Predicting attachment in Portuguese infants born very or extremely preterm: Understanding the roles of infant regulatory behavior, maternal sensitivity, and risk factors.

A growing body of research shows that early attachment relationships are foundational for children's later developmental and psychosocial outcomes. However, findings are mixed regarding whether preterm birth predicts later attachment, but insecurity is generally more prevalent among infants at higher medical and/or social/familial risk. This longitudinal study aimed to identify specific relational, familial/demographic, and perinatal predictors of attachment in a sample of 63 Portuguese infants born very or extremely preterm (VEPT, <32 gestational weeks) and their mothers from diverse socioeconomic backgrounds. One-third of the mothers had social/family risk factors (e.g., single parent, immigrant, unemployed, low education, and/or low income). At 3 months (corrected age), dyads were observed during social interaction in the Face-to-Face Still-Face paradigm (FFSF) and during free play. At 12 months, mother-infant dyads were observed in Ainsworth's Strange Situation. Over half (58.7%) of the infants were classified as insecurely attached. Social-Positive Oriented regulatory behavior pattern, higher maternal sensitivity, higher infant cooperation during free play, number of siblings and an absence of social/family risk factors were associated with attachment security. Perinatal variables were unrelated to attachment. Findings indicate that both relational and social contextual factors contribute to attachment in this biologically vulnerable sample.

Un creciente cuerpo investigativo muestra que las relaciones afectivas tempranas son fundamentales para posteriores resultados de desarrollo y sicosociales de los niños. Sin embargo, los resultados son variados acerca de si el nacimiento prematuro predice la afectividad posterior, pero la inseguridad es generalmente más prevalente entre infantes bajo más alto riesgo médico y/o social/familiar. Este estudio longitudinal se propuso identificar factores específicos de predicción de la afectividad, relacionales, familiar/demográficos y perinatales en un grupo muestra de 63 infantes portugueses nacidos muy o extremadamente prematuros (VEPT, < 32 semanas gestacionales) y sus madres de diversos niveles socioeconómicos. Un tercio de las madres tenían factores de riesgo social/familiar (v.g. madre soltera, inmigrante, desempleada, de baja educación y/o de bajos recursos económicos). A los tres meses (edad corregida), se les observó a las díadas durante la interacción social en el paradigma de Cara a Cara y Rostro Inmutable (FFSF) y durante el juego libre. A los 12 meses, se les observó a las díadas madre-infante por medio de la Situación Extraña de Ainsworth. Se clasificó más de la mitad (58.7%) de los infantes como afectivamente inseguros. Entre los factores de predicción de la afectividad segura se incluyó un patrón de conducta regulatoria con orientación social positiva durante FFSF, una sensibilidad materna más alta y la cooperación del infante durante el juego libre, así como la ausencia de factores de riesgo sociales/familiares. Las variables perinatales no estuvieron relacionadas con la afectividad. Los resultados indican que los factores contextuales, tanto relacionales como sociales contribuyen a la afectividad en este grupo biológicamente vulnerable.

Co-Circulation of Leishmania Parasites and Phleboviruses in a Population of Sand Flies Collected in the South of Portugal.

In the Old World, phlebotomine sand flies from the genus Phlebotomus are implicated in the transmission of Leishmania spp. parasites (Kinetoplastida: Trypanosomatidae) and viruses belonging to the genus Phlebovirus (Bunyavirales: Phenuiviridae). Two of the five sand fly species known to occur in Portugal, Phlebotomus perniciosus and Ph. ariasi, the former being the most ubiquitous, are recognized vectors of Leishmania infantum, which causes visceral leishmaniasis, the most prevalent form of leishmaniasis in the country. Phlebotomus perniciosus is also the vector of the neurotropic Toscana virus, which can cause aseptic meningitis. Entomological surveillance is essential to provide fundamental data about the presence of vectors and the pathogens they can carry. As such, and given the lack of data in Portugal, an entomological survey took place in the Algarve, the southernmost region of the country, from May to October 2018. Polymerase chain reaction assays were performed in order to detect the presence of the above-mentioned pathogens in sand fly pools. Not only were both Leishmania parasites and phleboviruses detected during this study, but more importantly, it was the first time their co-circulation was verified in the same sand fly population collected in Portugal.

(Un)Broken: Lateral violence among hospital nurses, user violence, burnout, and general health: A structural equation modeling analysis.

Introduction: Workplace violence is a social problem yet to be solved. Although it is present in virtually all work environments, its prevalence in healthcare settings stands out, being perceived as something inherent to the job. Most studies in this context have focused on user violence against professionals. However, it has been observed that violence among colleagues in these types of jobs is a risk factor for the health of workers and has rarely been studied as a whole. Among the main consequences of exposure to violence reported in the literature, burnout syndrome, depression, anxiety, or somatic problems have been among the most studied. On the one hand, some authors claim that being exposed to workplace violence can increase the associated physical and psychological pathology and lead to a picture congruent with burnout. On the other hand, it has been hypothesized that violence is associated with burnout, which can trigger physical and psychological symptoms. Taking into account this background, the aim of this study is to explore workplace violence in health personnel, symptomatology, and burnout syndrome through mediation models that allow us to know the interrelationships between the variables. Methods: A cross-sectional design with a double descriptive-associative strategy was used. The sample was composed of 950 nursing professionals from public hospitals. The scales of physical and non-physical violence from users to professionals HABS-U, personal, social, and occupational violence among co-workers using the Health Aggressive Behavior Scale - Co-workers and Superiors (HABS-CS) scale, the burnout scale Maslach Burnout Inventory - General Survey (MBI-GS) which evaluates professional exhaustion, efficacy and cynicism, and the factors referring to depression, anxiety, somatization, and dysfunction of the GHQ-28 scale were applied. In order to calculate the models, workplace violence was used as a predictor of symptomatology, using the burnout variables as mediators. Regression coefficients with and without mediation model, direct and standardized estimates were obtained. For statistical power, Bootstrap analysis was used to calculate direct mediation effects. Results: After controlling the mediation effects of burnout and cynicism, physical and non-physical user violence toward healthcare personnel were significant predictors of the GHQ-28 scores. These same results were obtained when assessing the relationship between social, occupational, and personal violence among co-workers and GHQ-28 scores. Conclusion: Our results contribute to increase the evidence about the effects of violence on the health of professionals and to advance in the characterization of the possible consequent psychological damage. Regardless of the type of violence experienced, exposure to violence can lead to anxious, depressive or somatization symptoms, among others. Violence is also a predictor of burnout syndrome, which in turn accentuates the rest of the consequences studied. Despite the limitations of the proposed model, these results serve to highlight the complexity of the situation experienced by healthcare professionals. Moreover, it serves as a basis for proposing intervention/prevention programs to raise awareness and protect professionals from these risks. To this end, self-care tools should be proposed with which professionals take care of their own health through the management of violent situations and/or the improvement of occupational health.

Sandfly-Borne Phleboviruses in Portugal: Four and Still Counting.

According to ICTV, there are currently 66 known phlebovirus species. More than 40 of these viruses were isolated or detected in phlebotomine sandflies and some of them are known pathogens. In Portugal, information about sandfly-borne phleboviruses is scarce and scattered sandfly-borne diseases are neglected and often not considered in differential diagnoses. The main objective of this work was to gather the existing information and to raise awareness about the circulating phleboviruses in this country. To date, Massilia and Alcube phleboviruses have been isolated from sandflies in southern Portugal. Human infections with Toscana and Sicilian phleboviruses have been reported, as well as seroprevalence in cats and dogs. More studies are needed in order to understand if the viruses isolated during the entomological surveys have an impact on human health and to fully understand the real importance of the already recognized pathogens in our country.

Prosecutorial decision-making regarding offenders' social reintegration programs in intimate partner violence cases. A Portuguese study.

Intimate partner violence is one of the most challenging and demanding problems that the criminal justice system has to face. Given the severe consequences of intimate partner violence, it is imperative that intervention from the criminal justice system, regarding perpetrators, be effective to prevent further victimization and recurrences. In Portugal, it is up to the state prosecutor to decide which cases will be subject to a social reintegration program as a pretrial diversion program. This study aims to explore the variables that might influence the state prosecutor's decision-making process. We have examined 283 intimate partner violence cases in which provisional suspension of criminal proceedings was applied. The decision as to whether defendants should be referred for social reintegration program attendance (G1) or not (G2) was made by the state prosecutor. Differences between G1 and G2 were identified: the victim's age, couple living in a current relationship, drug-addicted defendant, intimate partner violence child exposure. However, defendants' unemployment and drug abuse were the only two variables identified as a determinant for state prosecutor decisions. We believe that the effectiveness of state prosecution decision-making would benefit from: (a) systematically taking into account all intimate partner violence risk factors; (b) an index or checklist detailing what science reveals useful in intimate partner violence offenders' social reintegration; (c) rehabilitation solutions based on the needs of each offender instead of a "one-size-fits-all" approach.

Combined detection of molecular and serological signatures of viral infections: The dual assay concept.

The recent worldwide spread of viral infections has highlighted the need for accurate, fast, and inexpensive disease diagnosis and monitorization methods. Current diagnostics tend to focus either on molecular or serological testing. In this work we propose a dual detection assay approach for viral diseases, where both serological and molecular assays are combined in a single analysis performed on a magnetoresistive system. This type of assay guarantees an accurate assessment of the infection phase, saving time and costs associated with multiple independent tests. Zika and dengue viruses were used as model diseases for the validation of the system. Human IgG anti-zika and anti-dengue antibodies were successfully detected in infected patients' serum, using a novel approach combining competitive and sandwich strategies in a magnetoresistive portable platform. Specificity and sensitivity values of 100% were obtained. Calibration curves with dynamic ranges between 10 ng/mL and 1 µg/mL were established achieving LODs of 1.26 and 1.38 nM for IgG anti-ZIKV and anti-DENV antibodies, respectively. Viral RNA detection down to a few hundreds of pM was also successfully carried out after the design of specific oligo probes and primers for RT-PCR amplification. Dual assays were performed for both viruses, where viral RNA and anti-virus antibodies in serum samples were simultaneously detected. The results obtained for the detection of the molecular and serological targets in the dual assay format show no significant difference between the ones obtained individually, proving the feasibility and accuracy of the dual detection assay. This assay format represents a new paradigm in viral infections diagnostics.

Mutation rate of SARS-CoV-2 and emergence of mutators during experimental evolution.

Background and objectives: To understand how organisms evolve, it is fundamental to study how mutations emerge and establish. Here, we estimated the rate of mutation accumulation of SARS-CoV-2 in vitro and investigated the repeatability of its evolution when facing a new cell type but no immune or drug pressures. Methodology: We performed experimental evolution with two strains of SARS-CoV-2, one carrying the originally described spike protein (CoV-2-D) and another carrying the D614G mutation that has spread worldwide (CoV-2-G). After 15 passages in Vero cells and whole genome sequencing, we characterized the spectrum and rate of the emerging mutations and looked for evidences of selection across the genomes of both strains. Results: From the frequencies of the mutations accumulated, and excluding the genes with signals of selection, we estimate a spontaneous mutation rate of 1.3 × 10 -6 ± 0.2 × 10-6 per-base per-infection cycle (mean across both lineages of SARS-CoV-2 ± 2SEM). We further show that mutation accumulation is larger in the CoV-2-D lineage and heterogeneous along the genome, consistent with the action of positive selection on the spike protein, which accumulated five times more mutations than the corresponding genomic average. We also observe the emergence of mutators in the CoV-2-G background, likely linked to mutations in the RNA-dependent RNA polymerase and/or in the error-correcting exonuclease protein. Conclusions and implications: These results provide valuable information on how spontaneous mutations emerge in SARS-CoV-2 and on how selection can shape its genome toward adaptation to new environments. Lay Summary: Each time a virus replicates inside a cell, errors (mutations) occur. Here, via laboratory propagation in cells originally isolated from the kidney epithelium of African green monkeys, we estimated the rate at which the SARS-CoV-2 virus mutates-an important parameter for understanding how it can evolve within and across humans. We also confirm the potential of its Spike protein to adapt to a new environment and report the emergence of mutators-viral populations where mutations occur at a significantly faster rate.

Intimate partner violence against women. Does violence decrease after the entry of the alleged offender into the criminal justice system?

Intimate partner violence (IPV) is simultaneously assumed as a serious crime and a major public health issue, having recurrences as one of its main characteristics and, consequently, re-entries of some alleged offenders in the criminal justice system (CJS). The main goal of this study is to assess if in cases of female victims of IPV, violence decreases after the first entry of the alleged offender in the CJS. A retrospective study was performed based on the analysis of police reports of alleged cases of IPV during a 4-year period. The final sample (n = 1 488) was divided into two groups according to the number of entries in the CJS (single or multiple) followed by a comparative approach. Results suggest that violence decreases after the first entry of alleged offenders in the CJS. Re-entries were found in only 15.5% of the cases but they were accountable for 3.3 times more crimes on average. Besides, victims of recidivism presented more injuries and required more medical care. Thus, a small group of alleged offenders seems to be more violent and accountable for most of the IPV crimes registered in the CJS suggesting that regardless of legal sanctions aiming to deter violence, these measures may not be enough for a certain group of offenders. This study sustains the need for a predictive model to quantify the risk of repeated IPV cases within the Portuguese population.

Suspension of Criminal Proceedings for Perpetrators of Intimate Partner Violence Against Women: Impact on Re-Entries.

Intimate partner violence (IPV) is one of the most prevalent crimes in our society, but the legal mechanisms to oppose it are recent. The Portuguese Provisional Suspension of Criminal Proceedings (PSCP) as a criminal justice system (CJS) response proposes an integrated consensual solution with the involved parties, to reduce offenders' recidivism. This article analyses the effect of PSCP on re-entries into the CJS. We examined 1,662 IPV police reports, exploring cases that underwent PSCP and re-entries of the same offender in the CJS. Results show that PSCP is applied in 17% of the cases. From all analyzed determinants, with a possible relation to the PSCP implementation, it was found that social violence and the age of both victims and defendants emerge as significantly associated with the request or acceptance of this legal mechanism. No variables tested moderated the relationship between PSCP and re-entry over 96months following the first police report. The article also examined variables that might moderate the decision to request this legal mechanism among victims and defendants.

Tough Love Lessons: Lateral Violence among Hospital Nurses.

BACKGROUND: Workplace violence is a growing social problem among many professions, but it particularly affects the health sector. Studies have mainly focused on evaluating user violence toward health professionals, with less attention being paid to other sources of conflict, such as co-workers themselves. There are different manifestations of this violence in what has been called a context of tolerated or normalized violence among co-workers. However, its effects are far from being tolerable, as they have an impact on general health and job satisfaction and contribute to burnout among professionals. Based on this idea, and following the line of the previous literature, nursing staff are a population at high risk of exposure to workplace violence. For this reason, the present study aims to evaluate exposure to lateral violence or violence among co-workers in nursing staff in public health services and the relationship of this exposure with some of the most studied consequences. (2) Methods: A cross-sectional associative study was carried out in which scales of workplace violence (HABS-CS), burnout (MBI-GS), job satisfaction (OJS), and general health (GHQ-28) were applied to a sample of 950 nursing staff from 13 public hospitals located in the southeast of Spain. (3) Results: The results show that nursing staff have a high exposure to violence from their co-workers, which is more common in male nurses. Greater exposure is observed in professionals with between 6 and 10 years of experience in the profession, and it is not characteristic of our sample to receive greater violence when they have less experience or are younger. A positive correlation is observed with high levels of burnout and a negative correlation with general health and job satisfaction. (4) Conclusions: The results of this work contribute to increasing the scientific evidence of the consequences of a type of workplace violence frequent among nursing staff and to which less attention has been paid in relative terms to other types of prevalent violence. Organizations should be aware of the importance of this type of workplace violence, its frequency and impact, and implement appropriate prevention policies that include the promotion of a culture that does not reward violence or minimize reporting. A change of mentality in the academic environment is also recommended in order to promote a more adequate training of nursing staff in this field.

When Problems Only Get Bigger: The Impact of Adverse Childhood Experience on Adult Health.

Introduction: Previous studies have shown that adverse childhood experiences negatively impact child development, with consequences throughout the lifespan. Some of these consequences include the exacerbation or onset of several pathologies and risk behaviors. Materials and Methods: A convenience sample of 398 individuals aged 20 years or older from the Porto metropolitan area, with quotas, was collected. The evaluation was conducted using an anonymous questionnaire that included sociodemographic questions about exposure to adverse childhood experiences, a list of current health conditions, questions about risk behaviors, the AUDIT-C test, the Fagerström test and the Childhood Trauma Questionnaire-brief form. Variables were quantified to measure adverse childhood experiences, pathologies, and risk behaviors in adult individuals for comparison purposes. Results: Individuals with different forms of adverse childhood experiences present higher rates of smoking dependence, self-harm behaviors, victimization of/aggression toward intimate partners, early onset of sexual life, sexually transmitted infections, multiple sexual partners, abortions, anxiety, depression, diabetes, arthritis, high cholesterol, hypertension, and stroke. Different associations are analyzed and presented. Discussion and Conclusions: The results show that individuals with adverse childhood experiences have higher total scores for more risk behaviors and health conditions than individuals without traumatic backgrounds. These results are relevant for health purposes and indicate the need for further research to promote preventive and protective measures.

Phylogenetic Analysis of Massilia phlebovirus in Portugal.

In the last two decades, molecular surveys of arboviruses have enabled the identification of several new viruses, contributing to the knowledge of viral diversity and providing important epidemiological data regarding possible new emerging viruses. A combination of diagnostic assays, Illumina sequencing and phylogenetic inference are here used to characterize two new Massilia phlebovirus strains isolated from sandflies collected in the Arrábida region, Portugal. Whole genome sequence analysis enabled their identification as reassortants and the recognition of genomic variants co-circulating in Portugal. Much is still unknown about the life cycle, geographic range, evolutionary forces and public health importance of these viruses in Portugal and elsewhere, and more studies are needed.

Long-Term Evolution of SARS-CoV-2 in an Immunocompromised Patient with Non-Hodgkin Lymphoma.

Recent studies have shown that persistent SARS-CoV-2 infections in immunocompromised patients can trigger the accumulation of an unusual high number of mutations with potential relevance at both biological and epidemiological levels. Here, we report a case of an immunocompromised patient (non-Hodgkin lymphoma patient under immunosuppressive therapy) with a persistent SARS-CoV-2 infection (marked by intermittent positivity) over at least 6 months. Viral genome sequencing was performed at days 1, 164, and 171 to evaluate SARS-CoV-2 evolution. Among the 15 single-nucleotide polymorphisms (SNPs) (11 leading to amino acid alterations) and 3 deletions accumulated during this long-term infection, four amino acid changes (V3G, S50L, N87S, and A222V) and two deletions (18-30del and 141-144del) occurred in the virus Spike protein. Although no convalescent plasma therapy was administered, some of the detected mutations have been independently reported in other chronically infected individuals, which supports a scenario of convergent adaptive evolution. This study shows that it is of the utmost relevance to monitor the SARS-CoV-2 evolution in immunocompromised individuals, not only to identify novel potentially adaptive mutations, but also to mitigate the risk of introducing "hyper-evolved" variants in the community. IMPORTANCE Tracking the within-patient evolution of SARS-CoV-2 is key to understanding how this pandemic virus shapes its genome toward immune evasion and survival. In the present study, by monitoring a long-term COVID-19 immunocompromised patient, we observed the concurrent emergence of mutations potentially associated with immune evasion and/or enhanced transmission, mostly targeting the SARS-CoV-2 key host-interacting protein and antigen. These findings show that the frequent oscillation in the immune status in immunocompromised individuals can trigger an accelerated virus evolution, thus consolidating this study model as an accelerated pathway to better understand SARS-CoV-2 adaptive traits and anticipate the emergence of variants of concern.

Toscana Virus: Ten Years of Diagnostics in Portugal.

INTRODUCTION: Toscana virus (TOSV) is an emerging sandfly-borne virus within the Phlebovirus genus. Although most infections caused by this virus present as asymptomatic or with minimal symptomatology, TOSV may emerge as a febrile disease or sporadic cases of neurological disease such as meningitis or meningoencephalitis. This pathogen is distributed throughout the Mediterranean basin, along with the spatial distribution of its recognized sandfly vector, Phlebotomus perniciosus. Portugal, after Italy, was the second country considered endemic for this virus, with the first case of acquired infection published in 1985. Although little is known about the circulation of this virus in Portugal, the laboratory diagnosis of TOSV is available at the Centre for Vectors and Infectious Diseases Research of the National Institute of Health Dr. Ricardo Jorge (CEVDI/INSA), since 2007. The aim of this study is to report the results of the diagnosis of TOSV at the CEVDI/INSA, between 2009 and 2018. MATERIAL AND METHODS: The diagnosis of TOSV in the CEVDI/INSA is included in the arboviruses and vector-borne neurotropic viruses panels or can be performed, when specified, for TOSV only. Direct detection is made in cerebrospinal fluid samples and is available for TOSV by specific real-time reverse transcription polymerase chain reaction followed by conventional real-time reverse transcription polymerase chain reaction for sequencing purposes, if positive. For indirect diagnosis, performed in serum samples, an in-house immunofluorescence assay for the detection of IgM and IgG antibodies against TOSV is used. A commercial immunofluorescence assay consisting in a mosaic of four phleboviruses is also available, in which, in addition to TOSV, antibody detection for sandfly fever Naples virus, sandfly fever Sicilian virus and sandfly fever Cyprus virus can be done. All diagnostic tests requested by clinicians to the CEVDI/INSA for arboviruses, neurotropic viruses and/or TOSV between January 2009 and December 2018, were included in this study. RESULTS: During the study period, the CEVDI/INSA received samples from 608 patients with diagnostic requests for TOSV. Five acute TOSV infections and one acute sandfly fever Sicilian virus infection were confirmed in serum samples. Three other patients had serological evidence of previous contact with the virus. Two of the six patients with acute infection developed febrile syndrome, and the other four presented with neurological disease: meningitis (n = 2), meningoencephalitis (n = 1) and severe depression of consciousness (n = 1). These infections were most likely acquired in the districts of Faro (3), Lisbon (2) and Setúbal (1). DISCUSSION: In Portugal, the number of laboratory diagnostic requests for TOSV is low when compared to the numbers of requests for other less prevalent vector-borne viruses. The Faro district presented the highest number of TOSV-specific diagnostic requests which seems to indicate a higher level of recognition by clinicians in that region. Febrile syndrome and neurological disease were the clinical manifestations that were present in acute cases. In this study, in addition to the Faro district, recent infections were also detected in the districts of Lisbon and Setúbal. It is probable that TOSV may be distributed throughout the mainland territory since its main vector is present from north to south. In 2017, the sandfly fever Sicilian virus was associated for the first time with human disease in our country, thus alerting to the circulation of this phlebovirus. CONCLUSION: Even though the number of identified cases in Portugal is low, TOSV circulates and causes disease in our country. The diagnosis of this and other phleboviruses should not be neglected in the differential diagnosis of febrile syndrome and viral meningitis and meningoencephalitis, especially during the warmer months, when the vector's activity is higher.

Introdução: O vírus Toscana (TOSV) é um vírus emergente, transmitido por flebótomos, que pertence ao género Phlebovirus. Apesar de a maioria das infeções causadas por este vírus serem assintomáticas ou apresentarem uma sintomatologia ligeira, o TOSV pode causar síndrome febril ou casos esporádicos de doença neurológica tal como meningite ou meningoencefalite. Este agente patogénico encontra-se distribuído por toda a bacia do Mediterrâneo, de acordo com as áreas de distribuição do seu vetor reconhecido, Phlebotomus perniciosus. Depois de Itália, Portugal foi o segundo país considerado endémico para este vírus após a publicação, em 1985, do primeiro caso de infeção adquirida no nosso território. Apesar do pouco conhecimento acerca da circulação deste vírus, no nosso país, o diagnóstico laboratorial de TOSV está disponível em Portugal, desde 2007, no Centro de Estudos de Vetores e Doenças Infeciosas do Instituto Nacional de Saúde Dr. Ricardo Jorge (CEVDI/INSA). O objetivo deste trabalho é dar a conhecer os resultados do diagnóstico de TOSV em Portugal, de 2009 a 2018, no CEVDI/INSA. Material e Métodos: O diagnóstico de TOSV no CEVDI/INSA está inserido nos painéis 'arbovírus' e 'vírus neurotrópicos transmitidos por vetores' ou pode ser realizado, quando especificado, só para TOSV. O diagnóstico direto é realizado em amostras de líquido cefalorraquidiano e encontra-se disponível no CEVDI/INSA por RT-PCR em tempo real, específico para TOSV, seguido de RT-PCR convencional, no caso de a amostra ser positiva na primeira técnica, para confirmação por sequenciação. Para o diagnóstico indireto, realizado em amostras de soro, é utilizado uma técnica de imunofluorescência in-house, para a deteção de anticorpos IgM e IgG anti-TOSV. Também está disponível uma imunofluorescência comercial, com um mosaico de quatro flebovírus, onde para além do TOSV, são testados anticorpos contra três vírus da febre por flebótomos, nomeadamente Nápoles, Sicília e Chipre. Neste trabalho foram considerados os pedidos de diagnóstico ao CEVDI/INSA para arbovírus, vírus neurotrópicos e/ou TOSV, de janeiro de 2009 a dezembro de 2018. Resultados: No período em estudo, foram enviadas ao CEVDI/INSA, amostras de 608 indivíduos com pedido de diagnóstico de TOSV. Foram confirmadas cinco infeções agudas por TOSV e uma infeção aguda por vírus Sicília em amostras de soro. Três outros doentes apresentaram prova serológica de contacto prévio com o TOSV. Dois dos doentes com infeção aguda apresentaram síndrome febril, mas quatro evidenciaram quadros neurológicos: meningite (n = 2), meningoencefalite (n = 1) e alterações graves do estado de consciência (n = 1). Estas infeções foram, muito provavelmente, adquiridas nos distritos de Faro (3), Lisboa (2) e Setúbal (1). Discussão: Em Portugal, o número de pedidos de diagnóstico laboratorial para TOSV é baixo quando comparado com o número de pedidos para outros vírus transmitidos por vetores. O distrito de Faro foi o que apresentou o número mais alto de pedidos de diagnóstico específicos para TOSV, o que parece demonstrar que existe um maior reconhecimento pelos clínicos daquela região. Síndrome febril e doença neurológica foram as manifestações clínicas nos casos agudos. Neste estudo, além do distrito de Faro, foram também detetadas infeções recentes nos distritos de Lisboa e Setúbal. É provável que o TOSV se encontre distribuído por todo o território continental, uma vez que o seu principal vetor está presente de norte a sul. Em 2017, o vírus Sicília foi associado, pela primeira vez, a doença humana no nosso país, alertando para a circulação deste flebovírus. Conclusão: Apesar do número de casos identificados em Portugal ser baixo, o TOSV circula e causa doença no nosso país. Este e outros flebovírus não deveriam ser negligenciados no diagnóstico diferencial de síndrome febril e de meningites e meningoencefalites virais, em especial nos meses mais quentes, quando é maior a atividade dos seus vetores.

Congenital SARS-CoV-2 Infection in a Neonate With Severe Acute Respiratory Syndrome.

Coronavirus disease, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is mainly transmitted through droplets, but other ways of transmission have been hypothesized. We report a case of vertical transmission of SARS-CoV-2 in a preterm born to an infected mother, confirmed by the presence of the virus in the neonatal blood, nasopharyngeal and oropharyngeal swabs collected in the first half an hour of life. The neonate presented with acute respiratory distress, similar to the findings in severely affected adults. This case highlights the importance of pregnancy, labor and neonatal period surveillance of affected mothers and their newborns.

Seasonal Dynamics and Spatial Distribution of Aedes albopictus (Diptera: Culicidae) in a Temperate Region in Europe, Southern Portugal.

Aedes albopictus is an invasive mosquito that has colonized several European countries as well as Portugal, where it was detected for the first time in 2017. To increase the knowledge of Ae. albopictus population dynamics, a survey was carried out in the municipality of Loulé, Algarve, a Southern temperate region of Portugal, throughout 2019, with Biogents Sentinel traps (BGS traps) and ovitraps. More than 19,000 eggs and 400 adults were identified from May 9 (week 19) and December 16 (week 50). A positive correlation between the number of females captured in the BGS traps and the number of eggs collected in ovitraps was found. The start of activity of A. albopictus in May corresponded to an average minimum temperature above 13.0 °C and an average maximum temperature of 26.2 °C. The abundance peak of this A. albopictus population was identified from September to November. The positive effect of temperature on the seasonal activity of the adult population observed highlight the importance of climate change in affecting the occurrence, abundance, and distribution patterns of this species. The continuously monitoring activities currently ongoing point to an established population of A. albopictus in Loulé, Algarve, in a dispersion process to other regions of Portugal and raises concern for future outbreaks of mosquito-borne diseases associated with this invasive mosquito species.

Mitogenome diversity of Aedes (Stegomyia) albopictus: Detection of multiple introduction events in Portugal.

Aedes albopictus, along with Ae. aegypti, are key arbovirus vectors that have been expanding their geographic range over the last decades. In 2017, Ae. albopictus was detected for the first time at two distinct locations in Portugal. In order to understand how the Ae. albopictus populations recently introduced in Portugal are genetically related and which is their likely route of invasion, we performed an integrative cytochrome C oxidase I gene (COI)- and mitogenome-based phylogeographic analysis of mosquitoes samples collected in Portugal in 2017 and 2018 in the context of the global Ae. albopictus diversity. COI-based analysis (31 partial sequences obtained from 83 mosquitoes) revealed five haplotypes (1 to 5), with haplotype 1 (which is widely distributed in temperate areas worldwide) being detected in both locations. Haplotypes 2 and 3 were exclusively found in Southern region (Algarve), while haplotype 4 and 5 were only detected in the North of Portugal (Penafiel, Oporto region). Subsequent high discriminatory analyses based on Ae. albopictus mitogenome (17 novel sequences) not only confirmed a high degree of genetic variability within and between populations at both geographic locations (compatible with the Ae. albopictus mosquito populations circulating in Europe), but also revealed two mitogenome mutational signatures not previously reported at worldwide level. While our results generally sustain the occurrence of multiple introduction events, fine mitogenome sequence inspection further indicates a possible Ae. albopictus migration within the country, from the Northern introduction locality to the Southern region. In summary, the observed scenario of high Ae. albopictus genetic diversity in Portugal, together with the detection of mosquitoes in successive years since 2017 in Algarve and Penafiel, points that both Ae. albopictus populations seem to be already locally established, as its presence has been reported for three consecutive years, raising the public health awareness for future mosquito-borne diseases outbreaks.

Maternal pre and perinatal experiences with their full-term, preterm and very preterm newborns.

BACKGROUND: Mothers' reports about pregnancy, maternity and their experiences during the perinatal period have been associated with infants' later quality of attachment and development. Yet, there has been little research with mothers of very preterm newborns. This study aimed to explore mothers' experiences related to pregnancy, premature birth, relationship with the newborn, and future perspectives, and to compare them in the context of distinct infants' at-birth-risk conditions. METHODS: A semi-structured interview was conducted with women after birth, within the first 72 h of the newborn's life. A total of 150 women participated and were divided in three groups: (1) 50 mothers of full-term newborns (Gestational Age (GA) ≥ 37 weeks; FT), (2) 50 mothers of preterm newborns (GA 32-36 weeks; PT) and (3) 50 mothers of very preterm newborns (GA < 32 weeks; VPT). RESULTS: Mothers of full-term infants responded more often that their children were calm and that they did not expect difficulties in taking care of and providing for the baby. Mothers of preterm newborns although having planned and accepted well the pregnancy (with no mixed or ambivalent feelings about it) and while being optimistic about their competence to take care of the baby, mentioned feeling frightened because of the unexpected occurrence of a premature birth and its associated risks. Mothers of very preterm newborns reported more negative and distressful feelings while showing more difficulties in anticipating the experience of caring for their babies. CONCLUSION: The results indicate that Health Care Systems and Neonatal Care Policy should provide differentiated psychological support and responses to mothers, babies and families, taking into account the newborns' GA and neonatal risk factors.

Abundance and Updated Distribution of Aedes aegypti (Diptera: Culicidae) in Cabo Verde Archipelago: A Neglected Threat to Public Health.

BACKGROUND: Mosquito-borne viruses, such as Zika, dengue, yellow fever, and chikungunya, are important causes of human diseases nearly worldwide. The greatest health risk for arboviral disease outbreaks is the presence of the most competent and highly invasive domestic mosquito, Aedes aegypti. In Cabo Verde, two recent arbovirus outbreaks were reported, a dengue outbreak in 2009, followed by a Zika outbreak in 2015. This study is the first entomological survey for Ae. aegypti that includes all islands of Cabo Verde archipelago, in which we aim to evaluate the actual risk of vector-borne arboviruses as a continuous update of the geographical distribution of this species. METHODS: In order to assess its current distribution and abundance, we undertook a mosquito larval survey in the nine inhabited islands of Cabo Verde from November 2018 to May 2019. Entomological larval survey indices were calculated, and the abundance analyzed. We collected and identified 4045 Ae. aegypti mosquitoes from 264 positive breeding sites in 22 municipalities and confirmed the presence of Ae. aegypti in every inhabited island. Results: Water drums were found to be the most prevalent containers (n = 3843; 62.9%), but puddles (n = 27; 0.4%) were the most productive habitats found. The overall average of the House, Container, and Breteau larval indices were 8.4%, 4.4%, and 10.9, respectively. However, 15 out of the 22 municipalities showed that the Breteau Index was above the epidemic risk threshold. CONCLUSION: These results suggest that if no vector control measures are considered to be in place, the risk of new arboviral outbreaks in Cabo Verde is high. The vector control strategy adopted must include measures of public health directed to domestic water storage and management.

Simultaneous extraction and purification of fucoxanthin from Tisochrysis lutea microalgae using compressed fluids.

The marine microalga Tisochrysis lutea, a Haptophyta with a thin cell wall and currently used mainly in aquaculture is a potential source of several bioactive compounds of interest such as carotenoids. In the present study, the simultaneous extraction and purification of fucoxanthin, the main carotenoid from T. lutea, was optimized using pressurized fluid extraction followed by in-cell purification. An experimental design was employed to maximize carotenoids' extraction; the experimental factors chosen were: (i) percentage of ethanol/ethyl acetate (0-100 %), (ii) temperature (40-150°C), and (iii) number of static extraction cycles (1-3). The maximum carotenoids' recovery, mainly fucoxanthin, was obtained with pure ethyl acetate at 40°C using one extraction cycle, achieving values of 132.8 mg of carotenoids per gram of extract. Once the optimum extraction conditions were confirmed, in-cell purification strategies using different adsorbents were developed to obtain fucoxanthin-enriched extracts. Activated charcoal showed potential retention of chlorophylls allowing an effective purification of fucoxanthin in the obtained extracts. Chemical characterization of extracts was carried out by reversed-phase high-performance liquid chromatography with diode array detection. Therefore, a selective fractionation of high value compounds was achieved using the proposed green downstream platform based on the use of compressed fluids.