF-A delivery., Conclusions: In summary, our study suggests that targeting PlGF and the VEGF-A/PlGF heterodimer, thereby preventing VEGFR-1 activation, could represent a potential therapeutic approach for the management of inflammatory processes in diseases such as AMD.

An attentional bias toward infant versus adult faces has been detected in parents and positively associated with sensitive caregiving behaviors. In previous research, the attentional bias has been measured as the difference in attention, in terms of reaction times, captured by infant versus adult faces; the larger the difference, the greater the cognitive engagement that adults deployed to infant faces. However, research so far has been mostly confined to samples of mothers, who have been more represented than fathers. Moreover, new family forms, especially same-sex families of men, have been left out of research. To clarify potential sex differences and extend previous findings to diverse family forms, we implemented a modified Go/no-Go attentional task measuring attentional bias to infant faces in parents with children aged from 2 to 36 months. The sample (N = 86) was matched and included 22 fathers and 22 mothers from different-sex families and 20 fathers and 22 mothers from same-sex families. Overall, the results confirmed that infant faces induced a greater attentional bias compared to adult faces. Moreover, we found that neither the type of family nor parents' sex modulated the attentional bias toward infant faces. The findings are discussed in relation to the importance of understanding the correlates of parental response to infant cues going beyond a heteronormative perspective on parenting., (© 2024. The Author(s).)

Purpose: We aimed at assessing the correlation between TEG reaction time (TEG-R) in citrated and fresh blood samples with TEG5000 and TEG 6S during heparin administration in patients with and without ECMO support., Materials and Methods: Paired TEG5000 (fresh and citrated whole blood, kaolin and kaolin-heparinase) and TEG6S (citrated whole blood) samples were obtained, together with standard coagulation laboratory tests. Bland-Altman analysis and Lin's concordance correlation coefficient were used to assess agreement., Results: Thirteen consecutive ECMO patients and eight consecutive non-ECMO patients were enrolled and TEG was performed for a total of 84 paired samples. ECMO patients received 19.2 (12.6-25.8) U/kg/h of heparin. Five of the non-ECMO patients did not receive heparin, two of them received a very low prophylactic dose (1.6 and 2.9 IU/kg/h, respectively), and one of them 13.1 U/kg/h of heparin. Using TEG®5000, TEG-R was 21.0 (-23.4; 65.5) min longer on fresh compared to citrated blood in patients receiving heparin while only 1.58 (-5.5; 8.7) min longer in patients not-receiving heparin. These differences were reverted by heparinase., Conclusions: Using citrated-recalcified blood to perform TEG might lead to underestimation of the effect of heparin., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Introduction: Extracorporeal membrane oxygenation (ECMO) may act as a driver or propagator of systemic inflammation. In turn, cytokine release can modify thromboelastographic (TEG) tests which are commonly used for anticoagulation monitoring. In this context, antithrombin (AT) supplementation might further modify TEG., Methods: This is a pre-specified sub-study of the "Randomized Controlled Trial of Antithrombin Supplementation During Extracorporeal Membrane Oxygenation" study (investigator-initiated, randomized, single-blind, two-arm trial) conducted in two Italian ECMO referral ICUs. Adult patients requiring vv-ECMO for respiratory failure and undergoing unfractioned heparin (UFH) administration were enrolled and randomized whether to receive AT supplementation. Plasma samples for cytokine assay (IL-8, IL-10, IL-6, IL-1β, TNF-α and Pro-ADM) and heparinase TEG were collected from every patient before ECMO start, 24 h and 72 h after ECMO start, before ECMO removal, and 7 days after ECMO removal or upon ICU discharge whichever happened first. AT concentration, coagulation and clinical data were collected before ECMO start and at pre-fixed time points., Results: Thirty-nine patients were enrolled (21 treatments, 18 controls). TEG-R had a weak-to-moderate positive correlation with IL-8, IL-6, IL-10 and TNF-α and a moderate positive correlation with Pro-ADM. TEG-ANG showed a weak negative correlation with IL-8, IL-6 and TNF-α, while TEG-MA negatively correlated with IL-8, TNF-α and Pro-ADM. AT supplementation seemed to modify the association between TEG-MA and IL-8, IL-10 and Pro-ADM; conversely, AT did not affect the relationship among TEG-R or TEG-ANG and the studied cytokines., Conclusions: High concentrations of systemic cytokines correlated with longer reaction times and decreased angle and amplitude at TEG, suggesting that an increase in inflammation is related with hypocoagulability as revealed by thromboelastography., Competing Interests: Declaration of conflicting interestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Inflammation plays a crucial role in cancer progression, but the relevance of the inflammasome remains unclear. Alu RNA was the first endogenous nucleic acid shown to activate the NLRP3 (nucleotide-binding domain leucine-rich repeat containing 3) inflammasome. Here, we showed that Alu RNA can induce epithelial-to-mesenchymal transition (EMT) through NLRP3 inflammasome activation and IL-1β release in colorectal cancer (CRC) cells. Alu RNA is stored, transported and transferred to CRC cells by exosomes. Exosomal Alu RNA promotes tumorigenesis by inducing invasion, metastasis and EMT via NLRP3 inflammasome activation. Consistent with these data, we found that significantly increased Alu RNA expression correlates with the induction of NLRP3 priming in human CRC patients. Furthermore, the level of Alu RNA in circulating exosomes correlates with CRC progression in a preclinical model. These findings reveal the direct involvement of Alu RNA in cancer pathogenesis, and its presence in CRC cell-derived exosomes could be used as a noninvasive diagnostic biomarker., (© 2024. The Author(s).)

In the era of immunotherapy, the targeting of disease-specific biomarkers goes hand in hand with the development of highly selective antibody-based reagents having optimal pharmacological/toxicological profiles. One interesting and debated biomaker for several types of cancers is the onco-fetal protein Cripto-1 that is selectively expressed in many solid tumours and has been actively investigated as potential theranostic target. Starting from previously described anti-CFC/Cripto-1 murine monoclonal antibodies, we have moved forward to prepare the humanized recombinant Fabs which have been engineered so as to bear an MTGase site useful for a one-step site-specific labelling. The purified and bioconjugated molecules have been extensively characterized and tested on Cripto-1-positive cancer cells through in vitro binding assays. These recombinant Fab fragments recognize the target antigen in its native form on intact cells suggesting that they can be further developed as reagents for detecting Cripto-1 in theranostic settings., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: MENOTTI RUVO reports financial support was provided by Italy Ministry of University. MENOTTI RUVO reports financial support was provided by Campania Region. Maria Cantile, Riccardo Sanna reports financial support was provided by Italy Ministry of University. Maria Cantile and Riccardo Sanna are employees of BIOVIIIx srl., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Attentional bias toward infant faces is associated with parental sensitivity and supports the infant-caregiver attachment relationship, ultimately fostering child health outcomes. However, experience-related determinants of parents' attentional bias to infant faces have been poorly investigated. We examined attentional bias to infant versus adult faces in a sample of same-sex mothers ( N  = 76), and whether it varied depending on maternal involvement in childcare and the perceived quality of past experiences of care., Method: A Go/no-Go attentional task was used to compare the effects of infant and adult faces in retaining attention. Maternal involvement in childcare was measured using items addressing nurturing behaviors. Memories of past experiences of care were collected using the short-form version of the Parental Acceptance-Rejection scale., Results: Results confirmed that infant faces induced greater attentional bias compared to adult faces. More involved mothers were more biased, in terms of attention, to infant versus adult faces. Attentional bias to infant versus adult faces increased as mothers felt more rejected by their own fathers during childhood., Discussion: Our findings suggested that attentional bias to infant faces might be associated with past experiences of care and direct commitment in childcare in same-sex mothers. Robust and accurate empirical findings on same-sex parent families are essential to inform social policies supporting these families' well being., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 The Authors.)

Introduction: Verbal language is one of the most immediate and significant means parents have to express affect and information to their children. Parental speech directed to children has been thoroughly examined in typical development. However, the characteristics of parental speech directed to children with neurodevelopmental disorders are far less well documented, and no recent studies have been carried out that involve autistic 1 children and their fathers. Therefore, the present study aims to analyze and compare maternal and paternal speech directed to young autistic children, focusing on fathers' elements of speech in comparison with maternal language., Methods: N  = 88 dyads participated in this study. 44 autistic children (41 males and 3 females) (chronological age: M  = 40.01 months; SD = 11.96) in interaction with their fathers (paternal age M  = 41.84 years; SD = 7.02) and the same 44 children in interaction with their mothers (maternal age M  = 37.37 years; SD = 5.45). The language was verbatim transcribed using ELAN software (ELAN Version 6.4, 2022) and coded with an observational tool (Penman) for analyzing functions and referents of speech after reaching a satisfactory level of agreement between two independent transcribers., Results: No differences emerged considering the affective aspects of speech. However, mothers seem to direct more informative salient statements ( W  = 1,259; p  = 0.02) and call the child's attention more often than fathers ( W  = 1,253.5; p  = 0.02). Regarding referents of informative speech, fathers focused more on the child's internal states rather than mothers ( W  = 727; p  = 0.04)., Discussion: These results reveal that fathers seem to display a relationship-based approach focused on a non-intrusive style with few demands while talking with their children, providing a complementary role to mothers that allows complete and harmonious stimulation of all areas of child development., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Perzolli, Bentenuto, de Falco and Venuti.)

Mothers and fathers of autistic children (ASD) tend to report elevated levels of parenting stress. Thus, it is critically important to understand which factors contribute to an imbalance between the perceived demands of parenting and the available psychological resources. To date, little is known about the association between child attachment representations and parenting stress. In this study, we first examined group differences in parenting stress levels based on child diagnosis and parents' gender. Second, we explored the predictive role of child diagnosis, autism severity, and child attachment representations on parenting stress. The study involved 23 school-age children with ASD (IQ > 70), 27 without ASD (7-13 years), and their mothers ( n = 50) and fathers ( n = 50). Data were collected from 2017 to 2020. Parents completed the Social Responsiveness Scale 2 and the Parenting Stress Index-Short Form, while the children's attachment representations were assessed using the School-age Assessment of Attachment. Parents of children with ASD reported higher stress compared with controls. No differences were found between mothers and fathers. Implicit attachment representations have been found to be associated with parenting stress only in mothers, while the severity of social impairment showed a significant effect on parenting stress in both parents. These findings revealed the potential benefit of adaptive attachment representations not only for children themselves but also for mothers and the family system, suggesting the bidirectional nature of parent-child relationships in the context of ASD. The uniqueness of maternal and paternal parenting experiences should be considered when parenting stress is addressed.

Infant faces are prioritized by the attentional system in parents, resulting in a greater cognitive engagement in terms of response time. However, many biological, contextual and environmental factors relating to this cognitive mechanism have been left unexplored. To fill this gap, this study aims to (i) confirm that infant faces engage more attention compared to adult faces; (ii) investigate whether the attention to infant faces is affected early care experiences of parents; (iii) explore the effect of parents' sex by taking the amount of involvement with early childcare into consideration. 51 mothers and 46 fathers completed a modified Go/no-Go task, a brief sociodemographic questionnaire, the short version of the Adult Parental Acceptance-Rejection scale, and an ad-hoc question relating to the amount of parental involvement with early childcare. Parents' response times were slowed in the presence of infant versus adult faces. Parents whose mother was perceived as more sensitively accepting were more engaged by infant cues. By considering the amount of early parental involvement, the sex of parents did not significantly interact with the type of face. These findings provide new insights on the attention process in response to infant cues in parents and suggest that the investigation of experience-based factors may shed further light on this topic.

Research during the COVID-19 pandemic has shown a strong relationship between child symptoms, parental stress, and mental health challenges. The pandemic has changed family routines, worsening child symptomatology and parental burden. The aim of this study was to investigate how the magnitude of the perceived changes in child externalizing behavior, parental stress, and discontinuity of therapy-from before to during the COVID-19 pandemic-affected parental mental health during the pandemic. Moreover, we sought to compare these aspects cross-culturally between European countries and the USA. To these purposes, we asked Italian, Spanish, and U.S. parents of children with neurodevelopmental disabilities (NDD) to complete an online survey. Quantitative results showed that increased parental stress may have contributed to a worsening in parental psychological distress, regardless of culture. Moreover, they suggested an indirect effect of child externalizing behaviors on parents' psychological distress via parental stress. Qualitative analyses highlighted that the lack, or discontinuity, of therapeutic activities may have been one of the key contributors to parenting burden during the COVID-19 pandemic. Finally, qualitative results highlighted resilience factors that could have decreased the risk of psychological problems during the pandemic, such as a strong sense of parental efficacy and the ability to adapt to changing family dynamics.

In recent years, the advanced knowledge of clinical, biological and molecular features of prostate cancer have led to the introduction of new drugs and have allowed the relocation of old drugs in different settings. In this way, the new concepts of systemic disease arise: high risk or high volume vs. low risk and low volume disease castration sensitive prostate cancer (CSPC), diversifying the use of previously approved drugs (CRPC) and opening new scenarios for sequence therapy. The aim of this review is to integrate new developments into the medical management of systemic prostate cancer.

The few studies available on quality of attachment in school-age children with autism spectrum disorder (ASD) exclusively used questionnaires assessing explicit attachment representations. Thus, in the current study we assessed both explicit and implicit attachment representations in 23 children with ASD (without intellectual disability), 22 with learning disabilities and 27 with typical development aged from 7 to 13 years. A self-reported measure on the quality of attachment to parents and a semi-structured interview were administered to the children. In addition, a developmental assessment of the child including measures of intelligence and social-communication impairment was conducted. Despite the lack of group differences on explicit attachment representations, we found that children with ASD showed higher rates of at-risk self-protective strategies and psychological trauma compared to the TD group. Children with SLD also showed a high level of at-risk implicit attachment representations than TD, albeit to a lesser extent compared to children with ASD. These results may be related to several factors associated with ASD impairment and developmental pathways, such as the atypical learning process which occur at interpersonal level, the difficulties in social information processing and reflective functioning. Our findings suggested that children with ASD may experience difficulties in the construction of balanced implicit attachment representations. Thus, a more comprehensive assessment of attachment including both implicit and explicit representations is recommended.

The present study aims to investigate cross-cultural differences and similarities in maternal and children's adjustment to COVID-19 restrictions in Spain and Italy, with reference to determinants of maternal stress. A total of 950 mothers (45.4% from Spain, n = 432, M age = 39.6, years old, SD = 5.2 and 54.5% from Italy, n = 518, M age = 40.5, years, SD = 6.4) of children aged 3-17 years old completed questionnaires on sociodemographic and COVID-19 contextual factors, parenting-related variables, and children externalizing behaviors using an online survey. We examined determinants of parental stress in both countries. Mothers reported a significant increase in parental stress and child externalizing behaviors during COVID-19 home confinement, with more remarkable perceived changes in the Spanish group. Hierarchical linear regressions showed that child age and externalizing behaviors, maternal psychological distress, quality of coparenting and pre-COVID-19 levels of parental stress significantly predicted parental stress in both Italian and Spanish mothers during the pandemic. Results confirmed the negative psychological impact of COVID-19 home confinement on parents and children across countries and revealed common underlying factors responsible for parental stress in Italy and Spain. These findings highlighted the need for communitarian preventive programs to reduce mental health difficulties and parental stress in mothers and behavioral difficulties in children during pandemic restrictions. Particular attention should be paid to young mothers and children, and the key role of positive partner support and coparenting relationships should be considered. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Veno-venous Extracorporeal Membrane Oxygenation (ECMO) is used in the most severe cases of respiratory failure and further exacerbates the patients' inflammatory status. Antithrombin is supplemented during ECMO for its anticoagulant effects, but it also deploys anti-inflammatory properties. In this pre-specified ancillary study of the GATRA trial [NCT03208270] we aimed to evaluate the relationship between antithrombin and inflammation during ECMO. Forty-six patients were included in the study, 23 were randomized to receive antithrombin to maintain a level of 80-120% (study group) and 23 were randomized not to be supplemented (control group). Anticoagulation was provided in both groups with heparin infusion. Six cytokines were measured at 5 timepoints from prior to ECMO start to 7 days after ECMO removal. Cytokines decreased during the study but overall were not very different in the two groups. Testing the interaction between the study group and timepoints suggests that the administration of antithrombin led to a more rapid decrease over time of IL-6, IL-1β, TNF-⍺ and Pro-ADM. Plasma levels of antithrombin (either endogenous or exogenous) were negatively associated with all cytokines. Inflammation decreases during ECMO but a causal effect of antithrombin administration on the reduction of inflammation (and its clinical relevance) must be confirmed by appropriately powered studies., (© 2022. The Author(s).)

Evidence of psychological distress in families during COVID-19 outbreak are arising. However, the perceived changes in psychological adjustment during home confinement with respect to the period before the pandemic have not been addressed yet. Moreover, little is known about the role of coparenting and specific COVID-19 contextual variables on parental stress and children's behavioral difficulties in the Italian context. Using a cross-sectional survey, we collected data on 841 Italian parents of children aged 3-11 years with typical development during the home confinement (20th April-18th May). We analyzed levels of parental stress, coparenting, and child externalizing behaviors before and during the home confinement. Additionally, hierarchical regressions were performed to investigate predictors of parental stress and child externalizing behaviors during the lockdown. Results showed that parental stress (especially in mothers) and child externalizing behaviors increased during the lockdown period. Coparenting was a strong predictor of parental stress, together with being a mother, younger child age, less time dedicated to the child, and scarce feasibility of remote working. Besides, child externalizing behaviors were predicted by male gender, less parental time dedicated to the child, higher parental stress, and child distance learning workload. Our findings indicate a negative impact of COVID-19 lockdown in both parents and children, suggesting that positive coparenting and time dedicated to children may help to reduce the detrimental effect of pandemic restrictions on family adjustment., (© 2021 The Authors. Family Process published by Wiley Periodicals LLC on behalf of Family Process Institute.)

Objectives: Extracorporeal carbon dioxide removal is used to treat patients suffering from acute respiratory failure. However, the procedure is hampered by the high blood flow required to achieve a significant CO2 clearance. We aimed to develop an ultralow blood flow device to effectively remove CO2 combined with continuous renal replacement therapy (CRRT)., Design: Preclinical, proof-of-concept study., Setting: An extracorporeal circuit where 200 mL/min of blood flowed through a hemofilter connected to a closed-loop dialysate circuit. An ion-exchange resin acidified the dialysate upstream, a membrane lung to increase Pco2 and promote CO2 removal., Patients: Six, 38.7 ± 2.0-kg female pigs., Interventions: Different levels of acidification were tested (from 0 to 5 mEq/min). Two l/hr of postdilution CRRT were performed continuously. The respiratory rate was modified at each step to maintain arterial Pco2 at 50 mm Hg., Measurements and Main Results: Increasing acidification enhanced CO2 removal efficiency of the membrane lung from 30 ± 5 (0 mEq/min) up to 145 ± 8 mL/min (5 mEq/min), with a 483% increase, representing the 73% ± 7% of the total body CO2 production. Minute ventilation decreased accordingly from 6.5 ± 0.7 to 1.7 ± 0.5 L/min. No major side effects occurred, except for transient tachycardia episodes. As expected from the alveolar gas equation, the natural lung Pao2 dropped at increasing acidification steps, given the high dissociation between the oxygenation and CO2 removal capability of the device, thus Pao2 decreased., Conclusions: This new extracorporeal ion-exchange resin-based multiple-organ support device proved extremely high efficiency in CO2 removal and continuous renal support in a preclinical setting. Further studies are required before clinical implementation., Competing Interests: Drs. Zanella and Pesenti are inventors of a patent related to this study. Title: “Extracorporeal Circuit for Removal of CO2 from Blood”; application number: WO2019IB54288 20190523; applicants: Fondazione IRCCS Ca’ Granda – Ospedale Maggiore Policlinico, and Università degli Studi di Milano. Dr. De Falco received support for article research from the Italian Ministry of Health. Drs. De Falco, Gori, and Grasselli disclosed the off-label product use of the device for extracorporeal CO2 removal and renal replacement therapy. Dr. Di Girolamo disclosed the off-label product use of resins. Dr. Colombo disclosed government work. Drs. Gori’s and Grasselli’s institutions received funding from the Italian Ministry of Health. Dr. Pirrone received support for article research from the National Institutes of Health. Dr. Grasselli received funding from Fisher & Paykel, Dreager Medical, Biotest, Getinge, Thermofisher, and MSD. The remaining authors have disclosed that they do not have any potential conflicts of interest., (Copyright © 2022 by the Society of Critical Care Medicine and Wolters Kluwer Health, Inc. All Rights Reserved.)

The clinical outcome of the disease provoked by the SARS-CoV-2 infection, COVID-19, is largely due to the development of interstitial pneumonia accompanied by an Acute Respiratory Distress Syndrome (ARDS), often requiring ventilatory support therapy in Intensive Care Units (ICUs). Current epidemiologic evidence is demonstrating that the COVID-19 prognosis is significantly influenced by its acute complications. Among these, delirium figures as one of the most frequent and severe, especially in the emergency setting, where it shows a significantly negative prognostic impact. In this regard, the aim of our study is to identify clinical severity factors of delirium complicating COVID-19 related-ARDS. We performed a comparative and correlation analysis using demographics, comorbidities, multisystemic and delirium severity scores and anti-delirium therapy in two cohorts of ARDS patients with delirium, respectively, due to COVID-19 ( n = 40) or other medical conditions ( n = 39). Our results indicate that delirium in COVID-19-related ARDS is more severe since its onset despite a relatively less severe systemic condition at the point of ICU admission and required higher dosages of antipsychotic and non-benzodiazepinic sedative therapy respect to non-COVID patients. Finally, the correlation analysis showed a direct association between the male gender and maximum dosage of anti-delirium medications needed within the COVID-19 group, which was taken as a surrogate of delirium severity. Overall, our results seem to indicate that pathogenetic factors specifically associated to severe COVID-19 are responsible for the high severity of delirium, paving the way for future research focused on the mechanisms of the cognitive alterations associated with COVID-19., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Madonna, Enrico, Ciappolino, Boscutti, Colombo, Turtulici, Cantù, Cereda, Delvecchio, De Falco, Chierichetti, Savioli, Grasselli and Brambilla.)

As fathering research has flourished, a growing body of studies has focused on behavioral and neurobiological mechanisms, respectively associated with caregiving sensitivity and responsiveness to infant stimuli. However, the association between these aspects and the key concept of paternal involvement in childcare (i.e., contribution in infant care in terms of time, availability, and responsibility) has been poorly investigated. The current work aims to systematically review the role of involvement in childcare on both neural activations and sensitive behaviors in fathers by examining (a) how paternal involvement has been measured and (b) whether paternal involvement has been associated with neurobiological activation and behavioral sensitive responses. Inclusion criteria were peer-reviewed quantitative studies, concerning fathers responding to infant stimuli at neurobiological or behavioral level, and including a quantitative measurement of paternal involvement in childcare. A quality rating for each study has been performed based on the measurements adopted to assess paternal involvement. Of 2,529 articles, 27 studies were included. According to our quality rating, 10 out of 27 studies included fairly good-standard measures for measuring paternal involvement, whereas 17 studies used good-standard measures. In addition, 11 studies provided details of paternal involvement in the context of neurobiological responses to infant stimuli, whereas 16 addressed paternal sensitive behaviors. Overall, only 8 studies reported relevant findings about the relationship between paternal involvement and neurobiological responses or sensitive behaviors in fathers. The present study is the first systematically evaluating the scope of paternal involvement in the field of Paternal Brain and fathers' sensitive responsiveness research. When high-standard measures are used, paternal involvement seems to play a significant role in modulating both the hormonal and the neural pathways associated with paternal behaviors. Remarkably, the role of paternal engagement may underpin an adaptive nurturance that is not dependent on pregnancy and childbirth but on caregiving experience. A promising positive link between paternal involvement and behavioral sensitivity may be expected in further studies, which will need to corroborate our conclusion by adopting detailed and appropriate measures assessing paternal involvement. As a future line of research, the inclusion of gay fathers may be beneficial for the field., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Giannotti, Gemignani, Rigo, Venuti and De Falco.)

Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Parents of children with autism spectrum disorder (ASD) face unique challenges, which may affect parenting functioning. However, little is known about gender and cultural variations in parenting stress and styles in these families. The aims of this study were to investigate: (1a) the differences in parenting stress and (1b) social style between Italian and Japanese mothers and fathers of children with ASD; (2) the predictive role of culture, sociodemographic, and child's characteristics on parenting stress; (3) the predictors of the social parenting style, including parenting stress dimensions. The study involved 92 Italians and 89 Japanese parents of school-age children (5-12 years) with ASD who completed the Parenting Stress Index and the Parenting Style Questionnaire. Results revealed that Japanese parents showed higher parenting stress and less engagement in social style than Italians. Across cultures, mothers used more social style than fathers. Being Japanese and having a child with greater ASD severity predicted higher levels of parenting stress. We also found that country, parent's gender, and stress related to the dysfunctional interaction were significant predictors of parenting social style. Our findings highlight the importance of a cross-cultural approach to better understand the experiences and needs of mothers and fathers of children with ASD.

Studies on parental interaction in the context of ASD has mainly focused on mothers, even if fathers and their children seem to form close and supportive relationships that may have unique effects on child development. Given the impact of ASD symptoms on a child's ability to interact with significant others, recent findings strengthen the importance of including caregivers during treatment to guarantee a better adaptation to the child's impairments. Despite this, fathers are scarcely involved, and interventions seem to not be tailored to their interactive characteristics and needs. For this reason, a systematic review was conducted to investigate fathers and children with ASD behaviors during interaction. This review found 12 observational studies that identified social, cognitive, and affective interactive modalities in father-child dyads through three psychology-focused journal databases: PubMed, PsycINFO and Scopus. The significant variation in both sample size and in the measures used to assess dyadic outcomes limits the ability of this work to make robust recommendations for intervention. Despite this, the results revealed characteristic behaviors of this dyad that consequently allow specific targets to be worked on during intervention. In fact, from fathers' individual strengths and weaknesses, it is possible to implement interventions that are complementary with maternal characteristics from the perspective of personalized and optimized treatment.

Five vascular endothelial growth factor receptor (VEGFR) ligands (VEGF-A, -B, -C, -D, and placental growth factor [PlGF]) constitute the VEGF family. VEGF-A binds VEGF receptors 1 and 2 (VEGFR1/2), whereas VEGF-B and PlGF only bind VEGFR1. Although much research has been conducted on VEGFR2 to elucidate its key role in retinal diseases, recent efforts have shown the importance and involvement of VEGFR1 and its family of ligands in angiogenesis, vascular permeability, and microinflammatory cascades within the retina. Expression of VEGFR1 depends on the microenvironment, is differentially regulated under hypoxic and inflammatory conditions, and it has been detected in retinal and choroidal endothelial cells, pericytes, retinal and choroidal mononuclear phagocytes (including microglia), Müller cells, photoreceptor cells, and the retinal pigment epithelium. Whilst the VEGF-A decoy function of VEGFR1 is well established, consequences of its direct signaling are less clear. VEGFR1 activation can affect vascular permeability and induce macrophage and microglia production of proinflammatory and proangiogenic mediators. However the ability of the VEGFR1 ligands (VEGF-A, PlGF, and VEGF-B) to compete against each other for receptor binding and to heterodimerize complicates our understanding of the relative contribution of VEGFR1 signaling alone toward the pathologic processes seen in diabetic retinopathy, retinal vascular occlusions, retinopathy of prematurity, and age-related macular degeneration. Clinically, anti-VEGF drugs have proven transformational in these pathologies and their impact on modulation of VEGFR1 signaling is still an opportunity-rich field for further research., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

The anti-HER2 monoclonal antibody trastuzumab is a key drug for the treatment of HER2-positive gastric cancer (GC); however, its activity is often limited by the onset of resistance and mechanisms of resistance are still poorly understood. Several targeted agents showed synergistic activity by concomitant use with trastuzumab in vitro and are under clinical investigation. The aim of this study was to assess the antitumor activity of duligotuzumab, an anti HER3/EGFR antibody or ipatasertib, an AKT inhibitor, combined with trastuzumab in a panel of HER2-positive human gastric cancer cells (GCC), and the efficacy of such combinations in HER2-resistant cells. We have assessed the efficacy of duligotuzumab or ipatasertib and trastuzumab in combination, analyzing proliferation, migration and apoptosis and downstream intracellular signaling in vitro on human HER2-positive GCC (NCI-N87, OE33, OE19) and in negative HER2 GCC (MKN28). We observed a reduction of proliferation, migration and apoptotic rate in HER2-positive OE33, OE19 and N87 cell lines with the combination of duligotuzumab or ipatasertib plus trastuzumab. In particular, in OE33 and OE19 cell lines, the same combined treatment inhibited the activation of proteins downstream of HER2, HER3, AKT and MAPK pathways. Targeting both HER2 and HER3, or HER2 and AKT, results in an improved antitumor effect on HER2-positive GCC.

Background: Few studies have investigated the characteristics of father language directed to typically developing children (TD), and father speech directed to children with Autism Spectrum Disorders (ASD) is largely under investigated. Considering the importance of involving fathers of children with ASD in research and clinical practice, the main purpose of this study was to investigate paternal speech directed to children with ASD compared to that of fathers of TD children., Methods: To this aim, we coded multiple functional aspects of speech during 10-min naturalistic dyadic play interactions between fathers and their preschool children with ASD (n = 20) and with TD (n = 20)., Results: Results showed that fathers of children with ASD displayed a peculiar child-directed language that seems to reflect the effort to provide enhanced scaffolding and reduced demands while sustaining a challenging social interaction. Specifically, fathers of children with ASD used more descriptions, fewer questions in general but more questions about child internal states. Moreover, fathers adapted aspects of their information-salient speech to the severity of child impairments., Conclusions: Our findings support the importance to include fathers in early developmental intervention programs for children with ASD, by underlying fathers' spontaneous adaptation to their children's needs., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Eye drop formulations allowing topical treatment of retinal pathologies have long been sought as alternatives to intravitreal administration. This study aimed to assess whether a novel nanostructured microemulsions system (NaMESys) could be usefully employed to deliver sorafenib to the retina following topical instillation. NaMESys carrying 0.3% sorafenib (NaMESys-SOR) proved to be cytocompatible in vitro on rabbit corneal cells, and well-tolerated following b.i.d. ocular administration to rabbits during a 3-month study. In rats subject to retinal ischemia-reperfusion, NaMESys-SOR significantly inhibited retinal expression of tumor necrosis factor-alpha (TNFα, 20.7%) and inducible nitric oxide synthase (iNos, 87.3%) mRNAs in comparison to controls. Similarly, in streptozotocin-induced diabetic rats, NaMESys-SOR inhibited retinal expression of nuclear factor kappa B (NFκB), TNFα, insulin like growth factor 1 (IGF1), IGF1 receptor (IGF1R), vascular endothelial growth factor receptor 1 (VEGFR1) and 2 (VEGFR2) mRNAs by three-fold on average compared to controls. Furthermore, a reduction in TNFα, VEGFR1 and VEGFR2 protein expression was observed by western blot. Moreover, in mice subject to laser-induced choroidal neovascularization, NaMESys-SOR significantly inhibited neovascular lesions by 54%. In conclusion, NaMESys-SOR was shown to be a well-tolerated ophthalmic formulation able to deliver effective amounts of sorafenib to the retina, reducing proinflammatory and pro-angiogenic mediators in reliable models of proliferative retinopathies. These findings warrant further investigations on the full therapeutic potential of NaMESys-SOR eye drops, aiming to address unmet needs in the pharmacotherapy of retinal neovascular diseases.

Prolyl 3-hydroxylase 2 ( P3H2 ) catalyzes the post-translational formation of 3-hydroxyproline on collagens, mainly on type IV. Its activity has never been directly associated to angiogenesis. Here, we identified P3H2 gene through a deep-sequencing transcriptome analysis of human umbilical vein endothelial cells (HUVECs) stimulated with vascular endothelial growth factor A (VEGF-A). Differently from many previous studies we carried out the stimulation not on starved HUVECs, but on cells grown to maintain the best condition for their in vitro survival and propagation. We showed that P3H2 is induced by VEGF-A in two primary human endothelial cell lines and that its transcription is modulated by VEGF-A/VEGF receptor 2 (VEGFR-2) signaling pathway through p38 mitogen-activated protein kinase (MAPK). Then, we demonstrated that P3H2 , through its activity on type IV Collagen, is essential for angiogenesis properties of endothelial cells in vitro by performing experiments of gain- and loss-of-function. Immunofluorescence studies showed that the overexpression of P3H2 induced a more condensed status of Collagen IV, accompanied by an alignment of the cells along the Collagen IV bundles, so towards an evident pro-angiogenic status. Finally, we found that P3H2 knockdown prevents pathological angiogenesis in vivo, in the model of laser-induced choroid neovascularization. Together these findings reveal that P3H2 is a new molecular player involved in new vessels formation and could be considered as a potential target for anti-angiogenesis therapy.

Research on the psychological impact of the COVID-19 pandemic highlighted negative effects on the general population and particularly on parents. However, little is known about families of children with Neurodevelopmental Disorder (NDD). The present study investigated parental stress, coparenting, and child adjustment in Italian families with children with NDD (N = 82) and typical developing children (TD, N = 82) during lockdown, using an online survey. Results of quantitative analyses showed a significant increase in parental stress and child externalizing behaviors, but not of coparenting. Parental stress is predicted by externalizing behaviors, and coparenting acted as a moderator in the relationship between the change in the amount of time spent with the children before and during lockdown and parental stress. In children with NDD, the decrease in therapeutic/rehabilitation support predicted higher externalizing behaviors. Qualitative analysis showed that beyond the difficulties and worries arising during lockdown, most of the parents appreciated the opportunity of spending more time with their children and strengthening the parent-child relationship. In conclusion, our results point out the importance of ensuring continuity of care for children with NDD (e.g. telehealth) during home confinement and of providing psychological support for parents., (Copyright © 2020. Published by Elsevier Ltd.)

To investigate the role of placental growth factor/vascular endothelial growth factor (PlGF-VEGF) heterodimers are involved in the blood-retinal barrier (BRB) breakdown and the associated mechanism, human retinal endothelial cells (HRECs) were treated with recombinant human (rh)PlGF-VEGF heterodimers and rhPlGF and studied in normal and high-glucose conditions. HREC barrier function was evaluated by the measurement of trans-endothelial electrical resistance (TEER). Adeno-Associated Virus Type 5 (AAV5) vectors overexpressed PlGF in the retina by intravitreal injection into the C57BL6 mouse eye. AAV5-GFP vector and naïve animals were used as controls. Immunofluorescence (IF) and western blots examined the protein expression of PlGF-VEGF heterodimers, VEGF, PlGF, NFκB, p-IκBα, ZO-1, and VE-cadherin in HREC and mouse retina. PlGF-VEGF heterodimers were detected predominantly in the HREC cell nuclei based on IF and cytoplasmic and nuclear fractionation experiments. High glucose treatment increased PlGF-VEGF nuclear abundance. Dot immunoblotting demonstrated a strong affinity of the 5D11D4 antibody to PlGF-VEGF heterodimers. rhPlGF-VEGF disrupted the barrier function of HREC, which was prevented by the neutralization of PlGF-VEGF by the 5D11D4 antibody. Stimulation of HRECs with rhPlGF also led to an increase in the nuclear signals for PlGF-VEGF, p-IκBα, and colocalization of NFκB p65 and PlGF-VEGF in the nuclei. The selective IKK2 inhibitor IMD0354 disrupted the nuclear colocalization. Treatment with IMD0354 restored the barrier function of HREC, as indicated by the ZO-1 and VE-cadherin expression. In the mouse retinas, PlGF overexpression by AAV5 vector reduced ZO-1 expression and increased abundance of pIκBα. PIGF/VEGF heterodimers mediate BRB breakdown potentially through the canonical NFκB activation., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Recombinant antibodies fragments in several new formats are routinely investigated and used in diagnostic and therapeutic applications as anti-cancers molecules. New antibody formats are generated to compensate the need for multispecificity and site-specific introduction of fluorescent dyes, cytotoxic payloads or for generating semisynthetic multimeric molecules. Fabs of trastuzumab bearing transglutaminase (MTG) reactive sites were generated by periplasmic expression in E. coli and purified. Multimeric Fabs were generated by either disulfide bridge formation or by using MTG-sensitive peptide linkers. Binding to receptor was assessed by ELISA and SPR methods. Internalization and growth inhibition assays were performed on BT-474 and SKBR3 Her2+ cells. Fabs were successfully produced and dimerized or trimerized using MTG and suitably designed peptide linkers. Site-specific derivatizations with fluorophores were similarly achieved. The monomeric, dimeric and trimeric variants bind the receptor with affinities similar or superior to the full antibody. Fab and Fab2 are rapidly internalized in Her2+ cells and exhibit growth inhibition abilities similar to the full antibody. Altogether, the data show that the recombinant Fabs can be produced in E. coli and converted into multimeric variants by MTG-based bioconjugation. Similar approaches are extendable to the introduction of cytotoxic payloads for the generation of novel Antibody Drug Conjugates., Competing Interests: Declaration of competing interest The authors declare no conflicts of interests., (Copyright © 2020 Elsevier B.V. All rights reserved.)

(1) Background: Parental involvement during intervention with children with Autism Spectrum Disorder (ASD) has been demonstrated to be fundamental for children's developmental outcomes. However, most research focused on child gains especially considering cognitive functioning and symptoms severity, whereas parental and dyadic changes during intervention need further investigation. (2) Methods: 29 mothers in interaction with their preschool children with ASD were analyzed through two standardized behavioral and observational measures to evaluate the dyadic Emotional Availability (EA) and play skills before (T1) and after (T2) a parental-based intervention. (3) Results: Results revealed mothers increased affective quality and major awareness in understanding the signals produced by the child, that in turn was more responsive, involving also using more complex play strategies. Interestingly, the role of specific factors able to predict parental characteristics was investigated, pointing out the important contribution of mothers' perceptions of having a difficult child and child language communicative abilities. (4) Conclusions: the study enhances knowledge about child and caregiver variables that impact on dyadic outcomes, identifying important target areas to be addressed during intervention. Further, our results suggest that a parental-based intervention supports and facilitates improvements in both children's and caregivers' affective quality and cognitive abilities.

Neural stem cell populations generate a wide spectrum of neuronal and glial cell types in a highly ordered fashion. MicroRNAs are essential regulators of this process. T-UCstem1 is a long non-coding RNA containing an ultraconserved element, and in vitro analyses in pluripotent stem cells provided evidence that it regulates the balance between proliferation and differentiation. Here we investigate the in vivo function of T-UCstem1. We show that T-UCstem1 is expressed in the forebrain neurogenic lineage that generates interneurons for the postnatal olfactory bulb. Gain of function in neural stem cells increased progenitor proliferation at the expense of neuron production, whereas knockdown had the opposite effect. This regulatory function is mediated by its interaction with miR-9-3p and miR-9-5p. Based thereon, we propose a mechanistic model for the role of T-UCstem1 in the dynamic regulation of neural progenitor proliferation during neurogenesis., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: The coronavirus disease 2019 (COVID-19) pandemic has overwhelmed the health systems worldwide. Data regarding the impact of COVID-19 on cancer patients (CPs) undergoing or candidate for immune checkpoint inhibitors (ICIs) are lacking. We depicted the practice and adaptations in the management of patients with solid tumors eligible or receiving ICIs during the COVID-19 pandemic, with a special focus on Campania region., Methods: This survey (25 questions), promoted by the young section of SCITO (Società Campana di ImmunoTerapia Oncologica) Group, was circulated among Italian young oncologists practicing in regions variously affected by the pandemic: high (group 1), medium (group 2) and low (group 3) prevalence of SARS-CoV-2-positive patients. For Campania region, the physician responders were split into those working in cancer centers (CC), university hospitals (UH) and general hospitals (GH). Percentages of agreement, among High (H) versus Medium (M) and versus Low (L) group for Italy and among CC, UH and GH for Campania region, were compared by using Fisher's exact tests for dichotomous answers and χ 2 test for trends relative to the questions with 3 or more options., Results: This is the first Italian study to investigate the COVID-19 impact on cancer immunotherapy, unique in its type and very clear in the results. The COVID-19 pandemic seemed not to affect the standard practice in the prescription and delivery of ICIs in Italy. Telemedicine was widely used. There was high consensus to interrupt immunotherapy in SARS-CoV-2-positive patients and to adopt ICIs with longer schedule interval. The majority of the responders tended not to delay the start of ICIs; there were no changes in supportive treatments, but some of the physicians opted for delaying surgeries (if part of patients' planned treatment approach). The results from responders in Campania did not differ significantly from the national ones., Conclusion: Our study highlights the efforts of Italian oncologists to maintain high standards of care for CPs treated with ICIs, regardless the regional prevalence of COVID-19, suggesting the adoption of similar solutions. Research on patients treated with ICIs and experiencing COVID-19 will clarify the safety profile to continue the treatments, thus informing on the most appropriate clinical conducts., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

The presence of α-synuclein aggregates in the retina of Parkinson's disease patients has been associated with vision impairment. In this study we sought to determine the effects of α-synuclein overexpression on the survival and function of dopaminergic amacrine cells (DACs) in the retina. Adult mice were intravitreally injected with an adeno-associated viral (AAV) vector to overexpress human wild-type α-synuclein in the inner retina. Before and after systemic injections of levodopa (L-DOPA), retinal responses and visual acuity-driven behavior were measured by electroretinography (ERG) and a water maze task, respectively. Amacrine cells and ganglion cells were counted at different time points after the injection. α-synuclein overexpression led to an early loss of DACs associated with a decrease of light-adapted ERG responses and visual acuity that could be rescued by systemic injections of L-DOPA. The data show that α-synuclein overexpression affects dopamine neurons in the retina. The approach provides a novel accessible method to model the underlying mechanisms implicated in the pathogenesis of synucleinopathies and for testing novel treatments.

Alexithymia is defined as a limited ability in the cognitive processing of emotions. Literature suggested its negative influence on interpersonal relationship, documenting elevated alexithymia in individuals with Autism Spectrum Disorder (ASD) compared to control groups. However, the study of alexithymia in school-age children with ASD remains largely unexplored as well as its effect on specific child socioemotional outcomes such as quality of attachment relationships. This study examines alexithymia and perceived attachment to parents in twenty-four children with ASD (without intellectual disability) and 24 typically developing (TD) children (mean age 10 years) using the self-reported Alexithymia Questionnaire for Children (AQC) and the Inventory of Parent and Peer Attachment (IPPA). Measures of family SES as well as child intelligence were collected. Data revealed that ASD children showed higher levels of Alexithymia compared to TD group. In addition, 21% of participants with ASD exceed alexithymia categorical cut-off. By contrast, no difference emerged in the perception of attachment to parents. Moreover, alexithymia, but not ASD status, was found to predictive of child perception of attachment to parents. We observed no significant effect of child age and verbal IQ. Our findings showed that alexithymia was more common in children with ASD, whereas attachment was similar between groups. Difficulties in identifying and describing one's own feelings may hinder the construction of a positive representation of parent-child attachment relationship regardless of child clinical status. Thus, alexithymia seems to play a key role on the way school-age children with and without ASD perceive their relationship with their parents., (Copyright © 2020 Giannotti, de Falco and Venuti.)

Age-related macular degeneration (AMD) is the primary cause of blindness in advanced countries. Repeated intravitreal delivery of anti-vascular endothelial growth factor (VEGF) agents has represented an important advancement for the therapy of wet AMD with significative results in terms of blindness prevention and partial vision restore. Nonetheless, some patients are not responsive or do not attain significant visual improvement, intravitreal injection may cause serious complications and important side effects have been reported for the prolonged block of VEGF-A. In order to evaluate new anti-angiogenic strategies, we focused our attention on VEGF receptor 1 (VEGFR1) developing a specific VEGFR-1 antagonist, a tetrameric tripeptide named inhibitor of VEGFR 1 (iVR1). We have evaluated its anti-angiogenic activity in the preclinical model of AMD, the laser-induced choroid neovascularization (CNV). iVR1 is able to potently inhibit CNV when delivered by intravitreal injection. Surprisingly, it is able to significantly reduce CNV also when delivered by gavage. Our data show that the specific block of VEGFR1 in vivo represents a valid alternative to the block of VEGF-A and that the inhibition of the pathological neovascularization at ocular level is also possible by systemic delivery of compounds not targeting VEGF-A.

Combinatorially generated molecular repertoires have been largely used to identify novel bioactive compounds. Ever more sophisticated technological solutions have been proposed to simplify and speed up such process, expanding the chemical diversity space and increasing the prospect to select new molecular entities with specific and potent activities against targets of therapeutic relevance. In this context, random mixtures of oligomeric peptides were originally used and since 25 years they represent a continuous source of bioactive molecules with potencies ranging from the sub-nM to microM concentration. Synthetic peptide libraries are still employed as starting "synthetic broths" of structurally and chemically diversified molecular fragments from which lead compounds can be extracted and further modified. Thousands of studies have been reported describing the application of combinatorial mixtures of synthetic peptides with different complexity and engrafted on diverse structural scaffolds for the identification of new compounds which have been further developed and also tested in in vivo models of relevant diseases. We briefly review some of the most used methodologies for library preparation and screening and the most recent case studies appeared in the literature where compounds have reached at least in vivo testing in animal or similar models. Recent technological advancements in biotechnology, engineering and computer science have suggested new options to facilitate the discovery of new bioactive peptides. In this instance, we anticipate here a new approach for the design of simple but focused tripeptide libraries against druggable cavities of therapeutic targets and its complementation with existing approaches., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

The impact of different parenting-related variables on child psychological development is widely acknowledged. However, studies about the specific influence of maternal and family dimensions on child early developmental outcomes in at-risk dyads are still scarce. The aim of this longitudinal study was to investigate the short- and middle-term effects of prenatal and postnatal family and maternal features, and child attachment, on child psychological development at 3 and 24 months in at-risk families. Forty-two mothers with psychological, social and/or demographic risk conditions and their first-born infants were assessed longitudinally. Measurements of maternal personality, psychological and depressive symptoms, family socioeconomic status (SES), child-mother attachment, and infant general psychological development were collected at multiple time points, through validated questionnaires and/or mother-child observation. Maternal and family dimensions showed a significant effect on child psychological development over time. The expected detrimental role of reported maternal depressive symptoms was observed both at 3 and 24 months of child's age. Data also highlighted the negative contribution of low family SES and an unexpected positive influence of maternal personality trait of psychoticism on child psychological development at 24 months. We also found a positive association between attachment security and child developmental outcome. These findings might have relevant implications for the implementation of early prevention programs by differentiating the specific predictive role of maternal child and familial factors on child psychological development in at-risk families., (© 2019 Wiley Periodicals, Inc.)

Background: In ICU patients, digestive tract colonization by multidrug-resistant (MDR) Gram-negative (G-) bacteria is a significant risk factor for the development of infections. In patients undergoing extracorporeal membrane oxygenation (ECMO), colonization by MDR bacteria and risk of subsequent nosocomial infections (NIs) have not been studied yet. The aim of this study is to evaluate the incidence, etiology, risk factors, impact on outcome of gastrointestinal colonization by MDR G- bacteria, and risk of subsequent infections in patients undergoing ECMO., Methods: This is a retrospective analysis of prospectively collected data: 105 consecutive patients, treated with ECMO, were admitted to the ICU of an Italian tertiary referral center (San Gerardo Hospital, Monza, Italy) from January 2010 to November 2015. Rectal swabs for MDR G- bacteria were cultured at admission and twice a week. Only colonization and NIs by MDR G- bacteria were analyzed., Results: Ninety-one included patients [48.5 (37-56) years old, 63% male, simplified acute physiology score II 37 (32-47)] underwent peripheral ECMO (87% veno-venous) for medical indications (79% ARDS). Nineteen (21%) patients were colonized by MDR G- bacteria. Male gender (OR 4.03, p = 0.029) and duration of mechanical ventilation (MV) before ECMO > 3 days (OR 3.57, p = 0.014) were associated with increased risk of colonization. Colonized patients had increased odds of infections by the colonizing germs (84% vs. 29%, p < 0.001, OR 12.9), longer ICU length of stay (LOS) (43 vs. 24 days, p = 0.002), MV (50 vs. 22 days, p < 0.001) and ECMO (28 vs. 12 days, p < 0.001), but did not have higher risk of death (survival rate 58% vs. 67%, p = 0.480, OR 0.68). Infected patients had almost halved ICU survival (46% vs. 78%, p < 0.001, OR 4.11)., Conclusions: In patients undergoing ECMO for respiratory and/or circulatory failure, colonization by MDR G- bacteria is frequent and associated with more the tenfold odds for subsequent infection. Those infections are associated with an increased risk of death.

Ex-situ machine perfusion (MP) techniques are increasingly used in clinical settings, especially on grafts derived from donors after cardiac death (DCD). However, comprehension of biological effects elicited during MP are largely unknown and a substantial number of animal studies are presently focused on this topic. The aim of the present study was to describe a model of DCD based on ex-situ perfusion of liver grafts derived from animals dedicated to food production. Procurement took place within a slaughterhouse facility. A clinically fashioned closed circuit normothermic MP (NMP) was built up. Autologous blood-enriched perfusion fluid was adopted. Perfusate and tissue samples were collected to asses NMP functionality. Grafts were classified as transplantable (LT-G) or not (n-LT) according to clinical criteria, while histopathological analysis was used to confirm graft viability. After cold storage, the liver grafts were connected to the NMP. During the rewarming phase, temperature and flows were progressively increased to reach target values. At the end of NMP, 4 grafts were classified as LT-G and 3 nLT-G. Histology confirmed absence of major damage in LT-G, while diffuse necrosis appeared in nLT-G. Interestingly, in nLT-G an early impairment of hepatocyte respiratory chain, leading to cell necrosis and graft non-viability, was documented for the first time. These parameters, together with indocyanine-green dye and citrate clearance could contribute to graft evaluation in clinical settings. In conclusion, this model provides a promising and reproducible method to replace dedicated experimental animals in DCD and MP research, in line with the 3Rs principles.

Diabetic retinopathy (DR) is a secondary complication of diabetes. DR can cause irreversible blindness, and its pathogenesis is considered multifactorial. DR can progress from non-proliferative DR to proliferative DR, characterized by retinal neovascularization. The main cause of vision loss in diabetic patients is diabetic macular edema, caused by vessel leakage and blood retinal barrier breakdown. Currently, aflibercept is an anti-VEGF approved for diabetic macular edema. Aflibercept can bind several members of vascular permeability factors, namely VEGF-A, B, and PlGF. We analyzed the aflibercept-PlGF complex at molecular level, through an in silico approach. In order to explore the role of PlGF in DR, we treated primary human retinal endothelial cells (HRECs) and mouse retinal epithelial cells (RPEs) with aflibercept and an anti-PlGF antibody. We explored the hypothesis that aflibercept has anti-inflammatory action through blocking of PlGF signaling and the ERK axis in an in vitro and in vivo model of DR. Both aflibercept and the anti-PlGF antibody exerted protective effects on retinal cells, by inhibition of the ERK pathway. Moreover, aflibercept significantly decreased (p < 0.05) the expression of TNF-α in an in vitro and in vivo model of DR. Therefore, our data suggest that inhibition of PlGF signaling, or a selective blocking, may be useful in the management of early phases of DR when the inflammatory process is largely involved., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Helper T cells actively communicate with adjacent cells by secreting soluble mediators, yet crosstalk between helper T cells and endothelial cells remains poorly understood. Here we found that placental growth factor (PlGF), a homolog of the vascular endothelial growth factor that enhances an angiogenic switch in disease, was selectively secreted by the T H 17 subset of helper T cells and promoted angiogenesis. Interestingly, the 'angio-lymphokine' PlGF, in turn, specifically induced the differentiation of pathogenic T H 17 cells by activating the transcription factor STAT3 via binding to its receptors and replaced the activity of interleukin-6 in the production of interleukin-17, whereas it suppressed the generation of regulatory T cells. Moreover, T cell-derived PlGF was required for the progression of autoimmune diseases associated with T H 17 differentiation, including experimental autoimmune encephalomyelitis and collagen-induced arthritis, in mice. Collectively, our findings provide insights into the PlGF-dictated links among angiogenesis, T H 17 cell development and autoimmunity.

Background: Normal levels of plasma antithrombin (AT) activity might decrease heparin requirements to achieve an adequate level of anticoagulation during treatment with extracorporeal membrane oxygenation (ECMO). Acquired AT deficiency during ECMO is common, but formal recommendations on target, timing, and rate of AT supplementation are lacking. Thus, we conceived a pilot trial to evaluate the feasibility and safety of prolonged AT supplementation in patients requiring veno-venous ECMO for respiratory failure., Methods: Grifols Antithrombin Research Awards (GATRA) is a prospective, randomized, single blinded, multicenter, controlled two-arm trial. Patients undergoing veno-venous ECMO will be randomized to either receive AT supplementation to maintain a functional AT level between 80 and 120% (AT supplementation group) or not (control group) for the entire ECMO course. In both study groups, anticoagulation will be provided with unfractionated heparin following a standardized protocol. The primary endpoint will be the dose of heparin required to maintain the ratio of activated partial thromboplastin time between 1.5 and 2. Secondary endpoints will be the adequacy of anticoagulation and the incidence of hemorrhagic and thrombotic complications., Discussion: GATRA is a pilot trial that will test the efficacy of a protocol of AT supplementation in decreasing the heparin dose and improving anticoagulation adequacy during ECMO. If positive, it might provide the basis for a future larger trial aimed at verifying the impact of AT supplementation on a composite outcome endpoint including hemorrhagic events, transfusion requirements, and mortality., Trial Registration: ClinicalTrials.gov, NCT03208270 . Registered on 5 July 2017.

The proportion of patients with metastatic colorectal cancer (mCRC) receiving second or further lines of treatment has not been widely studied. To shed light on this issue, we retrospectively analysed the treatments administered for metastatic disease, and investigated prognostic factors after a diagnosis of metastases, in a consecutive cohort of mCRC patients. Three hundred forty-six mCRC patients were enrolled: 173 were stage II or III (metachronous group), and 173 stage IV (synchronous group) at diagnosis. Survival was calculated between the date of metastatic disease and the date of death or last follow-up. Patients with synchronous lesions more frequently had multiple disease sites, peritoneal carcinomatosis and massive liver deposits, whereas significantly more patients with metachronous lesions developed lung metastases as the sole disease site. 97.4% patients received at least one, 62.4% two, 41.9% three and 23.7% four treatment lines. Patients with metachronous metastases more frequently underwent surgery of metastases in first-line treatment (48.5 versus 24.8%), and more of them were progression-free at the time of the analysis (44 versus 34.9%). At univariate analysis, age > 70 years, multiple disease sites and peritoneal carcinomatosis were associated with significantly decreased survival, whereas surgery of metastases and isolated lung metastases predicted better survival. At multivariate analysis, only peritoneal carcinomatosis and surgery of metastases independently affected survival. The percentage of patients who received an active treatment decreased going from first- to fourth-line treatment. However, the proportion of patients who received efficacious treatment in advanced line remained high. Surgery of metastases was the most important prognostic factors.

This study (a) investigates effects of the transition to motherhood on implicit and explicit responses to infant cues; (b) assesses influences of prior parenting and delivery experiences on implicit and explicit responses to infant cues; and (c) investigates relations between implicit and explicit responses to infant cues and parenting beliefs. A total of 45 pregnant women were followed from the sixth month of pregnancy to the third month after the childbirth and were administered a Single Category Implicit Association Test, a semantic differential scale, the Adult Parental Acceptance-Rejection scale, and the Parental Style Questionnaire. The transition to motherhood influenced explicit not implicit responses; only implicit responses were shaped by prior parenting experiences and mode of delivery; and parenting beliefs were related in independent and different ways to implicit and explicit evaluations. These findings indicate that implicit responses are valid and meaningful indices of maternal responsiveness to infants., (© 2018 Wiley Periodicals, Inc.)

Placental growth factor (PlGF) is a proangiogenic member of the vascular endothelial growth factor (VEGF) family playing a central role in pathological angiogenesis. PlGF-DE is a PlGF variant unable to bind vascular endothelial growth factor receptor 1 (VEGFR-1) but still able to generate heterodimer with VEGF-A. We have generated PlGF-DE knockin mice that are vital and fertile and show unaltered expression of Plgf, Vegf-a, Vegfr-1, and Vegfr-2 compared with wild-type mice. Interestingly, these mutants showed additional and remarkable angiogenesis impairment in tumor growth, hindlimb ischemia, and choroidal neovascularization compared with both PlGF knockout and wild-type mice. These findings provided insights on VEGF-A/PlGF heterodimer function, which was able to rescue neovascularization and vascular leakage in PlGF-DE knockin mice. Collectively, these data show that PlGF-DE knockin mouse could be considered the full functional knockout of PlGF, suggesting a reassessment of the phenotypes of knockout mice for the genes whose products are able to generate heterodimeric proteins., (Copyright © 2018 The Author(s). Published by Elsevier Inc. All rights reserved.)

Epigenetic alterations have been associated with both pathogenesis and progression of cancer. By screening of library compounds, we identified a novel hybrid epi-drug MC2884, a HAT/EZH2 inhibitor, able to induce bona fide cancer-selective cell death in both solid and hematological cancers in vitro , ex vivo and in vivo xenograft models. Anticancer action was due to an epigenome modulation by H3K27me3, H3K27ac, H3K9/14ac decrease, and to caspase-dependent apoptosis induction. MC2884 triggered mitochondrial pathway apoptosis by up-regulation of cleaved-BID, and strong down-regulation of BCL2. Even aggressive models of cancer, such as p53 -/- or TET2 -/- cells, responded to MC2884, suggesting MC2884 therapeutic potential also for the therapy of TP53 or TET2-deficient human cancers. MC2884 induced massive apoptosis in ex vivo human primary leukemia blasts with poor prognosis in vivo , by targeting BCL2 expression. MC2884-treatment reduced acetylation of the BCL2 promoter at higher level than combined p300 and EZH2 inhibition. This suggests a key role for BCL-2 reduction in potentiating responsiveness, also in combination therapy with BCL2 inhibitors. Finally, we identified both the mechanism of MC2884 action as well as a potential therapeutic scheme of its use. Altogether, this provides proof of concept for the use of epi-drugs coupled with epigenome analyses to 'personalize' precision medicine., Competing Interests: CONFLICTS OF INTEREST The authors declare that they have no conflict of interest. A patent application that includes some results is being filled with n° WO2017/198870 A1.