Dynamic co-evolution of transposable elements and the piRNA pathway in African cichlid fishes.

East African cichlid fishes have diversified in an explosive fashion, but the (epi)genetic basis of the phenotypic diversity of these fishes remains largely unknown. Although transposable elements (TEs) have been associated with phenotypic variation in cichlids, little is known about their transcriptional activity and epigenetic silencing. Here, we describe dynamic patterns of TE expression in African cichlid gonads and during early development. Orthology inference revealed an expansion of piwil1 genes in Lake Malawi cichlids, likely driven by PiggyBac TEs. The expanded piwil1 copies have signatures of positive selection and retain amino acid residues essential for catalytic activity. Furthermore, the gonads of African cichlids express a Piwi-interacting RNA (piRNA) pathway that target TEs. We define the genomic sites of piRNA production in African cichlids and find divergence in closely related species, in line with fast evolution of piRNA-producing loci. Our findings suggest dynamic co-evolution of TEs and host silencing pathways in the African cichlid radiations. We propose that this co-evolution has contributed to cichlid genomic diversity.

Ultrasound-Guided Resection of High-Grade Gliomas: A Single-Arm Meta-Analysis.

BACKGROUND: High-grade gliomas (HGGs) present a challenge in neuro-oncology, often necessitating surgical resection for optimal management. Ultrasound holds promise in achieving better gross total resection (GTR) and improving outcomes. This meta-analysis systematically evaluates literature providing robust evidence on the use of intraoperative ultrasonography (iUSG) in HGG resection. METHODS: Following Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines a comprehensive search was made across PubMed, Embase, Cochrane, and Web of Science utilized terms related to iUSG for HGG resection. The meta-analysis examined randomized trials and observational cohort studies on iUSG-guided HGG resection. GTR, subtotal resection, and postresection complications were assessed. Statistical analysis, employing R software for a single proportion analysis with confidence intervals of 95%, I2 statistics for heterogeneity, and the instrumental variables method with restricted maximum likelihood for a random effects model. RESULTS: A total of 178 patients were included in our study. The GTR overall rate in patients with iUSG-guided resection was found to be 64% (95% confidence interval: 46%-81%). Two-dimensional ultrasound remains dominant at 80% against other options of ultrasound. Complications were reported at a 15% rate (95% confidence interval: 7%-23%). CONCLUSIONS: Our study provided robust data on the utilization of iUSG-guided resection regarding the attainment of GTR and the complications related to resection. However, challenges such as outcome heterogeneity and limited complication reporting highlight the need for further research to optimize iUSG in HGG treatment. Long-term follow-up studies on patient survival and postsurgery quality of life will complement existing literature, guiding clinical practices in managing HGG.

Primary EGIST of the greater omentum - a rare presentation.

Extragastrointestinal stromal tumors (EGISTs) are rare mesenchymal neoplasms, which develop in the retroperitoneum, mesentery, and omentum, lacking continuity to the stomach or intestines. Authors hereby present a female patient with a large heterogeneous abdominal mass as a case of an omental EGIST. A 46-year-old woman was referred to our hospital due to an insidious enlargement and colicky pain in the right iliac fossa. Abdominal palpation revealed a voluminous, freely mobile, and non-pulsatile mesoabdominal bulge expanding to the hypogastrium. On exploratory midline laparotomy, the tumor was densely fused to the greater omentum, not connected to the stomach, without gross involvement of adjacent structures. The large mass was completely excised after adequate mobilization. Immunohistochemical techniques showed strong and diffuse expression of WT1, actin and DOG-1, as well as multifocal c-KIT marking. Mutational study concluded a double mutation of KIT exon 9 and a mutation of PDGFRA exon 18. The patient was submitted to adjuvant treatment with imatinib mesylate 800 mg/day. Despite an extremely diverse presentation, omental EGISTs often remain clinically silent for a long time having enough space to grow before becoming symptomatic. These tumors have a consistent pattern of metastasis that typically spares lymph nodes unlike epithelial gut neoplasms. Surgery remains the preferred treatment for non-metastatic EGISTs of the greater omentum. It is possible that DOG-1 will supplant KIT as the leading marker in the future. The scarcity of knowledge on omental EGISTs implies a close monitoring of these patients to detect local relapse or distant metastasis.

Genome editing in East African cichlids and tilapias: state-of-the-art and future directions.

African cichlid fishes of the Cichlidae family are a group of teleosts important for aquaculture and research. A thriving research community is particularly interested in the cichlid radiations of the East African Great Lakes. One key goal is to pinpoint genetic variation underlying phenotypic diversification, but the lack of genetic tools has precluded thorough dissection of the genetic basis of relevant traits in cichlids. Genome editing technologies are well established in teleost models like zebrafish and medaka. However, this is not the case for emerging model organisms, such as East African cichlids, where these technologies remain inaccessible to most laboratories, due in part to limited exchange of knowledge and expertise. The Cichlid Science 2022 meeting (Cambridge, UK) hosted for the first time a Genome Editing Workshop, where the community discussed recent advances in genome editing, with an emphasis on CRISPR/Cas9 technologies. Based on the workshop findings and discussions, in this review we define the state-of-the-art of cichlid genome editing, share resources and protocols, and propose new possible avenues to further expand the cichlid genome editing toolkit.

Age- and caste-independent piRNAs in the germline and miRNA profiles linked to caste and fecundity in the ant Temnothorax rugatulus.

Social insects are models for studies of phenotypic plasticity. Ant queens and workers vary in fecundity and lifespan, which are enhanced and extended in queens. Yet, the regulatory mechanisms underlying this variation are not well understood. Ant queens live and reproduce for years, so that they need to protect their germline from transposable element (TE) activity, which may be redundant in short-lived, often sterile workers. We analysed the expression of two protective classes of small RNAs, microRNAs (miRNAs) and Piwi-interacting RNAs (piRNAs), in various tissues, castes and age classes of the ant Temnothorax rugatulus. In queens, piRNAs were highly abundant in ovaries with TEs being their clear targets, with reduced but still detectable piRNA-specific ping-pong signatures in thorax and brains. piRNA pathway activity varied little with age in queens. Moreover, the reduced ovaries of workers also exhibited similar piRNA activity and this not only in young, fertile workers, but also in older foragers with regressed ovaries. Therefore, these ants protect their germline through piRNA activity, regardless of ovarian development, age or caste, even in sterile workers often considered the soma of the superorganism. Our tissue-specific miRNA analysis detected the expression of 304 miRNAs, of which 105 were expressed in all tissues, 10 enriched in the brain, three in the thorax, whereas 83 were ovarian-specific. We identified ovarian miRNAs whose expression was related to caste, fecundity and age, and which likely regulate group-specific gene expression. sRNA shifts in young- to middle-aged queens were minor, suggesting delayed senescence in this reproductive caste.

Urinary continence recovery after Retzius-sparing robot-assisted radical prostatectomy in relation to surgeon experience.

Urinary incontinence is one of the main concerns for patients after radical prostatectomy. Differences in surgical experience among surgeons could partly explain the wide range of frequencies observed. Our aim was to evaluate the association between the surgeons` experience and center caseload with relation to urinary continence recovery after Retzius-sparing robot-assisted radical prostatectomy (RS-RARP). Prospective observational single-center study. Five surgeons consecutively operated 405 patients between July 2017 and February 2022. Continence recovery was evaluated with pad count and by employing the short form of the International Consultation on Incontinence Questionnaire (ICIQ-SF), pre- and postoperatively at 1 year. Non-parametric tests were used. Median age was 63 years, 30% of patients presented with local advanced disease; the positive surgical margin rate (over 3 mm length) was 16%. Complication rate was 1% (Clavien-Dindo > II). One year after surgery, continence was assessed in 282 patients, of whom 87% were pad free and 51% never leaked (ICIQ-SF = 0). With respect to the mean annual number of procedures per surgeon, divided in < 20, 20-39 and ≥ 40, pad-free rates were achieved in 93%, 85%, and 84% and absence of urine leak rates in 47%, 62% and 48% of patients, respectively. Postoperative median ICIQ-SF was five. We acknowledge the limitation of a 12-month follow-up and the fact that we are a medium-volume center. There is no statistically significant association between continence recovery, surgeon's experience and center caseload. Continence recovery at 1 year after surgery is adequate and robust to surgeon's experience.

COVID-19 Contact Tracing as an Indicator for Evaluating a Pandemic Situation: Simulation Study.

BACKGROUND: Contact tracing is a fundamental intervention in public health. When systematically applied, it enables the breaking of chains of transmission, which is important for controlling COVID-19 transmission. In theoretically perfect contact tracing, all new cases should occur among quarantined individuals, and an epidemic should vanish. However, the availability of resources influences the capacity to perform contact tracing. Therefore, it is necessary to estimate its effectiveness threshold. We propose that this effectiveness threshold may be indirectly estimated using the ratio of COVID-19 cases arising from quarantined high-risk contacts, where higher ratios indicate better control and, under a threshold, contact tracing may fail and other restrictions become necessary. OBJECTIVE: This study assessed the ratio of COVID-19 cases in high-risk contacts quarantined through contact tracing and its potential use as an ancillary pandemic control indicator. METHODS: We built a 6-compartment epidemiological model to emulate COVID-19 infection flow according to publicly available data from Portuguese authorities. Our model extended the usual susceptible-exposed-infected-recovered model by adding a compartment Q with individuals in mandated quarantine who could develop infection or return to the susceptible pool and a compartment P with individuals protected from infection because of vaccination. To model infection dynamics, data on SARS-CoV-2 infection risk (IR), time until infection, and vaccine efficacy were collected. Estimation was needed for vaccine data to reflect the timing of inoculation and booster efficacy. In total, 2 simulations were built: one adjusting for the presence and absence of variants or vaccination and another maximizing IR in quarantined individuals. Both simulations were based on a set of 100 unique parameterizations. The daily ratio of infected cases arising from high-risk contacts (q estimate) was calculated. A theoretical effectiveness threshold of contact tracing was defined for 14-day average q estimates based on the classification of COVID-19 daily cases according to the pandemic phases and was compared with the timing of population lockdowns in Portugal. A sensitivity analysis was performed to understand the relationship between different parameter values and the threshold obtained. RESULTS: An inverse relationship was found between the q estimate and daily cases in both simulations (correlations >0.70). The theoretical effectiveness thresholds for both simulations attained an alert phase positive predictive value of >70% and could have anticipated the need for additional measures in at least 4 days for the second and fourth lockdowns. Sensitivity analysis showed that only the IR and booster dose efficacy at inoculation significantly affected the q estimates. CONCLUSIONS: We demonstrated the impact of applying an effectiveness threshold for contact tracing on decision-making. Although only theoretical thresholds could be provided, their relationship with the number of confirmed cases and the prediction of pandemic phases shows the role as an indirect indicator of the efficacy of contact tracing.

Silver Nanostar-Based SERS for the Discrimination of Clinically Relevant Acinetobacter baumannii and Klebsiella pneumoniae Species and Clones.

The development of rapid, reliable, and low-cost methods that enable discrimination among clinically relevant bacteria is crucial, with emphasis on those listed as WHO Global Priority 1 Critical Pathogens, such as carbapenem-resistant Acinetobacter baumannii and carbapenem-resistant or ESBL-producing Klebsiella pneumoniae. To address this problem, we developed and validated a protocol of surface-enhanced Raman spectroscopy (SERS) with silver nanostars for the discrimination of A. baumannii and K. pneumoniae species, and their globally disseminated and clinically relevant antibiotic resistant clones. Isolates were characterized by mixing bacterial colonies with silver nanostars, followed by deposition on filter paper for SERS spectrum acquisition. Spectral data were processed with unsupervised and supervised multivariate data analysis methods, including principal component analysis (PCA) and partial least-squares discriminant analysis (PLSDA), respectively. Our proposed SERS procedure using silver nanostars adsorbed to the bacteria, followed by multivariate data analysis, enabled differentiation between and within species. This pilot study demonstrates the potential of SERS for the rapid discrimination of clinically relevant A. baumannii and K. pneumoniae species and clones, displaying several advantages such as the ease of silver nanostars synthesis and the possible use of a handheld spectrometer, which makes this approach ideal for point-of-care applications.

Retzius-sparing robot-assisted radical prostatectomy in a medium size oncological center holds adequate oncological and functional outcomes.

Retzius-sparing robot-assisted radical prostatectomy (RS-RARP) has emerged as a surgical option for patients with prostatic cancer in high-volume centers. The objective is to assess oncological and functional outcomes when implementing RS-RARP in a medium-volume center without previous experience of robotic surgery. This is a prospective observational single-center study. Patients operated between July 2017 and April 2020 were divided into two consecutive groups, A and B, each with 104 patients. The surgeons had prior experience in laparoscopic surgery and underwent robotic training. Positive surgical margin (PSM) status, urinary continence, and erectile function projected by Kaplan-Meier curves, together with patient reported quality of life outcomes at 12 months post-surgery were documented. Median patient age was 63 years (IQR = 59-67), overall PSM rate were 33%, 28% for pT2 disease. Pre-operative values showed no significant difference between both groups. The rate of urinary continence dropped from 81 to 78% (SE = 5.7) (Group A) and from 90 to 72% (SE = 6.3) (Group B) using the International Consultation on Incontinence Questionnaire-Short Form. Baseline sexual function was regained in 41% (Group A) and 47% (Group B) of patients. The median Expanded Prostate Index Composite-26 total score decreased from 86 to 82. These outcomes relate favorably to prior reports. There was a clinically significant decrease in median operative time in the successive groups with post-operative complications occurring in less than 2% of surgical procedures overall. A 12-month follow-up suggests that RS-RARP may be safely introduced in a medium-volume center without previous experience of robotic surgery.

Nematode gene annotation by machine-learning-assisted proteotranscriptomics enables proteome-wide evolutionary analysis.

Nematodes encompass more than 24,000 described species, which were discovered in almost every ecological habitat, and make up >80% of metazoan taxonomic diversity in soils. The last common ancestor of nematodes is believed to date back to ∼650-750 million years, generating a large and phylogenetically diverse group to be explored. However, for most species high-quality gene annotations are incomprehensive or missing. Combining short-read RNA sequencing with mass spectrometry-based proteomics and machine-learning quality control in an approach called proteotranscriptomics, we improve gene annotations for nine genome-sequenced nematode species and provide new gene annotations for three additional species without genome assemblies. Emphasizing the sensitivity of our methodology, we provide evidence for two hitherto undescribed genes in the model organism Caenorhabditis elegans Extensive phylogenetic systems analysis using this comprehensive proteome annotation provides new insights into evolutionary processes of this metazoan group.

Inpatient rehabilitation of a person with Guillain-Barré syndrome associated with COVID-19 infection: An expert interdisciplinary approach to a case study.

BACKGROUND AND PURPOSE: Several cases of Guillain-Barré Syndrome (GBS) associated with a COVID-19 infection have been recently reported. Rehabilitation might be a key player in the recovery of these highly complex patients however, results are yet unknown. This case report aimed to describe the effects of an inpatient rehabilitation program, with an interdisciplinary team approach, in a patient with GBS in the context of a COVID-19 infection. CASE DESCRIPTION: A 58-year-old man with GBS after COVID-19 started an inpatient rehabilitation program focused on reducing dyspnea and fatigue symptoms; improving muscle strength, balance, aerobic and functional training; practicing activities of daily living and energy conservation techniques; swallowing training; emotional support and patient and family education about daily routines. An expert interdisciplinary team delivered the intervention, approximately 5 h/day, 5 days/week for 6 weeks. OUTCOMES: Improvements were observed in dyspnea, fatigue, nocturnal ventilation, muscle strength, balance, walking capacity, functional status, and swallowing function. CONCLUSION: This clinical case report illustrates the impact of a tailored and interdisciplinary rehabilitation program, on promoting recovery in multiple health domains of a patient with GBS associated with COVID-19 infection. Our experience might be useful to guide other inpatient rehabilitation programs to successfully manage these highly complex patients.

Inpatient rehabilitation can improve functional outcomes of post-intensive care unit COVID-19 patients-a prospective study.

PURPOSE: To evaluate the impact of an inpatient multimodal and intensive rehabilitation program on neuromuscular, respiratory, and functional impairments of post-ICU COVID-19 patients. MATERIALS AND METHODS: Prospective study including post-ICU COVID-19 survivors consecutively admitted to a rehabilitation centre. Rehabilitation was conducted by an interdisciplinary team. Medical Research Council (MRC) score, maximum expiratory pressure (MEP), maximum inspiratory pressure (MIP), peak cough flow (PCF), Functional Oral Intake Scale (FOIS), Brief Balance Evaluation Systems Test (Brief-BESTest), Timed Up and Go (TUG) test, 1 min Sit to Stand Test (1' STST), 6 min Walking Test (6MWT), Fatigue Assessment Scale (FAS), Functional Independence Measure (FIM) were assessed at admission (T0) and discharge (T1). RESULTS: A total of 42 patients were included. After 32.00;26.00 days of inpatient rehabilitation, there was a significant improvement in limb and respiratory muscle strength, cough effectiveness, fatigue, balance, exercise capacity, and in the ability to perform activities of daily living. Advanced age, longer acute care hospitalization, depressive symptoms, and cognitive deficits were associated with poorer functional outcomes. CONCLUSION: Post-ICU COVID-19 patients present multiple sequelae with detrimental functional impact. An adapted interdisciplinary rehabilitation program is essential for a thorough evaluation of these patients and results in significant functional gains.IMPLICATIONS FOR REHABILITATIONPost-ICU COVID-19 survivors present multiple sequelae and disabilities.An intensive and interdisciplinary inpatient rehabilitation results in significant improvement in limb and respiratory muscle strength, cough effectiveness, fatigue, balance, exercise capacity, and ability to perform activities of daily living.Timely referral from the acute care setting to rehabilitation services is crucial to minimize the functional impact of severe multisystemic disease and prolonged hospitalization.

Post-intensive Care Unit COVID-19 Survivors: Functional Status and Respiratory Function Three Months After an Inpatient Rehabilitation Program.

Introduction and objectives Long-term coronavirus disease 2019 (COVID-19) sequelae have become an increasing concern, with persistent dyspnoea and fatigue being the most common and long-lasting symptoms reported. The aim of this study was to evaluate the functional status and respiratory function three months after discharge from an inpatient rehabilitation program. Materials and methods This was a prospective study including post-ICU COVID-19 survivors consecutively admitted to an inpatient and multimodal rehabilitation program in a rehabilitation center. Evaluation of functional status (brief balance evaluation systems test (brief-BESTEST), timed up and go (TUG) test, 1 min sit to stand test (1STST), 6 min walking test (6MWT)); respiratory muscle strength (maximum expiratory pressure (MEP), maximum inspiratory pressure (MIP)); cough effectiveness (peak cough flow (PCF)); and fatigue (fatigue assessment scale (FAS)) were assessed at admission (T0), discharge (T1), and three months after discharge (T2). Results A total of 36 patients were included. Between T1 and T2, there was a significant improvement in MEP (84.47±20.89 vs 97.23±24.63 cmH2O, p<0.001), PCF (367.83±117.24 vs 441.33±132.90 L/min, p=0.003), functional capacity (1STST (19.90±6.37 vs 23.13±6.07, p=0.004), and 6MWT (459.25±153.70 vs 500.00±163.74 meters, p=0.003)). No differences were seen in MIP, brief-BESTEST, or TUG. Patients presented a higher median final FAS score at T2 compared to T1: 21.50±5 vs 18.60±2.65, p=0.002. Conclusions Post-ICU COVID-19 survivors admitted to an inpatient rehabilitation program maintained a good functional recovery at the three-month follow-up. Despite overall improvement, we found higher scores of FAS, suggesting worse fatigue levels.

Cystic lymphangioma of the pancreatic head treated by enucleation: Case report and literature review.

INTRODUCTION: The widespread use of imaging methods has led to an increased identification of asymptomatic Pancreatic Cystic Lymphangiomas (PCL), a rare entity for which available information is very limited. PRESENTATION OF CASE: We present the case of an asymptomatic 61-year-old male, submitted to elective enucleation of a pancreatic head PCL at our institution. After four years of follow-up the patient is doing well and has no clinical or imaging signs of recurrence. DISCUSSION: Though rare, PCL should be included in the differential diagnosis of pancreatic cystic neoplasms. All efforts should be made to ascertain a preoperative diagnosis, as expectant follow-up could be a reasonable approach in asymptomatic patients and/or poor surgical candidates. In the face of an uncertain diagnosis, complete surgical excision may be the treatment of choice. CONCLUSION: The medical community worldwide should be encouraged to report all cases of PCL, as to increment the overall knowledge about this lesion.

The Caenorhabditis elegans TDRD5/7-like protein, LOTR-1, interacts with the helicase ZNFX-1 to balance epigenetic signals in the germline.

LOTUS and Tudor domain containing proteins have critical roles in the germline. Proteins that contain these domains, such as Tejas/Tapas in Drosophila, help localize the Vasa helicase to the germ granules and facilitate piRNA-mediated transposon silencing. The homologous proteins in mammals, TDRD5 and TDRD7, are required during spermiogenesis. Until now, proteins containing both LOTUS and Tudor domains in Caenorhabditis elegans have remained elusive. Here we describe LOTR-1 (D1081.7), which derives its name from its LOTUS and Tudor domains. Interestingly, LOTR-1 docks next to P granules to colocalize with the broadly conserved Z-granule helicase, ZNFX-1. The Tudor domain of LOTR-1 is required for its Z-granule retention. Like znfx-1 mutants, lotr-1 mutants lose small RNAs from the 3' ends of WAGO and mutator targets, reminiscent of the loss of piRNAs from the 3' ends of piRNA precursor transcripts in mouse Tdrd5 mutants. Our work shows that LOTR-1 acts with ZNFX-1 to bring small RNA amplifying mechanisms towards the 3' ends of its RNA templates.

Taming transposable elements in vertebrates: from epigenetic silencing to domestication.

Transposable element (TE)-derived sequences are ubiquitous in most eukaryotic genomes known to date. Because their expression and mobility can lead to genomic instability, several pathways have evolved to control TEs. Nevertheless, TEs represent an important source of genomic novelty and are often co-opted for novel functions that are relevant for phenotypic divergence and adaptation. Here, we review how animals, in particular vertebrates, mitigate TE mobility and expression, alongside known examples of TE domestication. We argue that the next frontier is to understand the determinants and dynamics of TE domestication: how they shift from 'non-self' targets of epigenetic silencing to 'self' genetic elements. New technologies enable avenues of research that may close the gap between epigenetic silencing and domestication of TEs.

Nanoemulsion of cashew gum and clove essential oil (Ocimum gratissimum Linn) potentiating antioxidant and antimicrobial activity.

In this study, nanoemulsions of essential oil from Ocimumgratissimum (Linn) (EO) were produced using low and high energy techniques using cashew gum (CG) as a co-surfactant. The main constituents of the EO were determined by Gas Chromatography coupled with Mass Spectrometry (GC-MS), and their presence in the EO and in the formulations verified by Fourier Transform Infrared Spectroscopy (FTIR) and UV-visible spectrophotometry was observed the encapsulation efficiency (EE%), with colloidal stability. Nuclear magnetic resonance (NMR) was used to study cashew gum. Dynamic light scattering analysis (DLS) determined the nanoemulsion Z means, polydispersity index and the Zeta potential value, nanoparticle tracking analysis (NTA) were determined. The nanostructured EO showed better antibacterial action against the pathogenic gastroenteritis species Staphylococcus aureus, Escherichia coli and Salmonella enterica when compared to free EO. Atomic Force Microscopy (AFM) was used for morphological analysis of the nanoparticle and study of the action of the nanoemulsion through images of the cellular morphology of S. enterica. The antioxidant activity was evaluated against the ABTS radical (2,2'-azino-bis diazonium salt (3-ethylbenzothiazoline-6-sulfonic acid)). The encapsulation of EO in a nanostructured system improved its antibacterial and antioxidant activity, the low energy synthesis showed greater storage stability, remaining stable for 37 days.

Reusable and highly sensitive SERS immunoassay utilizing gold nanostars and a cellulose hydrogel-based platform.

The development of robust and sensitive point-of-care testing platforms is necessary to improve patient care and outcomes. Surface-enhanced Raman scattering (SERS)-based immunosensors are especially suited for this purpose. Here, we present a highly sensitive and selective SERS immunoassay, demonstrating for example the detection of horseradish peroxidase (HRP), in a sandwich format. The strength of our biosensor lies in merging: (i) SERS-immunotags based on gold nanostars, allowing exceptional intense SERS from attached Raman probes, covalent attachment of anti-HRP antibodies by a simple chemical method providing exceptional antigen binding activity; (ii) the ease of preparation of the capture platform from a regenerated cellulose-based hydrogel, a transparent material, ideal for microfluidics applications, with low background fluorescence and Raman signal, particularly suited for preserving high activity of the covalently bound anti-HRP antibodies. The sandwich complexes formed were characterised by atomic force microscopy, and by scanning electron microscopy coupled with electron diffraction spectroscopy; and (iii) the robustness of the simple Classical Least Squares method for SERS data analysis, resulting in superior discrimination of SERS signals from the background and much better data fitting, compared to the commonly used peak integral method. Our SERS immunoassay greatly improves the detection limits of traditional enzyme-linked immunosorbent assay approaches, and its performance is better or comparable to those of existing SERS-based immunosensors. Our approach successfully overcomes the main challenges of application at point-of-care, including increasing reproducibility, sensitivity, and specificity, associated with an environmentally friendly and robust design. Also, the proposed design withstands several cycles of regeneration, a feature absent in paper-SERS immunoassays and this opens the way for sensitive multiplexing applications on a microfluidic platform.

Acetylated cashew gum and fucan for incorporation of lycopene rich extract from red guava (Psidium guajava L.) in nanostructured systems: Antioxidant and antitumor capacity.

Industrial application of lycopene is limited due to its chemical instability and low bioavailability. This study proposes the development of fucan-coated acetylated cashew gum nanoparticles (NFGa) and acetylated cashew gum nanoparticles (NGa) for incorporation of the lycopene-rich extract from red guava (LEG). Size, polydispersity, zeta potential, nanoparticles concentration, encapsulation efficiency, transmission electron microscopy (TEM) and atomic force microscopy (AFM) were used to characterize nanoparticles. The antioxidant activity was determinated and cell viability was evaluated in the human breast cancer cells (MCF-7) and human keratinocytes (HaCaT) by MTT assay. The toxic effect was evaluated by hemolysis test and by Galleria mellonella model. NFGa showed higher stability than NGa, having a size of 162.10 ± 3.21 nm, polydispersity of 0.348 ± 0.019, zeta potential -30.70 ± 0.53 mV, concentration of 6.4 × 109 nanoparticles/mL and 60% LEG encapsulation. Microscopic analysis revealed a spherical and smooth shape of NFGa. NFGa showed antioxidant capacity by ABTS method and ORAC assay. The NFGa presented significant cytotoxicity against MCF-7 from the lowest concentration tested (6.25-200 µg/mL) and did not affect the cell viability of the HaCaT. NFGa showed non-toxic effect in the in vitro and in vivo models. Therefore, NFGa may have a promising application in LEG stabilization for antioxidant and antitumor purposes.