Immunogenic Treatment of Metastatic Breast Cancer Using Targeted Carbon Nanotube Mediated Photothermal Therapy in Combination with Anti-Programmed Cell Death Protein-1.

The high prevalence of breast cancer is a global health concern, compounded by the lack of safe or effective treatments for its advanced stages. These facts urge the development of novel treatment strategies. Annexin A5 (ANXA5) is a natural human protein that binds with high specificity to phosphatidylserine, a phospholipid tightly maintained in the inner leaflet of the cell membrane on most healthy cells but externalized in tumor cells and the tumor vasculature. Here, we have developed a targeted photosensitizer for photothermal therapy (PTT) of solid tumors through the functionalization of single-walled carbon nanotubes (SWCNTs) to ANXA5-the SWCNT-ANXA5 conjugate. The ablation of tumors through the SWCNT-ANXA5-mediated PTT synergizes with checkpoint inhibition, creating a systemic anticancer immune response. In vitro ablation of cells incubated with the conjugate promoted cell death in a dose-dependent and targeted manner. This treatment strategy was tested in vivo with the orthotopic EMT6 breast tumor model in female balb/cJ mice. Enhanced therapeutic effects were achieved by using intratumoral injection of the conjugate and treating tumors at a lower PTT temperature (45°C). Intratumoral injection prevented the accumulation of the SWCNTs in major clearance organs. When combined with checkpoint inhibition of anti-programmed cell death protein-1, SWCNT-ANXA5-mediated PTT increased survival and 80% of the mice survived for 100 days. Evidence of immune system activation by flow cytometry of splenic cells strengthens the hypothesis of an abscopal effect as a mechanism of prolonged survival. SIGNIFICANCE STATEMENT: This study demonstrated a relatively high survival rate (80% at 100 days) of mice with aggressive breast cancer when treated with photothermal therapy using the SWCNT-ANXA5 conjugate injected intratumorally and combined with immune stimulation using the anti-programmed cell death protein-1 checkpoint inhibitor. Photothermal therapy was accomplished by maintaining the tumor temperature at a relatively low level of 45°C and avoiding accumulation of the nanotubes in the clearance organs by using intratumoral administration.

The most common inadequacies in red blood cell requests at a reference center in Western Paraná state.

INTRODUCTION: In the current scenario, in which evidence-based medicine is almost obligatory, therapeutic approaches are increasingly being restricted to measures that are proven to bring benefits to the patient. Transfusion therapy follows the same concept. Even though it can be an essential support procedure in some situations, it must be carefully used, exclusively in situations in which the literature assures scientific support for its usage. Transfusion exposes patients to risks and complications, so it is necessary that we, as doctors, continue working to reduce possible mistakes involving the practice of this type of therapy. OBJECTIVE: To analyze if the red blood cell requests are properly made, analyzing the indication written on them (by the doctor, comparing them to the references in the actual literature. METHOD: In this research, we have analyzed and reviewed the requests for red blood cell concentrates between August and September of 2018 received at the Hematology and Hemotherapy Center studied to find the most common mistakes made by requesters/doctors. RESULTS: A total of 397 of the 754 analyzed requests were evaluated as inadequate. Therefore, 1 out of every 2 transfusions performed did not have their requests properly completed in the city of Cascavel, Paraná. CONCLUSIONS: The number of inappropriate requests was high during the studied period, especially when compared to studies available in the literature. The number of inadequacies suggests that the blood component requests and the subsequent evaluation by the transfusion agency needs to be better addressed in clinical practice.

The most common inadequacies in red blood cell requests at a reference center in Western Paraná state

Abstract Introduction In the current scenario, in which evidence-based medicine is almost obligatory, therapeutic approaches are increasingly being restricted to measures that are proven to bring benefits to the patient. Transfusion therapy follows the same concept. Even though it can be an essential support procedure in some situations, it must be carefully used, exclusively in situations in which the literature assures scientific support for its usage. Transfusion exposes patients to risks and complications, so it is necessary that we, as doctors, continue working to reduce possible mistakes involving the practice of this type of therapy. Objective To analyze if the red blood cell requests are properly made, analyzing the indication written on them (by the doctor, comparing them to the references in the actual literature. Method In this research, we have analyzed and reviewed the requests for red blood cell concentrates between August and September of 2018 received at the Hematology and Hemotherapy Center studied to find the most common mistakes made by requesters/doctors. Results A total of 397 of the 754 analyzed requests were evaluated as inadequate. Therefore, 1 out of every 2 transfusions performed did not have their requests properly completed in the city of Cascavel, Paraná. Conclusions The number of inappropriate requests was high during the studied period, especially when compared to studies available in the literature. The number of inadequacies suggests that the blood component requests and the subsequent evaluation by the transfusion agency needs to be better addressed in clinical practice.

Fetal hemoglobin-boosting haplotypes of BCL11A gene and HBS1L-MYB intergenic region in the prediction of clinical and hematological outcomes in a cohort of children with sickle cell anemia.

Single nucleotide polymorphisms (SNPs) of BCL11A gene and HBS1L-MYB intergenic region (named HMIP-2) affect both fetal hemoglobin (HbF) concentration and clinical outcomes in patients with sickle cell anemia (SCA). However, no previous study has examined the interaction among these SNPs in the regulation of HbF. We examined whether HbF-boosting haplotypes combining alleles of functional SNPs of BCL11A and HMIP-2 were associated with clinical outcomes and hematological parameters, and whether they interact to regulate HbF in a cohort of Brazilian children with SCA. The minor haplotype of BCL11A ("TCA", an allele combination of rs1427407, rs766432, and rs4671393) was associated with higher HbF, hemoglobin and lower reticulocytes count compared to reference haplotype "GAG". The minor haplotype of HMIP-2 ("CGC", an allele combination of rs9399137, rs4895441, and rs9494145) was associated with higher HbF and hemoglobin compared to reference haplotype "TAT". Subjects carrying minor haplotypes showed reduced rate of clinical complications compared to reference haplotypes. Non-carriers of both minor haplotypes for BCL11A and HMIP-2 showed the lowest HbF concentration. Subjects carrying only the minor haplotype of BCL11A showed significantly higher HbF concentration than non-carriers of any minor haplotype, which showed no significant difference compared to subjects carrying only the minor haplotype of HMIP-2. Interestingly, subjects carrying both minor haplotypes of BCL11A ("TCA") and HMIP-2 ("CGC") showed significantly higher HbF levels than subjects carrying only the minor haplotype of BCL11A. Our novel findings suggest that HbF-boosting haplotypes of BCL11A and HMIP-2 can predict clinical outcomes and may interact to regulate HbF in patients with SCA.

Annexin A5 as a targeting agent for cancer treatment.

Identifying a universal biomarker for cancer treatment remains a major challenge in cancer therapy. Extracellular exposure of phosphatidylserine (PS) is tightly regulated and is an "eat me" signal for phagocytosis in healthy cells. Although cancer cells and vasculature express high levels of externalized PS, they do not undergo apoptosis, making them a promising biomarker for cancer treatment. Annexin A5 (ANXA5) is the native binding partner of PS and can actively target and deliver chemotherapies to the tumor microenvironment (TME) via PS expression. ANXA5 acts as a bridge between the innate and adaptive immune systems and contributes to an immunostimulatory profile in the TME. ANXA5-enzyme prodrug therapies allow for systemic delivery of prodrugs and targeted killing at the tumor site. ANXA5-carbon nanotube conjugates have been used to physically ablate tumors via photothermal therapy. This review aims to explore the expression of PS in cancer cells and how ANXA5 has been used as a chemotherapeutic and targeting agent for cancer.

Targeted Single-Walled Carbon Nanotubes for Photothermal Therapy Combined with Immune Checkpoint Inhibition for the Treatment of Metastatic Breast Cancer.

The greatest contributors to cancer mortality are metastasis and the consequences of its treatment. Here, we present a novel treatment of metastatic breast cancer that combines photothermal therapy with targeted single-walled carbon nanotubes (SWCNTs) and immunostimulation with a checkpoint inhibitor. We find that the selective near-infrared photothermal ablation of primary orthotopic EMT6 breast tumors in syngeneic BALB/cJ mice using an annexin A5 (ANXA5) functionalized SWCNT bioconjugate synergistically enhances an anti-cytotoxic T-lymphocyte-associated protein 4 (anti-CTLA-4)-dependent abscopal response, resulting in an increased survival (55%) at 100 days after tumor inoculation. In comparison, there was no survival at 100 days for either photothermal therapy by itself or immunostimulation by itself. Prior to photothermal therapy, the SWCNT-ANXA5 bioconjugate was administered systemically at a relatively low dose of 1.2 mg/kg, where it then accumulated in tumor vasculature via ANXA5-dependent binding. During photothermal therapy, the average maximum temperature in the tumor reached 54 °C (duration 175 s). The mechanism of prolonged survival resulting from combinatorial photothermal ablation and immune stimulation was evaluated by flow cytometric quantification of splenic antitumoral immune effector cells and serum cytokine quantification.

Pre- and Post-Conditioning of the Heart: An Overview of Cardioprotective Signaling Pathways.

Since the discovery of ischemic pre- and post-conditioning, more than 30 years ago, the knowledge about the mechanisms and signaling pathways involved in these processes has significantly increased. In clinical practice, on the other hand, such advancement has yet to be seen. This article provides an overview of ischemic pre-, post-, remote, and pharmacological conditioning related to the heart. In addition, we reviewed the cardioprotective signaling pathways and therapeutic agents involved in the above-mentioned processes, aiming to provide a comprehensive evaluation of the advancements in the field. The advancements made over the last decades cannot be ignored and with the exponential growth in techniques and applications. The future of pre- and post-conditioning is promising.

Correlações entre maternidade na adolescência e bebê prematuro e de baixo peso: uma revisão integrativa / Correlations between maternity in adolescence and the birth of a premature and low weight baby: an integrative literature review

O presente trabalho apresenta uma revisão integrativa da produção científica nacional e internacional, dos últimos cinco anos, sobre estudos existentes na literatura que abordem a correlação entre a maternidade na adolescência ao nascimento de um bebê prematuro e de baixo peso. Foram acessadas publicações indexadas nas bases de dados Lilacs, Pubmed Central, SciELO (Web of Science), e Scopus (Elsevier), através dos descritores: adolescent, mother, baby ou infant newborn,preterm e low birth weight em artigos de língua inglesa e portuguesa. Entre 206 publicações encontradas, 11 trabalhos foram selecionados de acordo com os critérios de inclusão e por avaliação entre juízes, os quais permitiram compreender temáticas especificas da maternidade adolescente frente ao parto prematuro e de baixo peso ao nascimento. Algumas temáticas foram destacadas como: aspectos emocionais das mães, questões psicossociais, associação entre IMC (índice de massa corporal - peso) materno (pré e peri gestacional) as taxas relativas do nascimento prematuro e ou baixo peso do bebê e o risco da maternidade adolescente e suas comorbidades. Conclui-se que programas de acompanhamento para prevenção da gravidez adolescente, educação em saúde são essenciais para minimizar as taxas de nascimentos prematuros e com baixo peso, assim como a assistência pré-natal direcionada a problemática relativa a condição de saúde do adolescente.

The present work presents an integrative review of the national and international scientific production, of the last five years, on existing studies in the literature that address the correlation between motherhood in adolescence and the birth of a premature and low weight baby. Publications indexed in the Lilacs, Pubmed Central, SciELO (Web of Science), and Scopus (Elsevier) databases were accessed through the descriptors: adolescent, mother, baby or infant newborn, preterm and low birth weight, in English and Portuguese languages. Among 206 publications found, 11 papers were selected according to the inclusion criteria and by evaluation among judges, which allowed to understand specific themes of adolescent motherhood in the face of premature birth and low birth weight. Some themes were highlighted as: emotional aspects of mothers, psychosocial issues, association between maternal BMI (body mass index - weight) (pre and peri gestational), relative rates of premature birth and or low weight of the baby and the risk of motherhood adolescent and his comorbidities. It is concluded that follow-up programs for the prevention of adolescent pregnancy, health education are essential to minimize the rates of premature births and low birth weight, as well as prenatal care directed to problems related to the adolescent's health condition.

Functional polymorphisms of BCL11A and HBS1L-MYB genes affect both fetal hemoglobin level and clinical outcomes in a cohort of children with sickle cell anemia.

Fetal hemoglobin (HbF) ameliorates clinical severity of sickle cell anemia (SCA). The major loci regulating HbF levels are HBB cluster, BCL11A, and HMIP-2 (HBS1L-MYB). However, the impact of noncoding single-nucleotide polymorphisms (SNPs) in these loci on clinical outcomes and their functional role on regulating HbF levels should be better elucidated. Therefore, we performed comprehensive association analyses of 14 noncoding SNPs in five loci with HbF levels and with clinical outcomes in a cohort of 250 children with SCA from Southeastern Brazil, and further performed functional annotation of these SNPs. We found SNPs independently associated with HbF levels: rs4671393 in BCL11A (ß-coefficient = 0.28), rs9399137 in HMIP-2A (ß-coefficient = 0.16), and rs4895441 in HMIP-2B (ß-coefficient = 0.15). Patients carrying minor (HbF-boosting) alleles for rs1427407, rs93979137, rs4895441, rs9402686, and rs9494145 showed reduced count of reticulocytes (p < 0.01), while those carrying the T allele of rs9494145 showed lower white blood cell count (p = 0.002). Carriers of the minor allele for rs9402686 showed higher peripheral saturation of oxygen (p = 0.002). Patients carrying minor alleles in BCL11A showed lower risk of transfusion incidence rate ratio (IRR ≥ 1.3; p < 0.0001). This effect was independent of HbF effect (p = 0.005). Carriers of minor alleles for rs9399137 and rs9402686 showed lower risk of acute chest syndrome (IRR > 1.3; p ≤ 0.01). Carriers of the reference allele for rs4671393 showed lower risk of infections (IRR = 1.16; p = 0.01). In conclusion, patients carrying HbF-boosting alleles of BCL11A and HMIP-2 were associated with milder clinical phenotypes. Higher HbF concentration may underlie this effect.

Percepção das parturientes sobre violência obstétrica: a dor que querem calar / Parturients' perception of obstetric violence: the pain they want to silence / Percepción de las parturientas acerca de la violencia obstétrica: el dolor que quieren silenciar

Objetivou-se identificar o conhecimento das parturientes sobre violência obstétrica, levantar se conseguem identificar as principais ações presentes na violência obstétrica, detectar os impactos físicos e psicológicos da violência obstétrica. Os sujeitos desse estudo foram 14 puérperas residentes do estado do Rio de Janeiro. Foi realizado um estudo do tipo descritivo, exploratório com abordagem quali-quantitativa. A coleta de dados foi realizada por meio de um formulário eletrônico online, na qual foi viabilizado um formulário individual composto de perguntas relacionadas ao objetivo da pesquisa sendo de fácil e rápido acesso ao sujeito participante. Concluiu-se através dos resultados desta pesquisa que as mulheres possuem um conhecimento limitado acerca da violência obstétrica, podendo estar relacionado à falta de informação durante o pré-natal.(AU)

The aim was to identify the parturients' knowledge about obstetric violence, to get up if they can identify the main actions present in obstetric violence, to detect the physical and psychological impacts of obstetric violence. The subjects of this study were 14 puerperal women living in the state of Rio de Janeiro. A descriptive, exploratory study with a qualitative and quantitative approach was carried out. Data collection was performed using an online electronic form, in which an individual form was made available, consisting of questions related to the research objective, being easily and quickly accessed by the participating subject. It was concluded through the results of this research that women have limited knowledge about obstetric violence, which may be related to the lack of information during prenatal care.(AU)

El objetivo era identificar el conocimiento de las parturientas sobre la violencia obstétrica, levantarse si pueden identificar las principales acciones presentes en la violencia obstétrica, detectar los impactos físicos y psicológicos de la violencia obstétrica. Los sujetos de este estudio fueron 14 mujeres puerperales que viven en el estado de Rio de Janeiro. Se realizó un estudio exploratorio descriptivo con enfoque cualitativo y cuantitativo. La recopilación de datos se realizó mediante un formulario electrónico en línea, en el que se puso a disposición un formulario individual, que consta de preguntas relacionadas con el objetivo de la investigación, de fácil y rápido acceso por parte del sujeto participante. A través de los resultados de esta investigación, se concluyó que las mujeres tienen un conocimiento limitado sobre la violencia obstétrica, que puede estar relacionada con la falta de información durante la atención prenatal.(AU)

Injectable, porous, biohybrid hydrogels incorporating decellularized tissue components for soft tissue applications.

Biodegradable injectable hydrogels have been extensively studied and evaluated in various medical applications such as for bulking agents, drug delivery reservoirs, temporary barriers, adhesives, and cell delivery matrices. Where injectable hydrogels are intended to facilitate a healing response, it may be desirable to encourage rapid cellular infiltration into the hydrogel volume from the tissue surrounding the injection site. In this study, we developed a platform technique to rapidly form pores in a thermally responsive injectable hydrogel, poly(NIPAAm-co-VP-co-MAPLA) by using mannitol particles as porogens. In a rat hindlimb muscle injection model, hydrogels incorporating porosity had significantly accelerated cellular infiltration. To influence the inflammatory response to the injected hydrogel, enzymatically digested urinary bladder matrix (UBM) was mixed with the solubilized hydrogel. The presence of UBM was associated with greater polarization of the recruited macrophage population to the M2 phenotype, indicating a more constructive foreign body response. The hybrid hydrogel positively affected the wound healing outcomes of defects in rabbit adipose tissue with negligible inflammation and fibrosis, whereas scar formation and chronic inflammation were observed with autotransplantation and in saline injected groups. These results demonstrate the value of combining the effects of promoting cell infiltration and mediating the foreign body response for improved biomaterials options soft tissue defect filling applications. STATEMENT OF SIGNIFICANCE: Our objective was to develop a fabrication process to create porous injectable hydrogels incorporating decellularized tissue digest material. This new hydrogel material was expected to exhibit faster cellular infiltration and a greater extent of pro-M2 macrophage polarization compared to control groups not incorporating each of the functional components. Poly(NIPAAm-co-VP-co-MAPLA) was chosen as the representative thermoresponsive hydrogel, and mannitol particles and digested urinary bladder matrix (UBM) were selected as the porogen and the bioactive decellularized material components respectively. In rat hindlimb intramuscular injection models, this new hydrogel material induced more rapid cellular infiltration and a greater extent of M2 macrophage polarization compared to control groups not incorporating all of the functional components. The hybrid hydrogel positively affected the wound healing outcomes of defects in rabbit adipose tissue with negligible inflammation and fibrosis, whereas scar formation and chronic inflammation were observed with autotransplantation and in saline injected groups. The methodology of this report provides a straightforward and convenient mechanism to promote cell infiltration and mediate foreign body response in injectable hydrogels for soft tissue applications. We believe that the readership of Acta Biomaterialia will find the work of interest both for its specific results and general translatability of the findings.

Cancer Progenitor Cells: The Result of an Epigenetic Event?

The concept of cancer stem cells was proposed in the late 1990s. Although initially the idea seemed controversial, the existence of cancer stem cells is now well established. However, the process leading to the formation of cancer stem cells is still not clear and thus requires further research. This article discusses epigenetic events that possibly produce cancer progenitor cells from predisposed cells by the influence of their environment. Every somatic cell possesses an epigenetic signature in terms of histone modifications and DNA methylation, which are obtained during lineage-specific differentiation of pluripotent stem cells, which is specific to that particular tissue. We call this signature an epigenetic switch. The epigenetic switch is not fixed. Our epigenome alters with aging. However, depending on the predisposition of the cells of a particular tissue and their microenvironment, the balance of the switch (histone modifications and the DNA methylation) may be tilted to immortality in a few cells, which generates cancer progenitor cells.

Fontes de autoeficácia docente de universitários de educação física / Sources of self-efficacy of pre-service teachers in physical education

RESUMO Este estudo analisa as fontes de autoeficácia docente de universitários em Educação Física. Foram empregados procedimentos de pesquisa qualitativa de caráter descritivo-diagnóstico. Os dados foram obtidos através de entrevista semiestruturada, baseada nos instrumentos: escala de autoeficácia docente e escala de fontes de autoeficácia docente, com 7 universitários da última fase do curso de licenciatura em Educação Física de uma universidade pública no Estado de Santa Catarina. Para a análise dos casos múltiplos, utilizou-se a técnica de análise de conteúdo, que consiste, primeiramente, na transcrição e descrição de cada caso individualmente, seguida do cruzamento e confronto das informações dos casos. Os resultados mostraram que as fontes de autoeficácia, referentes ao manejo de classe, foram: experiência de domínio (6), persuasão social (5) e vicária (4). Sobre a intencionalidade docente, as fontes mais citadas foram: experiências vicárias (7), persuasão social (7) e domínio (6). Especificamente sobre a intencionalidade docente, os universitários mencionaram uma maior diversidade de situações de autoeficácia frente ao manejo de classe. Conclui-se que a observação e a persuasão social foram as fontes de autoeficácia consensualmente valorizadas pelos universitários. Essas situações foram proporcionadas pela universidade nos estágios obrigatórios, observando e recebendo orientações dos professores da universidade e dos professores da escola.

ABSTRACT The aim of this study was to analyze the sources of self-efficacy of pre-service teachers in physical education. Were employed qualitative research procedures and the data were collected through semistructured interview with 7 pre-service teachers in Physical Education from a public university. For the analysis of multiple cases was used the content analysis technique, which consists, first, in transcription and description of each case individually, then crossing and confrontation of information for all cases. The results showed that the sources of self-efficacy for the class management were the mastery experiences (6), social persuasion (5) and vicarious (4). About the teaching intentionality, the most cited sources were the vicarious experiences (7), social persuasion (7) and the mastery experiences (6). Specifically on teaching intentionality, the pre-service teachers mentioned more variety of situations of self-efficacy when compared to the management class. The observation and social persuasion were the sources of self-efficacy commonly valued by students. These situations were provided in the required stages, watching and getting supervision from teachers at the University and teachers from school.

Congenital Toxoplasmosis in Chronically Infected and Subsequently Challenged Ewes.

This experiment studied congenital transmission in sheep experimentally infected with oocysts of Toxoplasma gondii and reinfected at one of three stages of pregnancy. Twenty ewes were experimentally infected with T. gondii strain ME49 (day 0). After the T. gondii infection became chronic (IFAT≤512), the ewes were allocated with rams for coverage. After the diagnosis of pregnancy, these ewes were allocated into four experimental groups (n = 5): I-reinfected with T. gondii on the 40th day of gestation (DG); II-reinfected on DG 80; III-reinfected on DG 120; and IV-saline solution on DG 120 (not reinfected). Five ewes (IFAT<64) were kept as negative controls (uninfected, group V), therefore in groups I-III were infected prior to pregnancy and re-infected during pregnancy, group IV was only infected prior to pregnancy, and group V was not infected. Parasitism by T. gondii was investigated (histopathology, immunohistochemistry, mouse bioassay and PCR) in mothers and lambs tissue. All ewes produced lambs serologically positive for T. gondii. The results of the mouse bioassay, immunohistochemistry and PCR assays revealed the presence of T. gondii in all 20 sheep and their lambs. The congenital transmission of T. gondii was associated with fetal loss and abnormalities in persistently infected sheep and in ewes infected and subsequently reinfected by this protozoan. Therefore, congenital T. gondii infection was common when ewes were chronically infected prior to pregnancy, with or without reinfection during at various stages of gestation.

Exposure of cultured fibroblasts to the peptide PR-11 for the identification of induced proteome alterations and discovery of novel potential ligands.

The PR-11 peptide corresponds to the N-terminal and active region of the endogenously synthesized PR-39 molecule, of porcine origin. It is known to possess various biological effects including antimicrobial properties, angiogenic and anti-inflammatory activities. Apart from its reported activity as a proteasome inhibitor, a more comprehensive understanding of its function, at the molecular level, is still lacking. In this study, we used a label-free shotgun strategy to evaluate the proteomic alterations caused by exposure of cultured fibroblasts to the peptide PR-11. This approach revealed that more than half of the identified molecules were related to signalling, transcription and translation. Proteins directly associated to regulation of angiogenesis and interaction with the hypoxia-inducible factor 1-α (HIF-1α) were significantly altered. In addition, at least three differentially expressed molecules of the NF-κB pathway were detected, suggesting an anti-inflammatory property of PR-11. At last, we demonstrated novel potential ligands of PR-11, through its immobilization for affinity chromatography. Among the eluted molecules, gC1qR, a known complement receptor, appeared markedly enriched. This provided preliminary evidence of a PR-11 ligand possibly involved in the internalization of this peptide. Altogether, our findings contributed to a better understanding of the cellular pathways affected by PR-39 derived molecules.