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Extracellular vesicles (EVs) are membrane-bound structures released by cells and tissues into biofluids, involved in cell-cell communication. In humans, circulating red blood cells (RBCs), represent the most common cell-type in the body, generating daily large numbers of microvesicles. In vitro, RBC vesiculation can be mimicked by stimulating RBCs with calcium ionophores, such as ionomycin and A23187. The fate of microvesicles released during in vivo aging of RBCs and their interactions with circulating cells is hitherto unknown. Using SEC plus DEG isolation methods, we have found that human RBCs generate microvesicles with two distinct sizes, densities, and protein composition, identified by flow cytometry, and MRPS, and further validated by immune TEM. Furthermore, proteomic analysis revealed that RBC-derived microvesicles (RBC-MVs) are enriched in proteins with important functions in ion channel regulation, calcium homeostasis, and vesicular transport, such as of sorcin, stomatin, annexin A7, and RAB proteins. Cryo-electron microscopy identified two separate pathways of RBC-MV-neutrophil interaction, direct fusion with the plasma membrane and internalization, respectively. Functionally, RBC-MVs decrease neutrophil ability to phagocytose E. coli but do not affect their survival at 24 hrs. This work brings new insights regarding the complexity of the RBC-MVs biogenesis, as well as their possible role in circulation., Competing Interests: conflict-of-interest statements CCP is a senior translational data scientist at Sanofi and declares no competing financial interests. The remaining authors declare no competing financial interests that are relevant to the content of this article.

In 2015, Brazil reported an outbreak identified as Zika virus (ZIKV) infection associated with congenital abnormalities. To date, a total of 86 countries and territories have described evidence of Zika infection and recently the appearance of the African ZIKV lineage in Brazil highlights the risk of a new epidemic. The spectrum of ZIKV infection-induced alterations at both cellular and molecular levels is not completely elucidated. Here, we present for the first time the gene expression responses associated with prenatal ZIKV infection from ocular cells. We applied a recently developed non-invasive method (impression cytology) which use eye cells as a model for ZIKV studies. The ocular profiling revealed significant differences between exposed and control groups, as well as a different pattern in ocular transcripts from Congenital Zika Syndrome (CZS) compared to ZIKV-exposed but asymptomatic infants. Our data showed pathways related to mismatch repair, cancer, and PI3K/AKT/mTOR signaling and genes probably causative or protective in the modulation of ZIKV infection. Ocular cells revealed the effects of ZIKV infection on primordial neuronal cell genes, evidenced by changes in genes associated with embryonic cells. The changes in gene expression support an association with the gestational period of the infection and provide evidence for the resulting clinical and ophthalmological pathologies. Additionally, the findings of cell death- and cancer-associated deregulated genes raise concerns about the early onset of other potential pathologies including the need for tumor surveillance. Our results thus provide direct evidence that infants exposed prenatally to the Zika virus, not only with CZS but also without clinical signs (asymptomatic) express cellular and molecular changes with potential clinical implications., (Copyright © 2022 Elsevier Inc. All rights reserved.)

This study aimed to problematize aspects of the work of professors in relation to gender, assessment policies, and health. For this purpose, a qualitative social investigation was conducted within the aspect of participatory studies and the views of materialist feminism. An analysis of the material was carried out through content analysis, according to the topic, using four main themes: conflicts between professors' work and domestic work; professors' work, motherhood, and guilt; policies for the assessment of professors and gender relations; and the sexual division of labor and teaching. It was possible to perceive just how much the demands of the productive sphere have gone beyond the time of the workday and into the reproductive sphere and the private life of professors, compromising the struggle for health and leading to processes of suffering and illness. The theme of the sexual division of labor in public universities appears to be an important issue that highlights work overload and psychological illness, especially at a time when the teaching profession is becoming increasingly competitive. In conclusion, we believe that there is an imperative need for investments in public policies that can guarantee gender equality in higher education.

MicroRNAs are short, non-coding RNA sequences involved in gene expression regulation. Quantification of miRNAs in biological fluids involves time consuming and laborious methods such as Northern blotting or PCR-based techniques. Molecular beacons (MB) are an attractive means for rapid detection of miRNAs, although the need for sophisticated readout methods limits their use in research and clinical settings. Here, we introduce a novel method based on delayed electrophoretic mobility, as a quantitative means for detection of miRNAs-MB hybridization. Upon hybridization with the target miRNAs, MB form a fluorescent duplex with reduced electrophoretic mobility, thus bypassing the need for additional staining. In addition to emission of light, the location of the fluorescent band on the gel acts as an orthogonal validation of the target identity, further confirming the specificity of binding. The limit of detection of this approach is approximately 100 pM, depending on the MB sequence. The method is sensitive enough to detect specific red blood cell miRNAs molecules in total RNA, with single nucleotide specificity. Altogether, we describe a rapid and affordable method that offers sensitive detection of single-stranded small DNA and RNA sequences., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

In 2015, ZIKV infection attracted international attention during an epidemic in the Americas, when neurological disorders were reported in infants who had their mothers exposed to ZIKV during pregnancy. World Health Organization (WHO) epidemiological data show that 5 to 15% of neonates exposed to ZIKV in the uterus have complications included in abnormalities related to Congenital Zika Syndrome (CZS). The risk of complications after birth is not well documented, however, clinical evidence shows that 6% of infants exposed to ZIKV during pregnancy have complications present at birth, and this rate rises to 14% when medical monitoring is performed in all exposed infants, regardless of birth condition. Thus, the evaluation and monitoring of all exposed infants are of foremost importance as the development of late complications has been increasingly supported by clinical evidence. The identification of changes in protein profile of infants exposed to ZIKV without CZS could provide valuable findings to better understand molecular changes in this cohort. Here, we use a shotgun-proteomics approach to investigate alterations in the serum of infants without CZS symptoms but exposed to intrauterine ZIKV (ZIKV) compared to unexposed controls (CTRL). A complex pattern of differentially expressed proteins was identified, highlighting the dysregulation of proteins involved in axon orientation, visual phototransduction, and global protease activity in children exposed to ZIKV without CZS. These data support the importance of monitoring children exposed to ZIKV during gestation and without early CZS symptoms. Our study is the first to assess molecular evidence of possible late disorders in children victims of the ZIKV outbreak in the Americas. We emphasize the importance of medical monitoring of symptomatic and asymptomatic children, as apparently unexplained late neurological and eye disorders may be due to intrauterine ZIKV exposure., (Copyright © 2020 Macedo-da-Silva, Rosa-Fernandes, Barbosa, Angeli, Carvalho, de Oliveira Vianna, Carvalho, Larsen, Cardoso and Palmisano.)

Congenital Zika syndrome was first described due to increased incidence of congenital abnormalities associated with Zika virus (ZIKV) infection. Since the eye develops as part of the embryo central nervous system (CNS) structure, it becomes a specialized compartment able to display symptoms of neurodegenerative diseases and has been proposed as a noninvasive approach to the early diagnosis of neurological diseases. Ocular lesions result from defects that occurred during embryogenesis and can become apparent in newborns exposed to ZIKV. Furthermore, the absence of microcephaly cannot exclude the occurrence of ocular lesions and other CNS manifestations. Considering the need for surveillance of newborns and infants with possible congenital exposure, we developed a method termed cellular imprinting proteomic assay (CImPA) to evaluate the ocular surface proteome specific to infants exposed to ZIKV during gestation compared to nonexposure. CImPA combines surface cells and fluid capture using membrane disks and a large-scale quantitative proteomics approach, which allowed the first-time report of molecular alterations such as neutrophil degranulation, cell death signaling, ocular and neurological pathways, which are associated with ZIKV infection with and without the development of congenital Zika syndrome, CZS. Particularly, infants exposed to ZIKV during gestation and without early clinical symptoms could be detected using the CImPA method. Lastly, this methodology has broad applicability as it could be translated in the study of several neurological diseases to identify novel diagnostic biomarkers. Data are available via ProteomeXchange with identifier PXD014038.

Importance: Non-invasive techniques for retrieving ocular surface cells from babies infected by zika virus (ZIKV) during the gestational period remain to be determined., Objectives: The aim of this study was to describe an optimized impression cytology method for the isolation of viable cells from Zika infected babies with and without Congenital Zika Syndrome (CZS) in satisfactory amount and quality to enable easy adoption in the field and application in the context of genomic and molecular approaches., Design Settings and Participants: Ocular surface samples were obtained with a hydrophilic nitrocellulose membrane (through optimized impression cytology method) from twelve babies referred to the Pediatric Service of the Antonio Pedro Hospital, Universidade Federal Fluminense (UFF), Niteroi, Rio de Janeiro, Brazil. After an authorized written informed consent from the parents, samples were collected from both eyes of 12 babies (4 babies with maternal ZIKV exposure during gestation and presence of clinical signs which included ocular abnormalities and microcephaly; 4 babies with maternal ZIKV exposure during gestation but no clinical signs; and 4 unaffected control babies with negative PCR for Zika virus and without clinical signs). Cells were used for microscopy analyses and evaluated for their suitability for downstream molecular applications in transcriptomic and proteomic experiments., Results: Our optimized impression cytology protocol enabled the capture of a considerable number of viable cells. The microscopic features of the conjunctival epithelial cells were described by both direct analysis of the membrane-attached cells and analysis of cytospinned captured cells using several staining procedures. Epithelial basal, polyhedral and goblet cells were clearly identified in all groups. All cases of ZIKV infected babies showed potential morphological alterations (cell keratinization, pyknosis, karyolysis, anucleation, and vacuolization). Molecular approaches were also performed in parallel. Genomic DNA and RNA were successfully isolated from all samples to enable the establishment of transcriptomic and proteomic studies., Conclusions and Relevance: Our method proved to be a suitable, fast, and non-invasive tool to obtain ocular cell preparations from babies with and without Zika infection. The method yielded sufficient cells for detailed morphological and molecular analyses of samples. We discuss perspectives for the application of impression cytology in the context of ZIKV studies in basic and clinical research., (Copyright © 2019 Barbosa, dos Santos, Silva, Rosa-Fernandes, Pinto, Spínola, Bonvicino, Fernandes, Lucena, Palmisano, Melo, Cardoso and Lemos.)

The number of Tetrastischus howardi (Olliff) females to be released and their dispersion should be known, that way, used D. saccharalis pupae as sentinel host to measure parasitism as function of the release density of the parasitoid and its location in the field. Two sets of trials were run aiming first to define the number of parasitoids to be released and the dispersal of the parasitoid using plots in sugarcane commercial fields, respectively. Pieces of sugarcane stalk holding sentinel pupae were taken to the field and exposed to parasitism in both trials. The parasitoid was released at the rate of 20, 40, 80, and 160 females per sentinel pupa, except for the control plot without releasing. The parasitism rate was calculated based on the recovered pupae after 96 h of exposure time from releasing the parasitoids. The models estimated the best parasitism rate by releasing 102 parasitoids per pupa. In the second trial, sentinel pupae were arranged in five subsequent circles corresponding 4, 8, 12, 16, and 20 m around the central parasitoid releasing point at rate of 4, 8, 12, 16, and 18 pupae per circle. The mean estimated dispersal distance was 7.64 m, with a covering area of 80.07 m 2 . Based on these findings, release of T. howardi is recommended in 125 points per hectare of sugarcane at rate of 102 females per pupa of the pest aiming to achieve homogeneous distribution and parasitism.

The main objective of this article is to present and analyze the research instrument called "health and work notebooks", focusing on the production of knowledge concerning professors' work at a public university. The notebooks serve as a qualitative and participant research technique that is appropriate for the in-depth study of relations between health and work, viewing workplaces as privileged spaces for exercising effective action in the defense of health and the work experience as the principal material for analysis. The notebooks' special quality as a research technique lies in worker's role as protagonist in the research, as the diary's author and co-participant in the study. Eight professors participated, all from the same institute in a federal university (IFES) in Rio de Janeiro, Brazil. As for analysis of the empirical materials from the notebooks and consistent with the workers, the thematic analysis technique was adopted, producing four main discussion categories: time on the job and professors' multiple work activities; precarization of working conditions at universities; faculty health at limits; and the notebooks viewed from the authors' perspective. As for the results, the theme that stood out was work overload and time pressure to meet targets. Finally, the health and work notebooks proved to be a potential research tool for generating knowledge from a collective perspective.

The article discusses drug abuse and eating disorders from the critical gender and healthcare perspectives, postulating that subjective suffering can be expressed in the body through psychosomatic illnesses. From this perspective, craving for drugs or superfluous consumer goods, just as illness from self-imposed hunger in pursuit of an ideal of slimness, as in anorexia and bulimia, can be symptoms that expose the woman's suffering. A review in the fields of public health and feminist theories highlights the magnitude of the phenomena of medicalization and commodification of health in the psychiatrization of female discomfort. In the gender transition in capitalist societies, social demands for the performance of old and new women's roles accentuate feelings of inadequacy, expressed as the gender discomfort permeating drug abuse and eating disorders, analyzed as diseases of protest. The study proposes to reclaim the ideals of the Program for Comprehensive Women's Healthcare to deal with such challenges.

Visceral motility dysfunction is a key feature of genetic disorders such as megacystis-microcolon-intestinal hypoperistalsis syndrome (MMIHS, MIM moved from 249210 to 155310), chronic intestinal pseudo-obstruction (CIPO, MIM609629), and multisystemic smooth muscle dysfunction syndrome (MSMDS, MIM613834). The genetic bases of these conditions recently begun to be clarified with the identification of pathogenic variants in ACTG2, ACTA2, and MYH11 in individuals with visceral motility dysfunction. The MMIHS was associated with the heterozygous variant in ACTG2 and homozygous variant in MYH11, while the heterozygous variant in ACTA2 was observed in patients with MSMDS. In this study, we describe the clinical data as well as the molecular investigation of seven individuals with visceral myopathy phenotypes. Five patients presented with MMIHS, including two siblings from consanguineous parents, one had CIPO, and the other had MSMDS. In three individuals with MMIHS and in one with CIPO we identified heterozygous variant in ACTG2, one being a novel variant (c.584C>T-p.Thr195Ile). In the individual with MSMDS we identified a heterozygous variant in ACTA2. We performed the whole-exome sequencing in one sibling with MMIHS and her parents; however, the pathogenic variant responsible for her phenotype could not be identified. These results reinforce the clinical and genetic heterogeneity of the visceral myopathies. Although many cases of MMIHS are associated with ACTG2 variants, we suggest that other genes, besides MYH11, could cause the MMIHS with autosomal recessive pattern. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

The oral rehabilitation of edentulous patients with immediate loading has become a safe procedure with high predictability. The success is related to immediate fabrication of a passive fit framework to attach the implants. Based on these considerations, this case report shows an alternative technique for mandibular rehabilitation using implants immediately loaded, where the framework was fabricated using cylinders with internal reinforcement and precast pieces, electrowelding, and conventional welding providing esthetics and function to the patient in a short period of time.

Understanding mortality patterns and interactions between entomopathogenic fungi and parasitoids is important to improve insect biological control programs. The aim of this study was to evaluate the effect of Metarhizium anisopliae (Metschnikoff, 1879) Sorokin, 1833 and Beauveria bassiana (Balsamo) Vuillemin, 1912 (Hypocreales: Clavicipitaceae) on adults of Cotesia flavipes (Cameron, 1891) (Hymenoptera: Braconidae) with biological insecticides Biometha WP Plus (M. anisopliae), Biovéria G (B. bassiana), Boverril WP (B. bassiana), Metarril WP (M. anisopliae), and Metie WP (M. anisopliae) at concentrations of 1 x 10(9) conidia (con).mL(-1), 5 x 10(9) con.ml(-1), and 10 x 10(9) con.ml(-1). In the experimental, 10 females of C. flavipes were packed in disposable cups capped with a contact surface (filter paper, 9 cm2) treated with commercial product. The experimental design was completely randomized, with 16 treatments and five replicates of 10 females each. Mortality was assessed at 24, 48, 72, 96, and 120 hours after exposition (HAE) of the products. In general, B. bassiana and M. anisopliae in the concentrations of 1 x 10(9) con.ml(-1), 5 x 10(9) con.ml(-1), and 10 x 10(9) con.ml(-1) can't affect C. flavipes females because the peak of mortality in treatments with bioinsecticides was similar to the control and this demonstrated the selectivity of fungi B. bassiana and M. anisopliae on C. flavipes females.

Purpose: To identify constitutional alterations of the retinoblastoma 1 gene (RB1) in two cohorts of Brazilian patients with retinoblastoma and to analyze genotype-phenotype associations., Methods: Molecular screening was carried out by direct sequencing of the 27 RB1 exons and flanking regions in blood DNA of 71 patients with retinoblastoma and 4 relatives with retinoma, and with multiplex ligation-dependent probe amplification (MLPA) in 21 patients. The presumed impact of nucleotide substitutions on the structure of the retinoblastoma protein (pRB) was predicted by Polymorphism Phenotyping-2 (PolyPhen-2). Kaplan-Meier and log-rank test were used for estimating 60-month survival rates., Results: One hundred two nucleotide substitutions were detected, 92 substitutions in 59 patients with retinoblastoma and 10 substitutions in 4 individuals with retinoma. Eight substitutions were novel. The majority of substitutions were intronic (86.2%). More than one substitution was present in 37.3% of patients. Twenty-one duplications and 11 deletions were found in 12 patients; some of which with both types of alterations. Duplications/deletions were found in four patients lacking constitutional alterations when analyzed by sequencing, and in eight patients carrying one or more polymorphic intronic substitutions. The global 60-month survival rate in patients was 91.8% (Confidence Interval95% = 85.0 - 99.1). Significant, lower survival rates were found in extraocular presentation (81.0%) versus intraocular tumors (P = 0.014), first enucleation after 1 month following diagnosis (80.9%) versus earlier first enucleation (P = 0.020), and relapse (100.0%) versus absence of relapse (P = 0.0005)., Conclusions: Fifteen substitutions (4 intronic and 11 exonic) were identified as probably or likely pathogenic. Four of these 11 exonic substitutions were novel. Survival rates, however, were not affected by presence of these probably or likely pathogenic alterations, most of which not found in patients with retinoblastoma from other Latin American countries. These differences might be related to the different ethnic composition of the Latin American cohorts. Portuguese Abstract.

Background: The purpose of this study was to identify mutations associated with bilateral retinoblastoma in a quadruplet conceived by in vitro fertilization, and to trace the parental origin of mutations in the four quadruplets and their father., Methods: Mutational screening was carried out by sequencing. Genotyping was carried out for determining quadruplet zygosity., Results: The proband was a carrier of a novel RB1 constitutive mutation (g.2056C>G) which was not detected in her father or her unaffected sisters, and of two other mutations (g.39606 C>T and g.174351T>A) also present in two monozygotic sisters. The novel mutation probably occurred de novo while the others were of likely maternal origin. The novel mutation, affecting the Kozak consensus at the 5'UTR of RB1 and g.174351T>A were likely associated to retinoblastoma in the proband., Conclusion: Molecular diagnosis of retinoblastoma requires genotypic data of the family for determining hereditary transmission. In the case of children generated by IVF with oocytes from an anonymous donor which had been stored in a cell repository, this might not be successfully accomplished, making precise diagnosis impracticable for genetic counseling.

Based on an extensive review of specialized literature about woman smoking, this essay aims to promote a better understanding of this issue, proposing the adoption of Social Sciences concepts, particularly at gender category, to support more comprehensive and encompassing approaches towards prevention and health assistance of tobacco smoking women. Analyzing the epidemiologic scenario of woman smoking, three tendencies could be identified--pauperization, feminilization and juvenilization--confirming that many of women disease are related to social and gender inequalities. Gender dimension is associated to woman smoking through women's protest pathologies which historically express dissatisfactions and social contradictions experienced by women. The essay concludes that the meaning attributed to cigarette by women has strong connections with the ways gender relations are organized in current society, as well as with their relationships with health services, demanding broader and integral approaches of women's health, including woman smoking.

This article presents the main results of a study that examined the symbolic and material meanings of women's smoking, adopting a critical and qualitative gender approach. Semi-structured interviews were held with 14 women smokers in different stages of the smoking cessation process. The research locus was a tobacco treatment program located in a public hospital in Rio de Janeiro, Brazil. The findings showed how deeply smoking is interwoven in these women's social and gender trajectories, playing a decisive support role when they have to deal with various difficulties in life. The cigarette as a "companion" emerged as the main empirical category, as something always available to quell anxiety and loneliness, as well as a source of pleasure and relaxation. The critical gender approach evidenced how women's reproductive and productive work overload reinforces their tobacco dependency. In health care, in order to attain women smokers' adherence to the arduous cessation process, it is crucial to consider the complex relations between social and gender dimensions when cigarettes are viewed as a "companion".

This article presents the results of research in 2003 focusing on the relationship between health care professionals from a public clinic and HIV positive women, adopting a gender approach. Ten patients and seven attending professionals were interviewed on the following themes: gender representations; representations of HIV/AIDS in women; and accommodation and resistance strategies to internalized attitudes and values regarding the health professional/patient relationship. According to the results, health professionals attempted to adjust the treatment to the limited resources of both the institution and the patient, endeavoring to offer the most complete care possible. Their social imagery continues to portray the traditional role of woman/mother/wife, perceived as a victim. Meanwhile patients adopted a "fighting woman" image, reinforced by the "modern woman" ideology, produced in both public and private spheres, disguising the effects of the women's double workload. Contrary to the myth of passivity, patients took an active stance towards their treatment, negotiating with health professionals about their own needs and objective possibilities. Despite the material and symbolic limitations, the potential was identified for changes in health care in order to achieve full reproductive rights.

This study approaches the social and cultural profile concerning risk for HIV infection in women, describing some epidemiological variables and disclosing reports of risk situations, the meaning of living with AIDS, and support received. A semi-structured questionnaire was used to interview 25 women from the University Hospital of the Federal University of Rio de Janeiro, prior to the availability of multi-drug treatment. The majority reported limited schooling, were housewives or engaged in under-skilled occupations, and had family incomes lower than average for users of this public teaching hospital. The view of AIDS as "someone else's disease" was prevalent, and STDs were perceived as male infections, although several women reported episodes of STDs prior to HIV. They had received their diagnosis and initial medical care only after their partners' and/or children's illness or death. The study points to preventive strategies reinforcing these silent women's bargaining power, acting on men as potential active participants in reproductive health programs that incorporate STD/AIDS issues.