Safety and Performance of a Cell-Impermeable Endoprosthesis for Hemodialysis Vascular Access Outflow Stenosis: A Brazilian Multicenter Retrospective Study.

PURPOSE: To evaluate the safety and performance of Wrapsody™, a cell-impermeable endoprosthesis (CIE), for treating hemodialysis vascular access outflow stenosis. MATERIALS AND METHODS: Investigators retrospectively analyzed 113 hemodialysis patients treated with a CIE (11/2021-12/2022) across four centers in Brazil. De novo or restenotic lesions were treated. The primary efficacy outcome measure was target lesion primary patency (TLPP) at 1, 3, 6, and 12 months; the primary safety outcome measure was the absence of serious local or systemic adverse events within the first 30 days post-procedure. Secondary outcome measures included technical and procedural success, access circuit primary patency (ACPP), and secondary patency at 1, 3, 6, and 12 months post-procedure. RESULTS: Thirty-nine patients (34.5%) had thrombosed access at the initial presentation, and 38 patients (33.6%) presented with recurrent stenosis. TLPP rates at 1, 3, 6, and 12 months were 100%, 96.4%, 86.4%, and 69.7%, respectively. ACPP rates were 100% at 1 month, 89.2% at 3 months, 70.9% at 6 months, and 56.0% at 12 months. The target lesion secondary patency rates at 1, 3, 6, and 12 months were 100%, 97.3%, 93.6%, and 91.7%, respectively. In the adjusted multivariate Cox regression analysis, male sex and endoprosthesis with diameters of 10, 12, 14, and 16 mm were associated with improved primary patency rates. No localized or systemic serious adverse event was observed through the first 30 days post-procedure. CONCLUSION: The CIE evaluated in this study is safe and effective for treating peripheral and central outflow stenoses in hemodialysis vascular access. LEVEL OF EVIDENCE: Level 2b, cohort study.

IL-1R signaling drives enteric glia-macrophage interactions in colorectal cancer.

Enteric glia have been recently recognized as key components of the colonic tumor microenvironment indicating their potential role in colorectal cancer pathogenesis. Although enteric glia modulate immune responses in other intestinal diseases, their interaction with the colorectal cancer immune cell compartment remains unclear. Through a combination of single-cell and bulk RNA-sequencing, both in murine models and patients, here we find that enteric glia acquire an immunomodulatory phenotype by bi-directional communication with tumor-infiltrating monocytes. The latter direct a reactive enteric glial cell phenotypic and functional switch via glial IL-1R signaling. In turn, tumor glia promote monocyte differentiation towards pro-tumorigenic SPP1+ tumor-associated macrophages by IL-6 release. Enteric glia cell abundancy correlates with worse disease outcomes in preclinical models and colorectal cancer patients. Thereby, our study reveals a neuroimmune interaction between enteric glia and tumor-associated macrophages in the colorectal tumor microenvironment, providing insights into colorectal cancer pathogenesis.

Safety and efficacy of a new covered stent in hemodialysis vascular access outflow stenosis: A Brazilian multicenter retrospective study.

BACKGROUND: Vascular stenosis commonly leads to dysfunction in hemodialysis vascular access. Although percutaneous transluminal angioplasty is an established treatment, stent utilization has increased in the last decade as an alternative solution to extend the access function. This study evaluated the safety and initial results of a new impermeable covered stent for treating vascular access outflow stenosis. METHODS: Investigators retrospectively analyzed 114 hemodialysis patients treated with polytetrafluorethylene-covered stents from September 2018 to September 2022 across four centers. Lesions treated were de novo or restenotic and located in the venous graft anastomosis, outflow segment, cephalic arch, and basilic swing point. Patients were followed by in-person physical examination at 1, 3, and 6 months, and Duplex ultrasound was performed to evaluate the vascular access circuit and in-stent restenosis. The primary efficacy endpoint was target lesion primary patency at 1, 3, and 6 months. Secondary endpoints included access circuit primary patency and secondary patency at 1, 3, and 6 months. The primary safety endpoint was freedom from local or systemic serious adverse events through 30 days post-procedure. RESULTS: Forty-four patients had thrombosed access at the initial presentation, and 41 patients presented with recurrent stenosis. The target lesion primary patency rates at 1, 3, and 6 months were 100%, 89.4%, and 74%, respectively. The access circuit primary patency rates were 100% at 1 month, 85% at 3 months, and 62.7% at 6 months. The secondary patency rates at 1, 3, and 6 months were 100%, 96.4%, and 94.6%, respectively. In the adjusted multivariate Cox regression analysis, only recurrent lesions and female gender were associated with reduced primary patency rates. No serious adverse event was observed through the first 30 days post-procedure. CONCLUSION: In this retrospective analysis, a new covered stent was shown to be safe and effective for treating peripheral outflow stenosis in vascular access.

Sf9 Cell Metabolism Throughout the Recombinant Baculovirus and Rabies Virus-Like Particles Production in Two Culture Systems.

This work aimed to assess the Sf9 cell metabolism during growth, and infection steps with recombinant baculovirus bearing rabies virus proteins, to finally obtain rabies VLP in two culture systems: Schott flask (SF) and stirred tank reactor (STR). Eight assays were performed in SF and STR (four assays in each system) using serum-free SF900 III culture medium. Two non-infection growth kinetics assays and six recombinant baculovirus infection assays. The infection runs were carried out at 0.1 pfu/cell multiplicity of infection (MOI) for single baculovirus bearing rabies glycoprotein (BVG) and matrix protein (BVM) and a coinfection with both baculoviruses at MOI of 3 and 2 pfu/cell for BVG and BVM, respectively. The SF assays were done in triplicate. The glucose, glutamine, glutamate, lactate, and ammonium uptake or release specific rates were quantified over the exponential growth phase and infection stage. The highest uptake specific rate was observed for glucose (42.5 × 10-12 mmol cell/h) in SF and for glutamine (30.8 × 10-12 mmol/cell/h) in STR, in the exponential growth phases. A wave pattern was observed for assessed analytes throughout the infection phase and the glucose had the highest wave amplitude within the 10-10 mmol cell/h order. This alternative uptake and release behavior is in harmony with the lytic cycle of baculovirus in insect cells. The virus propagation and VLP generation were not limited by glucose, glutamine, and glutamate, neither by the toxicity of lactate nor ammonium under the conditions appraised in this work. The findings from this work can be useful to set baculovirus infection processes at high cell density to improve rabies VLP yield, purity, and productivity.

Zika virus-like particles (VLPs) produced in insect cells.

Introdutcion: The Zika virus (ZIKV) infections are a healthcare concern mostly in the Americas, Africa, and Asia but have increased its endemicity area beyond these geographical regions. Due to the advances in infections by Zika virus, it is imperative to develop diagnostic and preventive tools against this viral agent. Virus-like particles (VLPs) appear as a suitable approach for use as antiviral vaccines. Methods: In this work, a methodology was established to produce virus-like particles containing the structural proteins, C, prM, and E of Zika virus produced in insect cells using the gene expression system derived from baculovirus. The vector pFast- CprME -ZIKV was constructed containing the gene sequences of Zika virus structural proteins and it was used to generate the recombinant bacmids (Bac- CprME -ZIKV) through transformation into DH10BacTM cells. The Bac- CprME -ZIKV was transfected in Spodoptera frugiperda (Sf9) insect cells and batches of BV- CprME -ZIKV were obtained by infection assays using a multiplicity of infection of 2. The Sf9 cells were infected, and the supernatant was collected 96 h post-infection. The expression of the CprME -ZIKV protein on the cell surface could be observed by immunochemical assays. To concentrate and purify virus-like particles, the sucrose and iodixanol gradients were evaluated, and the correct CprME -ZIKV proteins' conformation was evaluated by the Western blot assay. The virus-like particles were also analyzed and characterized by transmission electron microscopy. Results and discussion: Spherical structures like the native Zika virus from 50 to 65 nm containing the CprME -ZIKV proteins on their surface were observed in micrographs. The results obtained can be useful in the development path for a vaccine candidate against Zika virus.

Exposure to Mancozeb results in increased MAPK phosphorylation and locomotor deficits in zebrafish larvae.

Mancozeb is a widely used fungicide whose toxicity has been reported in non-target organisms, being considered to have high or very high acute toxicity to aquatic organisms. However, the toxicity of this compound is not well characterized in the developmental stages of fish. In this study, Danio rerio with 4-, 5-, and 6-days post fertilization (dpf) was exposed to MZ at non-lethal concentrations for 24, 48, or 72 h and subsequently, behavioral alterations, oxidative stress parameters and ERK, p38MAPK, and Akt phosphorylation were analyzed. MZ exposure during the larval period decreased motor performance evaluated by traveled distance, immobile time, and time spent in the peripheral area. In parallel, MZ induced ROS levels and increased the number of cells in apoptosis, causing severe DNA damage, inducing Acetylcholinesterase and Superoxide dismutase activities, and inhibiting Glutathione peroxidase and thioredoxin reductase. Additionally, phosphorylation levels of the proteins p38MAPK, ERK2, and Akt were stimulated. These findings are relevant considering the ecological implications of MZ exposure to fishes in different developmental stages and the role of the MAPK pathway in events like development and cell death.

Biochemical monitoring throughout all stages of rabies virus-like particles production by Raman spectroscopy using global models.

This work aimed to quantify growth and biochemical parameters (viable cell density, Xv; cell viability, CV; glucose, lactate, glutamine, glutamate, ammonium, and potassium concentrations) in upstream stages to obtain rabies virus-like particles (rabies VLP) from insect cell-baculovirus system using on-line and off-line Raman spectra to calibrate global models with minimal experimental data. Five cultivations in bioreactor were performed. The first one comprised the growth of uninfected Spodoptera frugiperda (Sf9) cells, the second and third runs to obtain recombinant baculovirus (rBV) bearing Rabies G glycoprotein and matrix protein, respectively. The fourth one involved the generation of rabies VLP from rBVs and the last one was a repetition of the third one with cell inoculum infected by rBV. The spectra were acquired through a Raman spectrometer with a 785-nm laser source. The fitted Partial Least Square models for nutrients and metabolites were comparable with those previously reported for mammalian cell lines (Relative error < 15 %). However, the use of this chemometrics approach for Xv and CV was not as accurate as it was for other parameters. The findings from this work established the basis for bioprocess Raman spectroscopical monitoring using insect cells for VLP manufacturing, which are gaining ground in the pharmaceutical industry.

Oversized balloon angioplasty for endovascular maturation of arteriovenous fistulae to accelerate cannulation and to decrease the duration of catheter use.

BACKGROUND: The functional autologous arteriovenous fistula is considered the best vascular access for patients on hemodialysis. Some fistulae exhibit maturation problems after creation and do not reach adequate vessel diameter and flow in dialysis. The aim of this study was to describe our technique of oversized balloon angioplasty for assisted maturation of arteriovenous fistulae to accelerate the cannulation and to decrease the time of catheter use and its outcome. METHODS: A retrospective analysis of balloon-assisted procedures for maturation failure was performed in a single center between October 2011 and January 2019. Patients underwent imaging procedures to identify stenosis, followed by angioplasty using oversized high-pressure balloons from the anastomosis to the deep venous outflow tract. The flow volume, time interval of use of the fistula and removal of the catheter, patency rates, and complications rates were evaluated. RESULTS: Seventy-eight patients underwent 124 balloon angioplasty procedures. Technical and clinical success was achieved in 91% of the cases. In patients in whom maturation was successful, the fistula was cannulated in a mean time of 5 days after the procedure (range, 1-20 days). On average, catheter removal was performed 14 days (range, 5-33 days) after the maturation procedure. The mean flow volume in the fistula before the procedure was 276 ml/min (range, 122-488 ml/min) and 24 h after the maturation was 1014 ml/min (range, 760-1800 ml/min).The primary patency rate at 3, 6, and 12 months was 87.3%, 66.2%, and 50.7%, respectively. Assisted primary patency was 100% at 3 months, 92.9% at 6 months, and 90.0% at 12 months. Minor complications occurred in 18% of cases, and major complications in 4.8%. CONCLUSIONS: The oversized balloon-assisted arteriovenous fistula maturation technique is safe and effective, allowing the cannulation of the fistulae a few hours or days after the procedure and decreasing the time of catheter use.

Analgesic and anti-inflammatory activities of ethyl acetate and butanol fractions from Vitex polygama hydroalcoholic leaf extract.

This study evaluated the analgesic and anti-inflammatory activities of Vitex polygama. Ethyl acetate and butanol fractions (10-30 mg/kg), obtained from the hydroalcoholic leaf extract, showed an antinociceptive effect in the acetic acid-induced abdominal writhing test, formalin test and modified hot plate test in mice, indicating a peripheral anti-inflammatory action. Ethyl acetate and butanol fractions were effective in inhibiting nitric oxide and TNF-α production, respectively, in RAW 264.7 macrophages. Both fractions (10-30 mg/kg) showed an acute analgesic effect in mice with vincristine-induced neuropathic pain exposed to a thermal stimulus. Through ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-UV-MS/MS) it was possible to identify seven major compounds: isoorientin, orientin, vitexin, isovitexin, O-p-hydroxybenzoyl orientin, O-caffeoyl-orientin, and di-caffeoylquinic acid. Orientin and isoorientin were isolated from ethyl acetate fraction and had their identity confirmed by nuclear magnetic resonance (NMR). Glucosyl flavones appear to be the main metabolites responsible for the anti-inflammatory and analgesic activities observed for V. polygama.

Participação paterna no pré-natal, parto e pós-parto: um estudo sobre a perspectiva do pai / Paternal participation in antenatal, labour and postpartum: a study about the father's perspective / Participación paterna en pré-natal, parto y pos-natal: un estudio sobre la perspectiva del padre

Este estudo parte de uma exploração mais ampla sobre as transformações do exercício paterno na contemporaneidade. O objetivo foi investigar a participação do pai nas consultas pré-natais, no parto e no pós-parto sob a perspectiva masculina. Para tanto, realizou-se uma pesquisa qualitativa, na qual foram entrevistados quinze pais primíparos. Os resultados foram analisados utilizando o método de análise de conteúdo na sua vertente categorial. Da análise do material, emergiram diversas categorias de análise. Contudo, para atingir os objetivos formulados no presente trabalho, foram discutidas as categorias "presença do pai nas consultas de pré-natal", "presença do pai na sala de parto" e "rede de apoio familiar". Os resultados indicam que, na atualidade, muitos homens desejam participar do ciclo gravídico-puerperal. Entretanto, sua presença nem sempre é facilitada, em especial, devido aos estereótipos de gênero associados ao universo obstetrício como um espaço naturalmente feminino. A partir da análise das entrevistas concluiu-se que a participação do homem durante o pré-natal, parto e pós-parto é um fator positivo, pois fortalece os vínculos familiares

This study, part of a broader investigation on the transformations of fatherly exercise in contemporary times, and aimed to investigate the father's participation in prenatal consultations, childbirth and postpartum from the man's perspective. Therefore, a qualitative research was conducted, with fifteen primiparous parents wich were interviewed. The results were analyzed according to the content analysis method in each categorical aspect. From the analysis of the material, several categories of analysis emerged. To achieve the objectives formulated in this study, the categories presence of the father in prenatal consultations, presence of the father in the delivery room and family support network will be discussed. The results showed that, today, many men wish to participate in the pregnancy-puerperal cycle, however their presence is not always facilitated, especially due to the gender stereotypes associated with the obstetrical universe as a naturally female space. This concludes that the participation of men during prenatal, delivery and postpartum is a positive factor, because strengthens family bonds

Este estudio, parte de una investigación más amplia sobre las transformaciones del ejercicio paterno en la época contemporánea, tuvo como objetivo investigar la participación del padre en las consultas prenatales, el parto y el pós-parto desde la perspectiva del hombre. Para ello se realizó una investigación cualitativa, en la que se entrevistó a quince padres primíparos. Los resultados se analizaron según el método de análisis de contenido en su aspecto categórico. Del análisis del material surgieron varias categorías de análisis. Para lograr los objetivos formulados en este estudio se discutirán las categorías presencia del padre en consultas prenatales, presencia del padre en la sala de partos y red de apoyo familiar. Los resultados mostraron que, hoy en día, muchos hombres desean participar en el ciclo embarazopuerperal, sin embargo su presencia no siempre se facilita, especialmente por los estereotipos de género asociados al universo obstétrico como un espacio naturalmente femenino. Se concluye que la participación de los hombres durante el prenatal, parto y posparto es un factor positivo, ya que fortalece los lazos familiares

Zika virus-like particles (VLPs) produced in insect cells

Introdutcion: The Zika virus (ZIKV) infections are a healthcare concern mostly in the Americas, Africa, and Asia but have increased its endemicity area beyond these geographical regions. Due to the advances in infections by Zika virus, it is imperative to develop diagnostic and preventive tools against this viral agent. Virus-like particles (VLPs) appear as a suitable approach for use as antiviral vaccines. Methods: In this work, a methodology was established to produce virus-like particles containing the structural proteins, C, prM, and E of Zika virus produced in insect cells using the gene expression system derived from baculovirus. The vector pFast- CprME -ZIKV was constructed containing the gene sequences of Zika virus structural proteins and it was used to generate the recombinant bacmids (Bac- CprME -ZIKV) through transformation into DH10BacTM cells. The Bac- CprME -ZIKV was transfected in Spodoptera frugiperda (Sf9) insect cells and batches of BV- CprME -ZIKV were obtained by infection assays using a multiplicity of infection of 2. The Sf9 cells were infected, and the supernatant was collected 96 h post-infection. The expression of the CprME -ZIKV protein on the cell surface could be observed by immunochemical assays. To concentrate and purify virus-like particles, the sucrose and iodixanol gradients were evaluated, and the correct CprME -ZIKV proteins’ conformation was evaluated by the Western blot assay. The virus-like particles were also analyzed and characterized by transmission electron microscopy. Results and discussion: Spherical structures like the native Zika virus from 50 to 65 nm containing the CprME -ZIKV proteins on their surface were observed in micrographs. The results obtained can be useful in the development path for a vaccine candidate against Zika virus.

Sf9 cell metabolism throughout the recombinant Baculovirus and rabies virus-like particles production in two culture systems

This work aimed to assess the Sf9 cell metabolism during growth, and infection steps with recombinant baculovirus bearing rabies virus proteins, to fnally obtain rabies VLP in two culture systems: Schott fask (SF) and stirred tank reactor (STR). Eight assays were performed in SF and STR (four assays in each system) using serum-free SF900 III culture medium. Two non-infection growth kinetics assays and six recombinant baculovirus infection assays. The infection runs were carried out at 0.1 pfu/cell multiplicity of infection (MOI) for single baculovirus bearing rabies glycoprotein (BVG) and matrix protein (BVM) and a coinfection with both baculoviruses at MOI of 3 and 2 pfu/cell for BVG and BVM, respectively. The SF assays were done in triplicate. The glucose, glutamine, glutamate, lactate, and ammonium uptake or release specifc rates were quantifed over the exponential growth phase and infection stage. The highest uptake specifc rate was observed for glucose (42.5× 10–12 mmol cell/h) in SF and for glutamine (30.8× 10–12 mmol/cell/h) in STR, in the exponential growth phases. A wave pattern was observed for assessed analytes throughout the infection phase and the glucose had the highest wave amplitude within the 10–10 mmol cell/h order. This alternative uptake and release behavior is in harmony with the lytic cycle of baculovirus in insect cells. The virus propagation and VLP generation were not limited by glucose, glutamine, and glutamate, neither by the toxicity of lactate nor ammonium under the conditions appraised in this work. The fndings from this work can be useful to set baculovirus infection processes at high cell density to improve rabies VLP yield, purity, and productivity.

Biochemical monitoring throughout all stages of rabies virus-like particles production by Raman spectroscopy using global models

This work aimed to quantify growth and biochemical parameters (viable cell density, Xv; cell viability, CV; glucose, lactate, glutamine, glutamate, ammonium, and potassium concentrations) in upstream stages to obtain rabies virus-like particles (rabies VLP) from insect cell-baculovirus system using on-line and off-line Raman spectra to calibrate global models with minimal experimental data. Five cultivations in bioreactor were performed. The first one comprised the growth of uninfected Spodoptera frugiperda (Sf9) cells, the second and third runs to obtain recombinant baculovirus (rBV) bearing Rabies G glycoprotein and matrix protein, respectively. The fourth one involved the generation of rabies VLP from rBVs and the last one was a repetition of the third one with cell inoculum infected by rBV. The spectra were acquired through a Raman spectrometer with a 785-nm laser source. The fitted Partial Least Square models for nutrients and metabolites were comparable with those previously reported for mammalian cell lines (Relative error < 15 %). However, the use of this chemometrics approach for Xv and CV was not as accurate as it was for other parameters. The findings from this work established the basis for bioprocess Raman spectroscopical monitoring using insect cells for VLP manufacturing, which are gaining ground in the pharmaceutical industry.

Performance comparison of recombinant Baculovirus and rabies virus-like particles production using two culture platforms

This work aimed to assess, following upstream optimization in Schott flasks, the scalability from this culture platform to a stirred-tank bioreactor in order to yield rabies-recombinant baculovirus, bearing genes of G (BVG) and M (BVM) proteins, and to obtain rabies virus-like particles (VLP) from them, using Sf9 insect cells as a host. Equivalent assays in Schott flasks and a bioreactor were performed to compare both systems and a multivariate statistical approach was also carried out to maximize VLP production as a function of BVG and BVM’s multiplicity of infection (MOI) and harvest time (HT). Viable cell density, cell viability, virus titer, BVG and BVM quantification by dot-blot, and BVG quantification by Enzyme-Linked Immunosorbent Assay (ELISA) were monitored throughout the assays. Furthermore, transmission electron microscopy was used to characterize rabies VLP. The optimal combination for maximum VLP expression was BVG and BVM MOI of 2.3 pfu/cell and 5.1 pfu/cell, respectively, and 108 h of harvest time. The current study confirmed that the utilization of Schott flasks and a benchtop bioreactor under the conditions applied herein are equivalent regarding the cell death kinetics corresponding to the recombinant baculovirus infection process in Sf9 cells. According to the results, the hydrodynamic and chemical differences in both systems seem to greatly affect the virus and VLP integrity after release.

Oxygen uptake and transfer rates throughout production of recombinant baculovirus and rabies virus-like particles

The technologies used in rabies vaccines manufacturing for human use are based on the inactivated virus platform. An alternative to traditional vaccines is virus-like particles (VLPs). This work aimed to characterize the oxygen uptake and transfer rate parameters throughout recombinant baculovirus (rBV) and rabies VLPs production using Sf9 cells in stirred tank bioreactor (STB) for a better bioprocess understanding and scalability. Four runs in a bench STB were performed: cell culture without infection; cells infected singly with rBV bearing rabies virus glycoprotein (rBVG, multiplicity of infection, MOI=0.1 pfu/cell) and matrix protein (rBVM, MOI=0.1 pfu/cell), and coinfected with BVG and BVM at MOI of 3 and 2 pfu/cell, respectively. The specific oxygen uptake rate () and volumetric oxygen transfer coefficient () were monitored throughout the reactions, as well as viable cell concentration, viability, rBV titers, and protein concentration. According to the results herein, the aeration and agitation systems in a bioreactor at a higher scale could be designed using the criterium for scale-up of constant , without oxygen facilities. Besides, rabies VLPs volumetric yield of 2.8 mg/L with a typical size (55–68 nm) was obtained. These findings suggest a promising bioprocess for rabies VLPs at a commercial scale.

Increment of immunogenicity after third dose of a homologous inactivated SARS-CoV-2 vaccine in a large population of patients with autoimmune rheumatic diseases.

OBJECTIVE: To determine the immunogenicity of the third dose of CoronaVac vaccine in a large population of patients with autoimmune rheumatic diseases (ARD) and the factors associated with impaired response. METHODS: Adult patients with ARD and age-balanced/sex-balanced controls (control group, CG) previously vaccinated with two doses of CoronaVac received the third dose at D210 (6 months after the second dose). The presence of anti-SARS-CoV-2 S1/S2 IgG and neutralising antibodies (NAb) was evaluated previously to vaccination (D210) and 30 days later (D240). Patients with controlled disease suspended mycophenolate mofetil (MMF) for 7 days or methotrexate (MTX) for 2 weekly doses after vaccination. RESULTS: ARD (n=597) and CG (n=199) had comparable age (p=0.943). Anti-S1/S2 IgG seropositivity rates significantly increased from D210 (60%) to D240 (93%) (p<0.0001) in patients with ARD. NAb positivity also increased: 38% (D210) vs 81.4% (D240) (p<0.0001). The same pattern was observed for CG, with significantly higher frequencies for both parameters at D240 (p<0.05). Multivariate logistic regression analyses in the ARD group revealed that older age (OR=0.98, 95% CI 0.96 to 1.0, p=0.024), vasculitis diagnosis (OR=0.24, 95% CI 0.11 to 0.53, p<0.001), prednisone ≥5 mg/day (OR=0.46, 95% CI 0.27 to 0.77, p=0.003), MMF (OR=0.30, 95% CI 0.15 to 0.61, p<0.001) and biologics (OR=0.27, 95% CI 0.16 to 0.46, p<0.001) were associated with reduced anti-S1/S2 IgG positivity. Similar analyses demonstrated that prednisone ≥5 mg/day (OR=0.63, 95% CI 0.44 to 0.90, p=0.011), abatacept (OR=0.39, 95% CI 0.20 to 0.74, p=0.004), belimumab (OR=0.29, 95% CI 0.13 to 0.67, p=0.004) and rituximab (OR=0.11, 95% CI 0.04 to 0.30, p<0.001) were negatively associated with NAb positivity. Further evaluation of COVID-19 seronegative ARD at D210 demonstrated prominent increases in positivity rates at D240 for anti-S1/S2 IgG (80.5%) and NAb (59.1%) (p<0.0001). CONCLUSIONS: We provide novel data on a robust response to the third dose of CoronaVac in patients with ARD, even in those with prevaccination COVID-19 seronegative status. Drugs implicated in reducing immunogenicity after the regular two-dose regimen were associated with non-responsiveness after the third dose, except for MTX. Trial registration number NCT04754698.

Performance Comparison of Recombinant Baculovirus and Rabies Virus-like Particles production Using Two Culture Platforms.

This work aimed to assess, following upstream optimization in Schott flasks, the scalability from this culture platform to a stirred-tank bioreactor in order to yield rabies-recombinant baculovirus, bearing genes of G (BVG) and M (BVM) proteins, and to obtain rabies virus-like particles (VLP) from them, using Sf9 insect cells as a host. Equivalent assays in Schott flasks and a bioreactor were performed to compare both systems and a multivariate statistical approach was also carried out to maximize VLP production as a function of BVG and BVM's multiplicity of infection (MOI) and harvest time (HT). Viable cell density, cell viability, virus titer, BVG and BVM quantification by dot-blot, and BVG quantification by Enzyme-Linked Immunosorbent Assay (ELISA) were monitored throughout the assays. Furthermore, transmission electron microscopy was used to characterize rabies VLP. The optimal combination for maximum VLP expression was BVG and BVM MOI of 2.3 pfu/cell and 5.1 pfu/cell, respectively, and 108 h of harvest time. The current study confirmed that the utilization of Schott flasks and a benchtop bioreactor under the conditions applied herein are equivalent regarding the cell death kinetics corresponding to the recombinant baculovirus infection process in Sf9 cells. According to the results, the hydrodynamic and chemical differences in both systems seem to greatly affect the virus and VLP integrity after release.

Care among siblings and repercussions of generational transmission on the sibling group / Cuidado entre irmãos e repercussões da transmissão geracional na fratria / Cuidado entre hermanos y repercusiones de la transmisión generacional en la fratría

This research is part of a broader study concerning siblings and the family network. Its objective was to investigate care among siblings and the repercussions of generational transmission on the sibling group. It employed clinical-qualitative methodology, centered on preliminary clinical interviews with families during the family assessment period. Clinical data were gathered at the Applied Psychology Service of a private university in Rio de Janeiro. The 14 families who took part in the study had the following configurations: four married-couple families, four stepfamilies, three single-parent families and three separated-parent families. We observed that both reduced parental investment and excessive family-care workload made it difficult to establish a more health-promoting parent-child relationship, often leading to the parentification of one or more of the children, with the assignment of parental functions to them. Flaws in the psychic assimilation of elements of generational transmission also influence in the creation of sibling bonds, inhibiting the development of family ties of solidarity and the establishment of healthy care practices among siblings.

Este estudo faz parte de uma pesquisa mais ampla sobre fratrias e rede familiar. Teve-se como objetivo investigar o cuidado entre irmãos e repercussões da transmissão geracional na fratria. Utilizou-se metodologia clínico-qualitativa, centrada em entrevistas clínicas preliminares com famílias no período de avaliação familiar. Os dados clínicos foram coletados no Serviço de Psicologia Aplicada de uma universidade privada carioca. Participaram da pesquisa 14 famílias, com as seguintes configurações: quatro casadas, quatro recasadas, três monoparentais e três separadas. Constatou-se que tanto o reduzido investimento parental quanto a sobrecarga nos cuidados familiares dificultam o estabelecimento de uma relação parento-filial mais promotora de saúde, muitas vezes levando à parentalização de um ou mais filhos, com atribuição de funções parentais a eles. Falhas na apropriação psíquica de elementos da transmissão geracional também repercutem na constituição do vínculo fraterno, dificultando a construção de laços familiares solidários e o estabelecimento de práticas salutares de cuidado entre os irmãos.

Este estudio es parte de una investigación más amplia sobre fratrías y redes familiares. El objetivo fue investigar el cuidado entre hermanos y las repercusiones de la transmisión generacional en la fratría. Se utilizó metodología clínico-cualitativa, centrada en entrevistas clínicas preliminares a las familias durante el período de evaluación familiar. Los datos clínicos se recolectaron en el Servicio de Psicología Aplicada de una universidad privada en Rio de Janeiro. Catorce familias participaron en la investigación, con las siguientes configuraciones: cuatro casadas, cuatro vueltas a casar, tres monoparentales y tres separados. Se encontró que tanto la menor inversión de los padres como la sobrecarga por los cuidados con la familia dificultan el establecimiento de una relación entre padres e hijos que promueva la salud, lo que a menudo conduce a la crianza de uno o más hijos con atribución de las funciones parentales a ellos. Las fallas en la apropiación psíquica de elementos de transmisión generacional también inciden en la constitución de un vínculo fraterno, dificultando la construcción de lazos familiares solidarios y el establecimiento de prácticas de cuidado saludable entre los hermanos.

Behavioral changes occur earlier than redox alterations in developing zebrafish exposed to Mancozeb.

As agriculture expands to provide food and wellbeing to the world's growing population, there is a simultaneous increasing concern about the use of agrochemicals, which can harm non-target organisms, mainly in the aquatic environment. The fungicide Mancozeb (MZ) has been used on a large-scale and is a potent inducer of oxidative stress. Therefore, there is an urgent need for the development of more sensitive biomarkers designed to earlier biomonitoring of this compound. Here we tested the hypothesis that behavioral changes induced by sublethal MZ concentrations would occur first as compared to biochemical oxidative stress markers. Embryos at 4 h post-fertilization (hpf) were exposed to Mancozeb at 5, 10 and 20 µg/L. Controls were kept in embryo water only. Behavioral and biochemical parameters were evaluated at 24, 28, 72, and 168 hpf after MZ exposure. The results showed that MZ significantly altered spontaneous movement, escape responses, swimming capacity, and exploratory behavior at all exposure times. However, changes in ROS steady-stead levels and the activity of antioxidant enzymes were observable only at 72 and 168 hpf. In conclusion, behavioral changes occurred earlier than biochemical alterations in zebrafish embryos exposed to MZ, highlighting the potential of behavioral biomarkers as sensitive tools for biomonitoring programs.

A culpa é sempre da mãe? Um olhar sobre a maternidade através da clínica com crianças / ¿La culpa siempre es de la madre? Una mirada hacia la maternidad a través de la clínica con niños / Is it always the mother to blame? A look at maternity through clinical practice with Children / La faute est-elle toujours de la mère? Un regard sur la maternité à travers la clinique de l'enfant

Este artigo pretende discutir questões relativas à maternidade a partir da prática clínica com crianças. Partimos das queixas relacionadas a alguns sintomas infantis que nos chegam para atendimento psicanalítico, para discutir o lugar da mãe na clínica de crianças. Levando em consideração que as mudanças sociohistóricas repercutem nas funções de cuidado dos filhos nas famílias, investigamos de que forma o aumento das possibilidades de destinos para as mulheres repercute nas experiências de maternidade nos dias de hoje. Utilizamos fragmentos de casos clínicos para ilustrar a discussão, evidenciando os desafios de envolver, sem culpabilizar, as mães nos atendimentos de criança

Este artículo pretende discutir cuestiones relativas a la maternidad a partir de la práctica clínica con niños. Partimos de quejas relacionadas a algunos síntomas infantiles que identificamos en el servicio psicoanalítico, para en seguida estudiar el lugar de la madre en la clínica con niños. Teniendo en cuenta que los cambios socioculturales influyen en las funciones de cuidado de los hijos en las familias, investigamos de qué forma el aumento de las posibilidades de destinos para las mujeres en la actualidad repercute en las experiencias de maternidad. Utilizamos fragmentos de casos clínicos para ilustrar la discusión, evidenciando los desafíos de involucrar a la madre en los servicios de niños sin culparla

This article seeks to discuss maternity-related issues based on clinical practice with children. We started by gathering complaints related to certain infant symptoms that we identified at the psychoanalytical service and proceeded to study the place of the mother in clinical practice with children. Considering that sociocultural changes influence childcaring roles in families, we investigated how the current increase of possibilities for women affect maternity experiences. We used excerpts from clinical cases in order to illustrate the discussion, highlighting the challenges of involving the mother when servicing children without blaming her

Cet article se propose de traiter de questions relatives à la maternité, à partir de la pratique clinique auprès d'enfants. Nous partons de plaintes liées à quelques symptômes infantiles qui nous parviennent en thérapie psychanalytique, pour interroger la place de la mère dans la clinique de l'enfant. Tout en prenant en considération que les changements socio-historiques traversent les fonctions de soin vis-à-vis des enfants dans les familles, nous cherchons à comprendre de quelle façon l'augmentation des possibilités de devenirs pour les femmes, se répercute sur les expériences de maternité de nos jours. Des fragments de cas cliniques sont utilisés pour illustrer la discussion et mettent en évidence les enjeux d'impliquer, sans culpabiliser, les mères dans l'accompagnement de leur enfant