A three-gene signature marks the time to locoregional recurrence in luminal-like breast cancer.

BACKGROUND: Gene expression profiling (GEP)-based prognostic signatures are being rapidly integrated into clinical decision making for systemic management of breast cancer patients. However, GEP remains relatively underdeveloped for locoregional risk assessment. Yet, locoregional recurrence (LRR), especially early after surgery, is associated with poor survival. PATIENTS AND METHODS: GEP was carried out on two independent luminal-like breast cancer cohorts of patients developing early (≤5 years after surgery) or late (>5 years) LRR and used, by a training and testing approach, to build a gene signature able to intercept women at risk of developing early LRR. The GEP data of two in silico datasets and of a third independent cohort were used to explore its prognostic value. RESULTS: Analysis of the first two cohorts led to the identification of three genes, CSTB, CCDC91 and ITGB1, whose expression, derived by principal component analysis, generated a three-gene signature significantly associated with early LRR in both cohorts (P value <0.001 and 0.005, respectively), overcoming the discriminatory capability of age, hormone receptor status and therapy. Remarkably, the integration of the signature with these clinical variables led to an area under the curve of 0.878 [95% confidence interval (CI) 0.810-0.945]. In in silico datasets we found that the three-gene signature retained its association, showing higher values in the early relapsed patients. Moreover, in the third additional cohort, the signature significantly associated with relapse-free survival (hazard ratio 1.56, 95% CI 1.04-2.35). CONCLUSIONS: Our three-gene signature represents a new exploitable tool to aid treatment choice in patients with luminal-like breast cancer at risk of developing early recurrence.

Direct quantification of cytosolic delivery of drug nanocarriers using FlAsH-EDT2.

The delivery of therapeutics across the cell membrane and into the cytoplasm is a major challenge that limits the development of new therapies. This challenge is compounded by the lack of a general assay for cytosolic delivery. Here we develop this assay based on the pro-fluorophore CrAsH-EDT2, and provide cytosolic penetration results for a variety of drug delivery agents (polyethyleneimine, poly-arginine, Ferritin, poly [maleic anhydride-alt-isobutene] grafted with dodecylamine, and cationic liposomes) into HeLa and T98G cells. Our results show that this method can be widely applicable to different cells and drug delivery agents, and yield statistically robust results. We later use this method to optimize and improve a model drug delivery agent's (Ferritin) cytosolic penetration.

Motor and Cognitive Modulation of a Single Session of Transcutaneous Auricular Vagus Nerve Stimulation in Post Stroke Patients: A Pilot Study.

Objective: The aim of the present study is to explore whether a single session of transcutaneous Vagus Nerve Stimulation (tVNS) can enhance the ipsilesional, and contralesional upper limb motor functions as well as cognitive functions in stroke patients. The effects of the stimulation were evaluated through two different tasks: the box and blocks test (BB), indexing manual dexterity, and the Go/No-go task, a visuomotor paradigm used to assess both motor readiness and response inhibition. Tests were administered without tVNS, during tVNS and during sham tVNS. Results: The BB showed a statistical difference for both contralesional side (p = 0.05) between Basal-Real condition (p = 0.042) and ipsilesional side (p = 0.001) between Basal-Real (p = 0.008) and for Real-Sham (p = 0.005). Any statistical difference was found for the mean latencies in the three conditions of the Go/No-go test. Conclusion: A single session of tVNS seems to improve upper limb motor functions but not cognitive functions in post-stroke patients, despite a positive trend was detected.

PerBrain: a multimodal approach to personalized tracking of evolving state-of-consciousness in brain-injured patients: protocol of an international, multicentric, observational study.

BACKGROUND: Disorders of consciousness (DoC) are severe neurological conditions in which consciousness is impaired to various degrees. They are caused by injury or malfunction of neural systems regulating arousal and awareness. Over the last decades, major efforts in improving and individualizing diagnostic and prognostic accuracy for patients affected by DoC have been made, mainly focusing on introducing multimodal assessments to complement behavioral examination. The present EU-funded multicentric research project "PerBrain" is aimed at developing an individualized diagnostic hierarchical pathway guided by both behavior and multimodal neurodiagnostics for DoC patients. METHODS: In this project, each enrolled patient undergoes repetitive behavioral, clinical, and neurodiagnostic assessments according to a patient-tailored multi-layer workflow. Multimodal diagnostic acquisitions using state-of-the-art techniques at different stages of the patients' clinical evolution are performed. The techniques applied comprise well-established behavioral scales, innovative neurophysiological techniques (such as quantitative electroencephalography and transcranial magnetic stimulation combined with electroencephalography), structural and resting-state functional magnetic resonance imaging, and measurements of physiological activity (i.e. nasal airflow respiration). In addition, the well-being and treatment decision attitudes of patients' informal caregivers (primarily family members) are investigated. Patient and caregiver assessments are performed at multiple time points within one year after acquired brain injury, starting at the acute disease phase. DISCUSSION: Accurate classification and outcome prediction of DoC are of crucial importance for affected patients as well as their caregivers, as individual rehabilitation strategies and treatment decisions are critically dependent on the latter. The PerBrain project aims at optimizing individual DoC diagnosis and accuracy of outcome prediction by integrating data from the suggested multimodal examination methods into a personalized hierarchical diagnosis and prognosis procedure. Using the parallel tracking of both patients' neurological status and their caregivers' mental situation, well-being, and treatment decision attitudes from the acute to the chronic phase of the disease and across different countries, this project aims at significantly contributing to the current clinical routine of DoC patients and their family members. TRIAL REGISTRATION: ClinicalTrials.gov, NCT04798456 . Registered 15 March 2021 - Retrospectively registered.

Common practices in patient care after mammaplasty and perioperative safety measures to prevent thromboembolic events - An observational study across all continents.

BACKGROUND: Most plastic surgeons have encountered complications related to venous thromboembolism in their profession. However, considerable deficits in risk assessment and prophylaxis have been described. As outpatient procedures are increasing, so are concerns with regard to patient safety. The presented study investigates the global distribution pattern of patient care for mammaplasties and compares international common practices of perioperative safety measures to prevent thromboembolic events. METHODS: A questionnaire was sent to over 5000 plastic surgeons in 77 countries. The survey inquired about surgeon demographics, standard perioperative safety measures, type of thrombosis prophylaxis, and international patterns of patient care. The results were evaluated and correlated with the evidence-based literature. RESULTS: A total of 1431 surveys were gathered (response rate: 29%). The gathered data show international disparity with regard to inpatient vs. outpatient care. Mammaplasties in the USA are being performed as same-day surgeries in > 80% of cases, while globally inpatient and outpatient procedures are distributed evenly. Also, we found no international consensus with regard to safety measures to prevent thromboembolic events. Geographical regions showed differences with regard to type, dosage, timing, and duration of thrombosis prophylaxis. CONCLUSIONS: International practice patterns remain incoherent. This highlights the need for coherent and detailed global guidelines for plastic surgical interventions. High-quality studies are needed in order to establish evidence-based, standardized, and universally applicable practice guidelines.

Guía para la práctica de acogimiento familiar, relato de una experiencia / Guidelines for being a foster care home, report of an experience

Recibir a un niño o a una niña en cuidado transitorio supone la fortaleza para enfrentar situaciones complejas, ofrecer cuidados y contención, y ser consciente del carácter temporario de ese hecho. El pediatra juega un papel determinante no solo en el cuidado de la salud de niños, niñas y adolescentes con derechos vulnerados, sino también como actor clave en la detección temprana de los casos, a partir de una mirada atenta y comprometida con la situación de cada niño. Se presenta un caso de 5 hermanos con derechos vulnerados, en el que intervino el Sistema de Promoción y Protección Integral de Derechos de los Niños a través de un programa de familias de abrigo, con el objetivo de clarificar la situación legal actual y concientizar sobre dicha alternativa.

Receiving a child in foster home care needs the strength to face complex situations, gives care and support, and be aware of the temporary nature of that position. The pediatrician plays a decisive role not only in the health care of children and adolescents with lost rights, but also in the early detection of cases, based on an attentive and committed attitudes facing each child circumstances. We present the experience of 5 siblings with lost rights in which the Promotion and Integral Protection of Children's Rights System intervened through a program of foster families, with the aim of clarifying the current legal situation and raising awareness about this alternative.

Guidelines for being a foster care home, report of an experience. / Guía para la práctica de acogimiento familiar, relato de una experiencia.

Receiving a child in foster home care needs the strength to face complex situations, gives care and support, and be aware of the temporary nature of that position. The pediatrician plays a decisive role not only in the health care of children and adolescents with lost rights, but also in the early detection of cases, based on an attentive and committed attitudes facing each child circumstances. We present the experience of 5 siblings with lost rights in which the Promotion and Integral Protection of Children's Rights System intervened through a program of foster families, with the aim of clarifying the current legal situation and raising awareness about this alternative.

Recibir a un niño o a una niña en cuidado transitorio supone la fortaleza para enfrentar situaciones complejas, ofrecer cuidados y contención, y ser consciente del carácter temporario de ese hecho. El pediatra juega un papel determinante no solo en el cuidado de la salud de niños, niñas y adolescentes con derechos vulnerados, sino también como actor clave en la detección temprana de los casos, a partir de una mirada atenta y comprometida con la situación de cada niño. Se presenta un caso de 5 hermanos con derechos vulnerados, en el que intervino el Sistema de Promoción y Protección Integral de Derechos de los Niños a través de un programa de familias de abrigo, con el objetivo de clarificar la situación legal actual y concientizar sobre dicha alternativa.

EEG spectral exponent as a synthetic index for the longitudinal assessment of stroke recovery.

OBJECTIVE: Quantitative Electroencephalography (qEEG) can capture changes in brain activity following stroke. qEEG metrics traditionally focus on oscillatory activity, however recent findings highlight the importance of aperiodic (power-law) structure in characterizing pathological brain states. We assessed neurophysiological alterations and recovery after mono-hemispheric stroke by means of the Spectral Exponent (SE), a metric that reflects EEG slowing and quantifies the power-law decay of the EEG Power Spectral Density (PSD). METHODS: Eighteen patients (n = 18) with mild to moderate mono-hemispheric Middle Cerebral Artery (MCA) ischaemic stroke were retrospectively enrolled for this study. Patients underwent EEG recording in the sub-acute phase (T0) and after 2 months of physical rehabilitation (T1). Sixteen healthy controls (HC; n = 16) matched by age and sex were enrolled as a normative group. SE values and narrow-band PSD were estimated for each recording. We compared SE and band-power between patients and HC, and between the affected (AH) and unaffected hemisphere (UH) at T0 and T1 in patients. RESULTS: At T0, stroke patients showed significantly more negative SE values than HC (p = 0.003), reflecting broad-band EEG slowing. Most important, in patients SE over the AH was consistently more negative compared to the UH and showed a renormalization at T1. This SE renormalization significantly correlated with National Institute of Health Stroke Scale (NIHSS) improvement (R = 0.63, p = 0.005). CONCLUSIONS: SE is a reliable readout of the neurophysiological and clinical alterations occurring after an ischaemic cortical lesion. SIGNIFICANCE: SE promise to be a robust method to monitor and predict patients' functional outcome.

Development of an integrated environmental monitoring protocol for SARS-CoV-2 contamination. Applications at the IRCSS San Martino Polyclinic Hospital in Genoa, Italy.

SCIENTIFIC BACKGROUND: Environmental sampling of SARS-CoV-2 is a fundamental tool for evaluating the effectiveness of non-specific prophylaxis measures in counteracting virus spread. The purpose of our work was to evaluate the effectiveness of the different sampling methods in the hospital setting to assess their correlation with the structural, functional, and operational situation of the monitored departments and to define the dynamics of the spread of the virus in indoor environments. METHODS: The monitoring (air bubbling sampling, surface wipe test) was carried out at the San Martino Polyclinic Hospital (Genoa, Italy) in the period since April 2020 to June 2021. The presence of viral RNA in the collected samples was evaluated by qPCR. The infection capacity of the samples collected was also evaluated by an in vitro challenge test on cells sensitive to SARS-CoV-2 infection. RESULTS: The percentage of positivity with respect to the number of tests performed (sensitivity) were air bubbler 50%, wipe test 17%, and challenge test 11%. Only 20% of the samples tested positive in the wipe test and 43% of the samples tested positive in the bubbler sampling were also positive in the challenge test. All the positivity obtained was detected at a distance of less than 2 m and height of less than 1.5 from COVID-19 patients. CONCLUSIONS: Environmental contamination from SARS-CoV-2 detected at the San Martino Polyclinic Hospital is found lower than similar assessments performed in other hospitals both in Italy and abroad. Our study predicted that environmental monitoring of SARS-CoV-2 must be carried out in an integrated way by not using a single sampling method, as each individual test has a different biological significance and performance. However, the virus detected by wipe test only is often a degraded viral fragment and not an intact infecting virion.

Long-term clinical outcome of dental implants: A retrospective clinical study with a minimum follow-up between 9.5 and 17.7 years.

The aim of this retrospective case series was to evaluate the clinical and radiographic outcomes of the patients that underwent implant surgery in all indication classes, with a follow-up of at least 9 years. 121 healthy patients in need for oral rehabilitation with dental implants were included in this study. 196 implants (160 conical, 73 cylindric design implants) were inserted. The implant survival rate was the primary outcome. Intra- and postoperative complications were additional criteria for success. The mean follow-up of the patients was 12.29 years (SD 1.39). Mean age of the study population was 51.0 years (SD 12.7). The mean bone loss around implants after at least 9 years of loading was measured as 2.0 mm (SD 0.73 mm). Intra-operative complications were seen in 5 patients. Post-operative complications included: 5 mucositis,1 dehiscence, 2 screw loosening, 1 infection at site and 1 nonintegrated implant. Two implants were lost in two patients. The overall implant survival rate was 99.1%. As a conclusion, oral rehabilitation with dental implant-supported prostheses can be accepted as a safe procedure with relevantly high survival rates of oral implants and successful aesthetic and functional outcomes.

Artichoke cv. Francés flower extract as a rennet substitute: effect on textural, microstructural, microbiological, antioxidant properties, and sensory acceptance of miniature cheeses.

BACKGROUND: The most common milk-clotting enzymes in the cheese industry are recombinant chymosins. Food naturalness is a factor underpinning consumers' food choice. For consumers who avoid food with ingredients from genetically modified organisms (GMOs), the use of vegetable-based rennet substitute in the cheese formulation may be a suitable solution. Artichokes that deviate from optimal products, when allowed to bloom due to flower protease composition, are excellent as raw material for vegetable rennet preparation. As enzymatic milk clotting exerts a significant impact on the characteristics of the final product, this product should be studied carefully. RESULTS: Mature flowers from unharvested artichokes (Cynara scolymus cv. Francés) that did not meet aesthetic standards for commercialization were collected and used to prepare a flower extract. This extract, as a coagulant preparation, enabled the manufacture of cheeses with distinctive characteristics compared with cheeses prepared with chymosin. Rennet substitution did not affect the actual yield but led to significant changes in dry matter yield, humidity, water activity, protein content, and color, and conferred antioxidant activity to the cheeses. The rennet substitution promoted significant modifications in springiness, and in the microstructure of the cheese, with a more porous protein matrix and an increment in the size of the fat globules. Both formulations showed a similar microbiota evolution pattern with excellent microbiological quality and good sensory acceptance. CONCLUSIONS: The rennet substitute studied here produced a cheese adapted to specific market segments that demand more natural and healthier products made with a commitment to the environment but well accepted by a general cheese consumer. © 2020 Society of Chemical Industry.

Immunometabolic Status of COVID-19 Cancer Patients.

Cancer patients appear to be more likely to be diagnosed with coronavirus disease 2019 (COVID-19). This is supported by the understanding of immunometabolic pathways that intersect patients with infection and cancer. However, data derived by case series and retrospective studies do not offer a coherent interpretation, since data from China suggest an increased risk of COVID-19, while data from the United States and Italy show a prevalence of COVID-19 in cancer patients comparable with the general population. Noteworthy, cancer and COVID-19 exploit distinct patterns of macrophage activation that promote disease progression in the most severe forms. In particular, the alternative activation of M2-polarized macrophages plays a crucial role in cancer progression. In contrast, the macrophage-activation syndrome appears as the source of M1-related cytokine storm in severe COVID-19 disease, thus indicating macrophages as a source of distinct inflammatory states in the two diseases, nonetheless as a common therapeutic target. New evidence indicates that NAMPT/NAD metabolism can direct both innate immune cell effector functions and the homeostatic robustness, in both cancer and infection. Moreover, a bidirectional relationship exists between the metabolism of NAD and the protective role that angiotensin converting enzyme 2, the COVID-19 receptor, can play against hyperinflammation. Within this immunometabolic framework, the review considers possible interference mechanisms that viral infections and tumors elicit on therapies and provides an overview for the management of patients with cancer affected by COVID-19, particularly for the balance of risk and benefit when planning normally routine cancer treatments and follow-up appointments.

Claw-in-the-door: pigeons, like humans, display the foot-in-the-door effect.

People are more likely to comply with a large request when it is preceded by another, smaller request, and this is known as the "foot-in-the-door" (FITD). The FITD has been widely studied in social psychology and is thought to arise from mutually conflicting beliefs about past and present behavior (cognitive dissonance) or changes in self-perception. Across two experiments, we found that pigeons' latency to respond to an effortful second stimulus in a pair scales with how much effort they had exerted on the first stimulus. As such, pigeons also display a FITD-like effect. We argue that the FITD may not be caused by conflicting beliefs or changes in self-perception but may instead be the product of behavioral contrast.

VPA/PLGA microfibers produced by coaxial electrospinning for the treatment of central nervous system injury.

The central nervous system shows limited regenerative capacity after injury. Spinal cord injury (SCI) is a devastating traumatic injury resulting in loss of sensory, motor, and autonomic function distal from the level of injury. An appropriate combination of biomaterials and bioactive substances is currently thought to be a promising approach to treat this condition. Systemic administration of valproic acid (VPA) has been previously shown to promote functional recovery in animal models of SCI. In this study, VPA was encapsulated in poly(lactic-co-glycolic acid) (PLGA) microfibers by the coaxial electrospinning technique. Fibers showed continuous and cylindrical morphology, randomly oriented fibers, and compatible morphological and mechanical characteristics for application in SCI. Drug-release analysis indicated a rapid release of VPA during the first day of the in vitro test. The coaxial fibers containing VPA supported adhesion, viability, and proliferation of PC12 cells. In addition, the VPA/PLGA microfibers induced the reduction of PC12 cell viability, as has already been described in the literature. The biomaterials were implanted in rats after SCI. The groups that received the implants did not show increased functional recovery or tissue regeneration compared to the control. These results indicated the cytocompatibility of the VPA/PLGA core-shell microfibers and that it may be a promising approach to treat SCI when combined with other strategies.

VPA/PLGA microfibers produced by coaxial electrospinning for the treatment of central nervous system injury

The central nervous system shows limited regenerative capacity after injury. Spinal cord injury (SCI) is a devastating traumatic injury resulting in loss of sensory, motor, and autonomic function distal from the level of injury. An appropriate combination of biomaterials and bioactive substances is currently thought to be a promising approach to treat this condition. Systemic administration of valproic acid (VPA) has been previously shown to promote functional recovery in animal models of SCI. In this study, VPA was encapsulated in poly(lactic-co-glycolic acid) (PLGA) microfibers by the coaxial electrospinning technique. Fibers showed continuous and cylindrical morphology, randomly oriented fibers, and compatible morphological and mechanical characteristics for application in SCI. Drug-release analysis indicated a rapid release of VPA during the first day of the in vitro test. The coaxial fibers containing VPA supported adhesion, viability, and proliferation of PC12 cells. In addition, the VPA/PLGA microfibers induced the reduction of PC12 cell viability, as has already been described in the literature. The biomaterials were implanted in rats after SCI. The groups that received the implants did not show increased functional recovery or tissue regeneration compared to the control. These results indicated the cytocompatibility of the VPA/PLGA core-shell microfibers and that it may be a promising approach to treat SCI when combined with other strategies.

Ursolic acid: A novel antiviral compound inhibiting rotavirus infection in vitro.

Rotavirus is one of the leading causes of severe acute gastroenteritis in children under 5 years of age, mainly affecting developing countries. Once the disease is acquired, no specific treatment is available; as such, the development of new drugs for effective antirotaviral treatment is critical. Ursolic acid is a pentacyclic triterpenoid with antiviral activity, which has been studied extensively in vitro and in vivo. To study the potential antirotaviral activity of ursolic acid, its toxic potential for viral particles (virucidal effect) and cultured cells (cytotoxicity) was analysed. No effect on virion infectivity was observed with treatments of up to 40 µM ursolic acid, while incipient cytotoxicity started to be evident with 20 µM ursolic acid. The antiviral potential of ursolic acid was evaluated in in-vitro rotavirus infections, demonstrating that 10 µM ursolic acid inhibits rotavirus replication (observed by a decrease in viral titre and the level of the main viral proteins) and affects viral particle maturation (a process associated with the endoplasmic reticulum) 15 h post infection. Interestingly, ursolic acid was also found to hamper the early stages of the viral replication cycle, as a significant reduction in the number and size of viroplasms, consistent with a decrease in VP6 and NSP2 viral proteins, was observed 4 h post infection. As such, these observations demonstrate that ursolic acid exhibits antiviral activity, suggesting that this chemical could be used as a new treatment for rotavirus.

Choosing wisely first line immunotherapy in non-small cell lung cancer (NSCLC): what to add and what to leave out.

Immunotherapy has dramatically changed the therapeutic scenario in treatment naïve advanced non-small cell lung cancer (NSCLC). While single agent pembrolizumab has become the standard therapy in patients with PD-L1 expression on tumor cells ≥ 50%, the combination of pembrolizumab or atezolizumab and platinum-based chemotherapy has emerged as an effective first line treatment regardless of PD-L1 expression both in squamous and non-squamous NSCLC without oncogenic drivers. Furthermore, double immune checkpoint inhibition has shown promising results in treatment naïve patients with high tumor mutational burden (TMB). Of note, the presence of both negative PD-L1 expression and low TMB may identify a subgroup of patients who has little benefit from immunotherapy combinations and for whom the best treatment option may still be platinum-based chemotherapy. To date, first-line single agent immune checkpoint blockade has demonstrated limited activity in EGFR mutated NSCLC and the combination of immunotherapy and targeted agents has raised safety concerns in both EGFR and ALK positive NSCLC patients. Finally, in EGFR mutated or ALK rearranged NSCLC, atezolizumab in combination with platinum-based chemotherapy and bevacizumab is emerging as a potential treatment option upon progression to first line tyrosine kinase inhibitors.

Retrospective, multicenter evaluation of complications in the treatment of diaphyseal femur fractures in pediatric patients.

INTRODUCTION: Femoral shaft fractures are the commonest major pediatric fractures. For generations, traction and casting were the standard method of treatment for children. However, over the past two decades there has been growing recognition of the advantages of fixation and rapid mobilization. METHODS: A prospective multicenter study was conducted at four Italian centers of reference for pediatric fractures (January 2005 to December 2014). The study involved 62 patients of both sexes, between 6 and 14 years of age, with closed femoral shaft fractures. The aim was to find out more about the short-term complications of titanium elastic nailing in diaphyseal femur fractures in children in order to reduce them. RESULTS: The commonest complication observed in our study was pain at the nail entry point (24.19%) due to a local inflammatory reaction. After 1 year, 3.22% had limbs of different lengths. Proximal migration occurred in 1.61% of cases. DISCUSSION: Over the last two decades, the treatment of femoral shaft fractures in pediatric patients has developed to include internal fixation using Titanium Elastic Nails (TEN). We only observed a few complications in our study, most of which were minor and associated with the surgical technique employed, particularly during the initial phase of the surgeon's learning curve. CONCLUSIONS: TEN are an excellent internal fixation system if used by an expert surgeon and have a very low rate of complications. None of them produced permanent damage in the patients. In older children weighing more than 50 kg, alternative techniques such as subtrochanteric nailing, plates, or external fixation are advisable.