Direct Zeptomole Detection of RNA Biomarkers by Ultrabright Fluorescent Nanoparticles on Magnetic Beads.

Nucleic acids are important biomarkers in cancer and viral diseases. However, their ultralow concentration in biological/clinical samples makes direct target detection challenging, because it leads to slow hybridization kinetics with the probe and its insufficient signal-to-noise ratio. Therefore, RNA target detection is done by molecular (target) amplification, notably by RT-PCR, which is a tedious multistep method that includes nucleic acid extraction and reverse transcription. Here, a direct method based on ultrabright dye-loaded polymeric nanoparticles in a sandwich-like hybridization assay with magnetic beads is reported. The ultrabright DNA-functionalized nanoparticle, equivalent to ≈10 000 strongly emissive rhodamine dyes, is hybridized with the magnetic bead to the RNA target, providing the signal amplification for the detection. This concept (magneto-fluorescent sandwich) enables high-throughput detection of DNA and RNA sequences of varied lengths from 48 to 1362 nt with the limit of detection down to 0.3 fm using a plate reader (15 zeptomoles), among the best reported for optical sandwich assays. Moreover, it allows semi-quantitative detection of SARS-CoV-2 viral RNA directly in clinical samples without a dedicated RNA extraction step. The developed technology, combining ultrabright nanoparticles with magnetic beads, addresses fundamental challenges in RNA detection; it is expected to accelerate molecular diagnostics of diseases.

Characterization of expression patterns and dynamic relocation of Notch proteins during acrosome reaction of bull spermatozoa.

Notch is a conserved cell-signaling pathway involved in spermatogenesis regulation. This study firstly evaluated the presence, localization patterns, acquisition origin and relation to acrosome reaction of Notch proteins in bull sperm. Western Blot analysis detected all Notch proteins in ejaculated bull sperm, and immunostaining described their specific sperm localization. Recovery of sperm from different segments showed that Notch proteins have testicular origin (NOTCH1, NOTCH2, DLL4), are sequentially acquired during sperm maturation along epididymal transit (NOTCH3, DLL3, JAGGED1-2), or post-ejaculation (DLL1, NOTCH4). Testis NOTCH2 is ubiquitously expressed in all germ-cell lines, whereas DLL4 is expressed in round and elongated spermatids during the Golgi, Cap, Acrosome and Maturation phases. In vitro spontaneous and induced sperm acrosome reaction induce consistent sperm regional relocation of NOTCH2, DLL4 and JAGGED1, and these relocation patterns are significantly associated to sperm acrosome status. NOTCH2 and JAGGED1 are relocated from the head apical to the post-equatorial regions, whereas DLL4 is lost along with the acrosome, evidencing that sperm spatial redistribution of NOTCH2 and JAGGED1 is linked to acrosome reaction onset, whereas DLL4 loss is linked to AR completion. Overall, results prompt for a relevant Notch role in bull sperm acrosome testicular development, epididymal maturation and acrosome reaction.

First Insights on Resistance and Virulence Potential of Escherichia coli from Captive Birds of Prey in Portugal.

Captive birds of prey are often used for pest control in urban areas, while also participating in falconry exhibitions. Traveling across the country, these birds may represent a public health concern as they can host pathogenic and zoonotic agents and share the same environment as humans and synanthropic species. In this work, Escherichia coli from the cloacal samples of 27 captive birds of prey were characterized to determine their pathogenic potential. Isolates were clustered through ERIC-PCR fingerprinting, and the phylogenetic groups were assessed using a quadruplex PCR method. Their virulence and resistance profile against nine antibiotics were determined, as well as the isolates' ability to produce extended-spectrum ß-lactamases (ESBLs). The 84 original isolates were grouped into 33 clonal types, and it was observed that more than half of the studied isolates belonged to groups D and B2. Most isolates presented gelatinase activity (88%), almost half were able to produce biofilm (45%), and some were able to produce α-hemolysin (18%). The isolates presented high resistance rates towards piperacillin (42%), tetracycline (33%), and doxycycline (30%), and 6% of the isolates were able to produce ESBLs. The results confirm the importance of these birds as reservoirs of virulence and resistance determinants that can be disseminated between wildlife and humans, stressing the need for more studies focusing on these animals.

Design of a multi-epitope-based vaccine candidate against Bovine Genital Campylobacteriosis using a reverse vaccinology approach.

BACKGROUND: Bovine Genital Campylobacteriosis (BGC), a worldwide distributed venereal disease caused by Campylobacter fetus subsp. venerealis (Cfv), has a relevant negative economic impact in cattle herds. The control of BGC is hampered by the inexistence of globally available effective vaccines. The present in silico study aimed to develop a multi-epitope vaccine candidate against Cfv through reverse vaccinology. RESULTS: The analysis of Cfv strain NCTC 10354 proteome allowed the identification of 9 proteins suitable for vaccine development. From these, an outer membrane protein, OmpA, and a flagellar protein, FliK, were selected for prediction of B-cell and T-cell epitopes. The top-ranked epitopes conservancy was assessed in 31 Cfv strains. The selected epitopes were integrated to form a multi-epitope fragment of 241 amino acids, which included 2 epitopes from OmpA and 13 epitopes from FliK linked by GPGPG linkers and connected to the cholera toxin subunit B by an EAAAK linker. The vaccine candidate was predicted to be antigenic, non-toxic, non-allergenic, and soluble upon overexpression. The protein structure was predicted and optimized, and the sequence was successfully cloned in silico into a plasmid vector. Additionally, immunological simulations demonstrated the vaccine candidate's ability to stimulate an immune response. CONCLUSIONS: This study developed a novel vaccine candidate suitable for further in vitro and in vivo experimental validation, which may become a useful tool for the control of BGC.

Determinants of HIV late presentation among men who have sex with men in Portugal (2014-2019): who's being left behind?

Introduction: HIV late presentation (LP) remains excessive in Europe. We aimed to analyze the factors associated with late presentation in the MSM population newly diagnosed with HIV in Portugal between 2014 and 2019. Methods: We included 391 newly HIV-1 diagnosed Men who have Sex with Men (MSM), from the BESTHOPE project, in 17 countrywide Portuguese hospitals. The data included clinical and socio-behavioral questionnaires and the viral genomic sequence obtained in the drug resistance test before starting antiretrovirals (ARVs). HIV-1 subtypes and epidemiological surveillance mutations were determined using different bioinformatics tools. Logistic regression was used to estimate the association between predictor variables and late presentation (LP). Results: The median age was 31 years, 51% had a current income between 501-1,000 euros, 28% were migrants. 21% had never been tested for HIV before diagnosis, with 42.3% of MSM presenting LP. 60% were infected with subtype B strains. In the multivariate regression, increased age at diagnosis, higher income, lower frequency of screening, STI ever diagnosed and higher viral load were associated with LP. Conclusion: Our study suggests that specific subgroups of the MSM population, such older MSM, with higher income and lower HIV testing frequency, are not being targeted by community and clinical screening services. Overall, targeted public health measures should be strengthened toward these subgroups, through strengthened primary care testing, expanded access to PrEP, information and promotion of HIV self-testing and more inclusive and accessible health services.

The effect of common mental disorders on suicidal attempts by women, during pregnancy and six to nine years after birth. / Efeito dos transtornos mentais comuns na gestação e seis a nove anos pós-parto para a tentativa de suicídio em mulheres.

There is scarcity of global data on suicide attempts (SA). The higher frequency of mental disorders places women at a higher risk for SA. Mental disorders (MD) are ranked first in the Global Burden of Diseases (GBD) in terms of years lived with disability (YLD) and common mental disorders (CMD) affect women the most. IN this light, the present study aimed to investigate the impact of CMD during pregnancy, as well as 6 to 9 years after birth, on SA in women who were registered in the Family Health Strategy from the town of Recife, Pernambuco, Brazil. This study consisted of two stages of a prospective cohort. 643 adult women were included. The prevalence of CMD in stage I (pregnancy) and in stage III (six to nine years after birth) were, respectively, 19.3% and 12.6%. The incidence of SA was 10.9%. The multivariate analysis demonstrated a cumulative effect of CMD for SA: only during pregnancy (OR 5.4; 95%CI 2.2-13.3); only in the third stage (OR 5.8; 95%CI 2.3-14.9); and in both stages (OR 6.0; 95%CI 2.5-14.4). The increase of CMD in women increases the chances of SA, hence the importance of implementing public policies for women's health, especially for those with a history of mental disorders and unhealthy habits, and those who suffer violence.

Há escassez de dados globais sobre as tentativas de suicídio (TS). A maior frequência de adoecimento mental coloca as mulheres em maior risco de TS. Os transtornos mentais (TM) estão em primeiro lugar na carga global de doenças em termos de anos vividos com incapacidade (AVI). Entre os problemas de saúde mental que mais acometem mulheres estão os transtornos mentais comuns (TMC). O objetivo desta pesquisa foi investigar o impacto dos TMC na gravidez e seis a nove anos após o parto para a TS em mulheres cadastradas na Estratégia de Saúde da Família no Recife, Pernambuco, Brasil. O estudo engloba duas etapas de uma coorte prospectiva. Foram incluídas 643 mulheres adultas. A prevalência de TMC na etapa I (gestação) e sua incidência na etapa III (seis a nove anos após o parto) foram, respectivamente, 19,3% e 12,6%. A incidência da TS foi de 10,9%. A análise multivariada demonstrou efeito cumulativo dos TMC para a TS: TMC só na gestação (OR 5,4; IC95% 2,2-13,3); só na terceira etapa (OR 5,8; IC95% 2,3-14,9); e em ambas (OR 6,0; IC95% 2,5-14,4). O acúmulo dos TMC em mulheres aumenta a chance de TS, sendo importante a implementação de políticas públicas para a saúde das mulheres, principalmente com histórico de doença mental, hábitos não saudáveis e que sofrem violência.

Efeito dos transtornos mentais comuns na gestação e seis a nove anos pós-parto para a tentativa de suicídio em mulheres / The effect of common mental disorders on suicidal attempts by women, during pregnancy and six to nine years after birth

Resumo Há escassez de dados globais sobre as tentativas de suicídio (TS). A maior frequência de adoecimento mental coloca as mulheres em maior risco de TS. Os transtornos mentais (TM) estão em primeiro lugar na carga global de doenças em termos de anos vividos com incapacidade (AVI). Entre os problemas de saúde mental que mais acometem mulheres estão os transtornos mentais comuns (TMC). O objetivo desta pesquisa foi investigar o impacto dos TMC na gravidez e seis a nove anos após o parto para a TS em mulheres cadastradas na Estratégia de Saúde da Família no Recife, Pernambuco, Brasil. O estudo engloba duas etapas de uma coorte prospectiva. Foram incluídas 643 mulheres adultas. A prevalência de TMC na etapa I (gestação) e sua incidência na etapa III (seis a nove anos após o parto) foram, respectivamente, 19,3% e 12,6%. A incidência da TS foi de 10,9%. A análise multivariada demonstrou efeito cumulativo dos TMC para a TS: TMC só na gestação (OR 5,4; IC95% 2,2-13,3); só na terceira etapa (OR 5,8; IC95% 2,3-14,9); e em ambas (OR 6,0; IC95% 2,5-14,4). O acúmulo dos TMC em mulheres aumenta a chance de TS, sendo importante a implementação de políticas públicas para a saúde das mulheres, principalmente com histórico de doença mental, hábitos não saudáveis e que sofrem violência.

Abstract There is scarcity of global data on suicide attempts (SA). The higher frequency of mental disorders places women at a higher risk for SA. Mental disorders (MD) are ranked first in the Global Burden of Diseases (GBD) in terms of years lived with disability (YLD) and common mental disorders (CMD) affect women the most. IN this light, the present study aimed to investigate the impact of CMD during pregnancy, as well as 6 to 9 years after birth, on SA in women who were registered in the Family Health Strategy from the town of Recife, Pernambuco, Brazil. This study consisted of two stages of a prospective cohort. 643 adult women were included. The prevalence of CMD in stage I (pregnancy) and in stage III (six to nine years after birth) were, respectively, 19.3% and 12.6%. The incidence of SA was 10.9%. The multivariate analysis demonstrated a cumulative effect of CMD for SA: only during pregnancy (OR 5.4; 95%CI 2.2-13.3); only in the third stage (OR 5.8; 95%CI 2.3-14.9); and in both stages (OR 6.0; 95%CI 2.5-14.4). The increase of CMD in women increases the chances of SA, hence the importance of implementing public policies for women's health, especially for those with a history of mental disorders and unhealthy habits, and those who suffer violence.

Cerebral arterial gas embolism (CAGE) during open water scuba certification training whilst practising a controlled emergency swimming ascent.

We report the case of a 23-year-old male novice diver who sustained cerebral arterial gas embolism (CAGE) during his open water certification training whilst practising a free ascent as part of the course. He developed immediate but transient neurological symptoms that had resolved on arrival to hospital. Radiological imaging of his chest showed small bilateral pneumothoraces, pneumopericardium and pneumomediastinum. In view of this he was treated with high flow normobaric oxygen rather than recompression, because of the risk of development of tension pneumothorax upon chamber decompression. There was no relapse of his neurological symptoms with this regimen. The utility and safety of free ascent training for recreational divers is discussed, as is whether a pneumothorax should be vented prior to recompression, as well as return to diving following pulmonary barotrauma.

Impact of Structural GLA Protein Changes on Peripheral GLA Activity and Substrate Accumulation in Fabry Disease Patients.

INTRODUCTION: Fabry disease is an X-linked lysosomal storage disorder caused by pathogenic variants in the GLA gene, leading to decreased/absent α-galactosidase activity. In clinical practice, enzyme activity and substrate/byproduct accumulation play a role in diagnosis and disease-monitoring biomarkers. However, interpreting biomarker levels is not straightforward and can change according to the underlying GLA protein abnormality. OBJECTIVE: Our goals were to understand how disrupting specific protein regions changes biomarker behaviour and to establish specific patterns for individual variants. METHODOLOGY: We analysed data from the Biochemical Genetics Laboratory regarding GLA variants, GLA enzyme activity (in dried blood spots, plasma or white blood cells), plasma LysoGb3 accumulation, and urinary Gb3 excretion. We assessed correlations, trends, and potential predictor models of biomarker behaviour. RESULTS: We assessed 169 hemizygous male and 255 heterozygous female patients. For both groups, substrate accumulation correlates inversely with GLA activity. Variants affecting residues buried within the protein core or the active site were associated with more severe biomarker changes, while those affecting residues that establish disulfide bonds or are glycosylated were similar to other variants. For each non-truncating variant, we also established specific profiles of biomarker behaviour. Finally, we also designed predictor models of biomarker behaviour based on structural variant information. This study provides the groundwork for the impact of GLA protein variation on GLA activity and substrate accumulation. CONCLUSION: This knowledge is of extreme relevance for diagnostic labs and clinicians, as some genetic variants are challenging to interpret regarding pathogenicity. Assessing whether biomarker changes are in the expected range for a specific variant may help diagnostic evaluation. This study also contributes to recognising non-disease-causing variants, considering their overall biochemical impact, and providing a comparative reference for biomarker discovery studies. In the future, the correlation of these findings with disease severity may be of great relevance for diagnosis and monitoring progression.

Insights into the antimicrobial properties of a cationic steroid and antibiofilm performance in PDMS-based coatings to potentially treat urinary infections.

Currently, multidrug-resistant (MDR) infections are one of the most important threats, driving the search for new antimicrobial compounds. Cationic peptide antibiotics (CPAs) and ceragenins (CSAs) contain in their structures cationic groups and adopt a facially amphiphilic conformation, conferring the ability to permeate the membranes of bacteria and fungi. Keeping these features in mind, an amine steroid, DOCA-NH2, was found to be active against reference strains and MDR isolates of Gram-positive Enterococcus faecalis and Staphylococcus aureus and Gram-negative Escherichia coli and Pseudomonas aeruginosa. The compound was active against all the tested microorganisms, having bactericidal and fungicidal activity, displaying minimal inhibitory concentrations (MICs) between 16 and 128 µg mL-1. No synergy with clinically relevant antibacterial drugs was found. However, the compound was able to completely inhibit the biofilm formation of bacteria exposed to the MIC of the compound. For E. coli and E. faecalis, inhibition of biofilm formation occurred at half the MIC. Besides, DOCA-NH2 inhibited the dimorphic transition of Candida albicans at concentrations 4 times lower than the MIC, and can reduce the microorganism virulence and biofilm formation was significantly reduced at both MIC and half the MIC. Polydimethylsiloxane-based coatings containing DOCA-NH2 (0.5, 1.0, and 1.5 wt%) were prepared and tested against the E. coli biofilm formation under hydrodynamic conditions similar to those prevailing in ureteral stents. A biofilm reduction of approximately 80% was achieved when compared to the control.

Characterization of circulating microRNA profiles of postpartum dairy cows with persistent subclinical endometritis.

Subclinical endometritis (SCE) is an unresolved inflammation of the endometrium of postpartum dairy cows, seriously affecting fertility. Current diagnosis, which relies on uterine cytology or even more invasive biopsy sampling, would benefit from the identification of blood-based diagnostic biomarkers. Due to the known role of microRNAs (miRNAs) in other diseases, this case-control study evaluated the cell-free circulating miRNA profiles of SCE cows, and the network of transcripts predicted to interact with those miRNAs, previously identified as differentially expressed genes (DEG) in the endometrium of the same cows. Healthy (H, n = 6) and persistent SCE (n = 11) cows characterized by endometrial cytology and biopsy were blood sampled at 21 and 44 d postpartum (DPP). Following extraction of cell-free plasma miRNAs and RNA-seq analysis, differential abundance analysis of miRNAs was performed with the DESeq2 R package (adjusted p-value of 0.05), and in silico prediction of miRNA-interacting genes on a sequence complementary basis was conducted using the miRWalk database. The principal component analysis showed a clear clustering between groups of uterine health phenotypes (H vs. SCE), although the clustering between groups was less pronounced at 44 DPP than at 21 DPP. No effect of the stage (21 vs. 44 DPP) was observed. A total of 799 known circulating miRNAs were identified, from which 34 demonstrated differential abundance between H and SCE cows (12 less abundant and 22 more abundant in SCE than in H cows). These 34 miRNAs are predicted to interact with 10,104 transcripts, among which 43, 81, and 147 were previously identified as differentially expressed in, respectively, endometrial luminal epithelial, glandular epithelial, and stromal cells of the same cows. This accounts for approximately half of the DEG identified between those H and SCE cows, including genes involved in endometrial cell proliferation, angiogenesis and immune response, whose dysregulation in SCE cows may impair pregnancy establishment. From 219 miRNAs with mean normalized read counts above 100, the presence and abundance of miR-425-3p and miR-2285z had the highest discriminatory level to differentiate SCE from H cows. In conclusion, despite apparent confinement to the endometrium, SCE is associated with a distinct circulating miRNA profile, which may represent a link between the systemic changes associated with disease and the endometrial immune response. The validation of a miRNA panel consisting of circulating cell-free miR-425-3p and miR-2285z may prove a relevant advancement for the noninvasive diagnosis of persistent SCE.

Could children exposed to intimate partner violence against their mother have more functional gastrointestinal disorders?

OBJECTIVE: To analyze the association between child exposure to intimate partner violence (CEIPV) committed against their mother and functional gastrointestinal disorders (FGIDs) in school-age children. METHODS: A prospective cohort study was carried out that evaluated 626 mother-child pairs living in a poor urban area in the Northeast of Brazil. The mothers were asked about their children's exposure to intimate partner violence. Holden's classification was used, which investigates ten types of CEIPV from prenatal care. The number of exposures, age of onset, and the perpetrator of the exposure were also verified. The FGIDs in children were identified from the mother's responses to the questionnaire about pediatric gastrointestinal symptoms, Roma III version parent-report, translated and cross-culturally adapted into Brazilian Portuguese. RESULTS: FGIDs were more frequent among children exposed to violence than those not exposed (OR: 1.64; 95% CI: 1.04-2.58; p = 0.03). Likewise, in children who suffered four or more types of exposure (OR: 1.81; 95% CI: 1.10-2.96; p  = 0.01), in which the exposure started in the first two years of life (OR: 1.93; 95% CI: 1.08-3.47; p = 0.02) and in those whose biological father was the perpetrator of the violence (OR; 1.62; 95% CI: 1, 05-2.48; p = 0.02). CONCLUSION: Corroborating the proposal of the biopsychosocial model, CEIPV increases the chance of occurrence of FGIDs in school-age children. Therefore, early identification and proposals for appropriate interventions are suggested both for the prevention of CEIPV and for the prevention and follow-up of FGIDs.

3,4-Dioxygenated xanthones as antifouling additives for marine coatings: in silico studies, seawater solubility, degradability, leaching, and antifouling performance.

Marine biofouling pollution is a process that impacts ecosystems and the global economy. On the other hand, traditional antifouling (AF) marine coatings release persistent and toxic biocides that accumulate in sediments and aquatic organisms. To understand the putative impact on marine ecosystems of recently described and patented AF xanthones (xanthones 1 and 2), able to inhibit mussel settlement without acting as biocides, several in silico environmental fate predictions (bioaccumulation, biodegradation, and soil absorption) were calculated in this work. Subsequently, a degradation assay using treated seawater at different temperatures and light exposures was conducted for a period of 2 months to calculate their half-life (DT50). Xanthone 2 was found to be non-persistent (DT50 < 60 days) at 50 µM, contrary to xanthone 1 (DT50 > 60 days). To evaluate the efficacy of both xanthones as AF agents, they were blended into four polymeric-based coating systems: polyurethane- and polydimethylsiloxane (PDMS)-based marine paints, as well as room-temperature-vulcanizing PDMS- and acrylic-based coatings. Despite their low water solubility, xanthones 1 and 2 demonstrated suitable leaching behaviors after 45 days. Overall, the generated xanthone-based coatings were able to decrease the attachment of the Mytilus galloprovincialis larvae after 40 h. This proof-of-concept and environmental impact evaluation will contribute to the search for truly environmental-friendly AF alternatives.

Nisin Z Potential for the Control of Diabetic Foot Infections Promoted by Pseudomonas aeruginosa Persisters.

Diabetic foot ulcers (DFU) are a major complication of diabetes mellitus and a public health concern worldwide. The ability of P. aeruginosa to form biofilms is a key factor responsible for the chronicity of diabetic foot infections (DFIs) and frequently associated with the presence of persister cells. These are a subpopulation of phenotypic variants highly tolerant to antibiotics for which new therapeutic alternatives are urgently needed, such as those based on antimicrobial peptides. This study aimed to evaluate the inhibitory effect of nisin Z on P. aeruginosa DFI persisters. To induce the development of a persister state in both planktonic suspensions and biofilms, P. aeruginosa DFI isolates were exposed to carbonyl cyanide m-chlorophenylhydrazone (CCCP) and ciprofloxacin, respectively. After RNA extraction from CCCP-induced persisters, transcriptome analysis was performed to evaluate the differential gene expression between the control, persisters, and persister cells exposed to nisin Z. Nisin Z presented a high inhibitory effect against P. aeruginosa persister cells but was unable to eradicate them when present in established biofilms. Transcriptome analysis revealed that persistence was associated with downregulation of genes related to metabolic processes, cell wall synthesis, and dysregulation of stress response and biofilm formation. After nisin Z treatment, some of the transcriptomic changes induced by persistence were reversed. In conclusion, nisin Z could be considered as a potential complementary therapy for treating P. aeruginosa DFI, but it should be applied as an early treatment or after wound debridement.

Long-Range Energy Transfer between Dye-Loaded Nanoparticles: Observation and Amplified Detection of Nucleic Acids.

Förster resonance energy transfer (FRET) is essential in optical materials for light-harvesting, photovoltaics, and biosensing, but its operating range is fundamentally limited by the Förster radius of ≈5 nm. In this work, FRET between fluorescent organic nanoparticles (NPs) is studied in order to break this limit. The donor and acceptor NPs are built from charged hydrophobic polymers loaded with cationic dyes and bulky hydrophobic counterions. Their surface is functionalized with DNA in order to control surface-to-surface distance. It is found that the FRET efficiency does not follow the canonic Förster law, reaching 0.70 and 0.45 values for NP-NP distances of 15 and 20 nm, respectively. This corresponds to the FRET efficiency decay as power four of the surface-to-surface NP-NP distance. Based on this long-distance FRET, a DNA nanoprobe is developed, where a target DNA fragment, encoding the cancer marker survivin, bringing together donor and acceptor NPs at ≈15 nm distance. In this nanoprobe, a single-molecular recognition results in unprecedented color switch for >5000 dyes, yielding a simple and fast assay with 18 attomoles limit of detection. Breaking the Förster distance limit for ultrabright NPs opens the route to advanced optical nanomaterials for amplified FRET-based biosensing.

Maternal educational practices and mental health disorders of school-age children

Abstract Objective: To investigate the association between emotional and behavioral difficulties in schoolchildren and maternal educational practices. Methods: Participants and Setting: 631 mother-child pairs were evaluated in a poor urban district in Recife, northeastern Brazil. Cross-sectional study carried out between 2013 and 2014. It integrates a prospective cohort study designed to investigate the consequences of intimate partner violence that occurred during pregnancy, postpartum, and seven years after birth for the physical and mental health of women and their children. Maternal educational practices were assessed using the Parent-Child Conflict Tactics Scale (CTSPC - mother-child version) and the child's behavioral and emotional disorders through the Strengths and Difficulties Questionnaire (SDQ) mother-version. The association was estimated through crude and adjusted prevalence ratios, using Poisson regression considering the 95% confidence interval. Results: The prevalence of violent maternal educational practices was 91.8% (572/631) being 89.7% (566/631) of psychological aggression and 75.6% of physical aggression, subdivided into corporal punishment (73.5%), physical maltreatment (35.8%) and severe physical maltreatment (1.7%). Corporal punishment (Adjusted PR 1.5; CI 95%: 1.1-2.1; p = 0.010) and severe physical maltreatment (Adjusted PR 1.9; CI 95%: 1.3-2.8; p = 0.002) were associated with emotional and behavioral difficulties in schoolchildren. Conclusion: The high prevalence of violent maternal educational practices, especially corporal punishment and severe physical maltreatment is associated with emotional and behavioral disorders in children. Therefore, it is necessary to interventions that promote parental support and effective use of non-violent discipline in conducting the educational process to establish healthier family relationships and to prevent/mitigate the impact of emotional and behavioral problems in children. © 2022 Sociedade Brasileira de Pediatria. Published by Elsevier Editora Ltda. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/licenses/by-nc-nd/4.0/).

Biotinylated Fluorescent Polymeric Nanoparticles for Enhanced Immunostaining.

The performance of fluorescence immunostaining is physically limited by the brightness of organic dyes, whereas fluorescence labeling with multiple dyes per antibody can lead to dye self-quenching. The present work reports a methodology of antibody labeling by biotinylated zwitterionic dye-loaded polymeric nanoparticles (NPs). A rationally designed hydrophobic polymer, poly(ethyl methacrylate) bearing charged, zwitterionic and biotin groups (PEMA-ZI-biotin), enables preparation of small (14 nm) and bright fluorescent biotinylated NPs loaded with large quantities of cationic rhodamine dye with bulky hydrophobic counterion (fluorinated tetraphenylborate). The biotin exposure at the particle surface is confirmed by Förster resonance energy transfer with dye-streptavidin conjugate. Single-particle microscopy validates specific binding to biotinylated surfaces, with particle brightness 21-fold higher than quantum dot-585 (QD-585) at 550 nm excitation. The nanoimmunostaining method, which couples biotinylated antibody (cetuximab) with bright biotinylated zwitterionic NPs through streptavidin, significantly improves fluorescence imaging of target epidermal growth factor receptors (EGFR) on the cell surface compared to a dye-based labeling. Importantly, cetuximab labeled with PEMA-ZI-biotin NPs can differentiate cells with distinct expression levels of EGFR cancer marker. The developed nanoprobes can greatly amplify the signal from labeled antibodies, and thus become a useful tool in the high-sensitivity detection of disease biomarkers.

The Contribution of the Sheep and the Goat Model to the Study of Ovarian Ageing.

Ovarian ageing stands as the major contributor towards fertility loss. As such, there is an urge for studies addressing the mechanisms that promote ovarian ageing and new strategies aiming to delay it. Recently, the presence of a unique population of multinucleated giant cells has been identified in the ovaries of reproductively aged mice. These cells have been considered hallmarks of ovarian ageing. However, up to date multinucleated giant cells have only been described in the ovaries of the mice. Therefore, the aim of the present work was to evaluate and characterize the presence of such hallmarks of ovarian ageing in the sheep and the goat. In this study, ovaries from juvenile (6 months) and mature animals (18-24 months) were used. The hematoxylin and eosin technique was performed to describe the ovarian morphology and evaluate the ovarian follicle reserve pool. Sudan black B staining and the detection of autofluorescence emission were used to identify and characterize the presence of multinucleated giant cells. Statistical analyses were performed with GraphPad Prism 9.0.0. A decrease in the follicle reserve pool and the presence of multinucleated giant cells, with lipofuscin accumulation and the emission of autofluorescence, were observed in the ovaries of the mature animals of both species. Our results support the interest in the use of the ovine and the caprine model, that share physiological and pathophysiological characteristics with humans, in future studies addressing ovarian ageing.

Maternal educational practices and mental health disorders of school-age children.

OBJECTIVE: To investigate the association between emotional and behavioral difficulties in schoolchildren and maternal educational practices. METHODS: Participants and Setting: 631 mother-child pairs were evaluated in a poor urban district in Recife, northeastern Brazil. Cross-sectional study carried out between 2013 and 2014. It integrates a prospective cohort study designed to investigate the consequences of intimate partner violence that occurred during pregnancy, postpartum, and seven years after birth for the physical and mental health of women and their children. Maternal educational practices were assessed using the Parent-Child Conflict Tactics Scale (CTSPC - mother-child version) and the child's behavioral and emotional disorders through the Strengths and Difficulties Questionnaire (SDQ) mother-version. The association was estimated through crude and adjusted prevalence ratios, using Poisson regression considering the 95% confidence interval. RESULTS: The prevalence of violent maternal educational practices was 91.8% (572/631) being 89.7% (566/631) of psychological aggression and 75.6% of physical aggression, subdivided into corporal punishment (73.5%), physical maltreatment (35.8%) and severe physical maltreatment (1.7%). Corporal punishment (Adjusted PR 1.5; CI 95%: 1.1-2.1; p = 0.010) and severe physical maltreatment (Adjusted PR 1.9; CI 95%: 1.3-2.8; p = 0.002) were associated with emotional and behavioral difficulties in schoolchildren. CONCLUSION: The high prevalence of violent maternal educational practices, especially corporal punishment and severe physical maltreatment is associated with emotional and behavioral disorders in children. Therefore, it is necessary to interventions that promote parental support and effective use of non-violent discipline in conducting the educational process to establish healthier family relationships and to prevent/mitigate the impact of emotional and behavioral problems in children.

The road to recovery: Sensing public opinion towards reopening measures with social media data in post-lockdown cities.

The COVID-19 pandemic has resulted in cities implementing lockdown measures, causing unprecedented disruption (e.g. school/shop/office closures) to urban life often extending over months. With the spread of COVID-19 now being relatively contained, many cities have started to ease their lockdown restrictions by phases. Following the phased recovery strategy proposed by the UK government following the first national lockdown, this paper utilises Greater London as its case study, selecting three main reopening measures (i.e., schools, shops and hospitality reopening). This paper applies sentiment analysis and topic modelling to explore public opinions expressed via Twitter. Our findings reveal that public attention towards the reopening measures reached a peak before the date of policy implementation. The attitudes expressed in discussing reopening measures changed from negative to positive. Regarding the discussed topics related to reopening measures, we find that citizens are more sensitive to early-stage reopening than later ones. This study provides a time-sensitive approach for local authorities and city managers to rapidly sense public opinion using real-time social media data. Governments and policymakers can make use of the framework of sensing public opinion presented herein and utilise it in leading their post-lockdown cities into an adaptive, inclusive and smart recovery.