Not Only RET but NF1 and Chromosomal Instability Are Seen in Young Patients with Sporadic Medullary Thyroid Carcinoma.

Context: Genetic analysis of sporadic medullary thyroid carcinoma (MTC) has revealed somatic variants in RET, RAS, and occasionally other genes. However, around 20% of patients with sporadic MTC lack a known genetic driver. Objective: To uncover potential new somatic or germline drivers, we analyze a distinct cohort of patients with sporadic, very early-onset, and aggressive MTC. Methods: Germline and somatic DNA exome sequencing was performed in 19 patients, previously tested negative for germline RET variants. Results: Exome sequencing of 19 germline samples confirmed the absence of RET and identified an NF1 pathogenic variant in 1 patient. Somatic sequencing was successful in 15 tumors revealing RET variants in 80%, predominantly p.Met918Thr, which was associated with disease aggressiveness. In RET-negative tumors, pathogenic variants were found in HRAS and NF1. The NF1 germline and somatic variants were observed in a patient without a prior clinical diagnosis of neurofibromatosis type 1, demonstrating that the loss of heterozygosity of NF1 functions as a potential MTC driver. Somatic copy number alterations analysis revealed chromosomal alterations in 53.3% of tumors, predominantly in RET-positive cases, with losses in chromosomes 9 and 22 being the most prevalent. Conclusion: This study reveals that within a cohort of early-onset nonhereditary MTC, RET remains the major driver gene. In RET-negative tumors, NF1 and RAS are drivers of sporadic MTC. In addition, in young patients without a RET germline mutation, a careful clinical evaluation with a consideration of germline NF1 gene analysis is ideal to exclude Neurofibromatosis type 1 (NF1).

Decarbonizing the Transport of Microalgae-based Products - The Role of E-mobility.

BACKGROUND: The decarbonization of road transport is a precondition for achieving carbon neutrality. Battery-electric vehicle technology can make this a reality. In this bias, the objective of the article is to shed light on the ongoing debate about the potentially important role of the adoption of electric vehicles in the transport of microalgae- based products to help them advance to a cleaner life cycle. METHODS: Five routes, including unimodal and multimodal conditions, were defined to assess the carbon emissions of the transport system and, more specifically, of road transport. The headquarters of market-leading microalgae manufacturers were selected as the origin of the routes and, as the destination, regions that sustain them. RESULTS: The results reveal the supremacy of road transport of microalgae-based products using electric vehicles powered by nuclear, hydroelectric, and wind, followed by biomass and photovoltaic energy. They also show that the positive impact of wind, water, and photovoltaic energy on the climate, added to the lower battery charging costs and the greater opportunity to generate revenue from the sale of carbon credits, make their tradeoffs. CONCLUSION: The exquisite results of this study convey key messages to decision-makers and stakeholders about the role of electromobility in building a zero-carbon delivery route.

Fear of public speaking: the effects of a communicational improvement training on physiological parameters and the perception of communication.

Presentations to audiences are often seen as challenging by university students, causing physiological reactivity on cortisol levels and heart rate, for example. Many students perceive that they have difficulties expressing themselves or do not consider themselves to be good communicators. With the thought that efficient communication is able to bring well-being and more confidence, it is understood that a communication skills training could mitigate adverse effects on the body during public speaking. This study aimed to verify whether a communicational improvement training can improve physiological parameters and perception when speaking in public among university students. This was a prospective, blinded, controlled, and randomized trial. Students from colleges and universities were recruited for this study. Invitations were either in person or via social networks and/or e-mail. There were 39 university students who completed the tasks, ages between 18 and 30. There were two groups: the intervention group (IG) and control group (CG). The intervention group participated in a communicational improvement program for six weeks to work on breathing, articulation, speech rhythm, pneumophonoarticulatory coordination, expressiveness and non-verbal communication. Data of heart rate measures, salivary cortisol analysis, self-perception of public speaking, and presentation ratings were collected. The intervention group presented with lower heart rates and cortisol levels, better presentation ratings, and higher self-perception of public speaking than the control group after participating in the communication improvement training. In this study it was possible to verify the efficiency of a communicational improvement training, being able to improve cortisol levels, heart rate, and perception when speaking in public in university students. Based on the results presented, new studies are suggested: (a) measuring the individual effect of the workshops demonstrated here and (b) comparing this type of intervention with other types of treatment used in the health area (medication, psychotherapeutic, holistic, etc.).

Hepatobiliary disease after bone marrow transplant: A cross-sectional study of 377 patients.

BACKGROUND: Bone marrow transplantation (BMT) is a standard treatment for several haematologic conditions. Following BMT, patients may develop hepatobiliary complications that impact morbidity and mortality. The differential diagnosis may include drug-induced liver injury (DILI), sepsis-associated liver injury (SALI), sinusoidal obstruction syndrome (SOS), graft-versus-host disease (GVHD), viral hepatitis, ischaemic hepatitis, and fulminant hepatitis. AIMS: To evaluate the frequency, clinical characteristics, and outcomes of patients with hepatobiliary alterations associated with BMT in a tertiary referral centre. METHODS: This was a cross-sectional study with data collected from the medical records of patients undergoing BMT between January 2017 and June 2022. We diagnosed hepatobiliary complications based on established criteria. RESULTS: We included 377 patients; 55.7% had hepatobiliary complications. Female gender, pre-BMT hepatobiliary alteration, and haploidentical allogeneic transplantation were associated with increased risk with odds ratios (OR) of 1.8 (p = 0.005), 1.72 (p = 0.013) and 3.25 (p = 0.003), respectively. Patients with hepatobiliary complications spent longer in the hospital than those without (27.7 × 19.3 days, respectively; p < 0.001). Among 210 patients with hepatobiliary complications, 28 died compared to 5 of 167 without complications (OR 4.98; p = 0.001). CONCLUSIONS: Hepatobiliary complications are frequent in patients undergoing BMT. There is a greater risk of their occurrence in women, people with pre-BMT liver alterations, and in haploidentical transplants. The occurrence of these complications increases the length of stay and is associated with a higher risk of death.

Evaluation of silver nanoparticle-impregnated PMMA loaded with vancomycin or gentamicin against bacterial biofilm formation.

INTRODUCTION: Bone cement containing vancomycin or gentamicin is a therapeutic strategy for combating orthopedic infections: however, the activity of these antibiotics is narrow. Silver nanoparticles (AgNPs) are nanocomponents with a wide spectrum, including multidrug-resistant bacteria. In the present study, we aimed to evaluate the effect of AgNP-loaded polymethylmethacrylate (PMMA) on biofilm formation by Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus epidermidis. METHODS: The effect of AgNP-loaded PMMA with and without vancomycin or gentamicin on biofilm production was quantitatively analyzed. S. aureus, E. coli, P. aeruginosa, and S. epidermidis were included as biofilm-producing microorganisms in the in vitro model. RESULTS: AgNP-loaded PMMA with antibiotics reduced the number of colony-forming units (CFUs; p<0.001). However, AgNP-loaded PMMA alone did not significantly reduce biofilm formation. CONCLUSION: Our study demonstrated the potential of AgNP-loaded PMMA. Notably, we observed that AgNP-loaded PMMA containing vancomycin or gentamycin exhibited significantly superior efficacy, with satisfactory activity against most biofilm-forming microbial agents examined.

CRISPR/Cas-9 vector system: targets UL-39 and inhibits Simplexvirus humanalpha1 (HSV-1) replication in vitro.

Simplexvirus humanalpha1 (HSV-1) affects approximately 67% of the world's population. Here, we sought to use the CRISPR / Cas9 system with the UL39 target, essential for virus replication. The sgRNA sequence was inserted into the plasmid (PX459-UL39). Vero cells were transfected with PX459-UL39, and inhibition of viral replication was assessed 24 and 48 hours later using plaque assays and fluorescence and qPCR. Fluorescence analyses revealed the presence of anti-HSV-1 CRISPR/Cas9 within Vero cells, and qPCR showed that the viral load decreased by> 95% of cells transfected with anti-HSV-1 CRISPR / Cas 9. Our data demonstrate the usefulness of the PX459-UL39 to inhibit HSV-1 infection.

Physical conditioning in children and adolescents with cerebral palsy: Systematic review and meta-analysis.

INTRODUCTION: Cerebral palsy (CP) can be described as a group of permanent non-progressive disorders that occur in the developing fetal or infant brain. Studies have shown that children and adolescents with CP have low cardiorespiratory fitness and higher energy expenditure during daily activities when compared to typical children. Therefore, interventions focused on the physical conditioning of this population could be critical. OBJECTIVE: To evaluate the effect of physical conditioning training on distance walked and maximum oxygen consumption (VO2 max) in individuals with CP, through a systematic review. METHOD: Two independent researchers performed systematic searches in the PUBMED, SciELO, PEDro, ERIC, and Cochrane databases using the search terms "physical fitness", "aerobic training" or "endurance" combined with "cerebral palsy". INCLUSION CRITERIA: (1) population (children and adolescents with CP between 5 and 18 years old); (2) type of study (experimental studies); (3) type of intervention (physical conditioning protocols); (4) outcome (distance walked in the 6 min walking test - 6MWT, and VO2 max). RESULTS: 386 studies were identified and 5 articles were considered eligible. After physical conditioning training, there was an increase of 46.34 m (p = 0.07) and 5.93. ml. kg-1. min -1 (p < 0.001) in the 6MWT and VO2 max, respectively. CONCLUSION: Physical conditioning training appears to be clinically beneficial to the cardiorespiratory fitness of children and adolescents with CP.

A systematic review of clinical trials for gene therapies for ß-hemoglobinopathy around the world.

BACKGROUND AIMS: Amidst the success of cell therapy for the treatment of onco-hematological diseases, the first recently Food and Drug Administration-approved gene therapy product for patients with transfusion-dependent ß-thalassemia (TDT) indicates the feasibility of gene therapy as curative for genetic hematologic disorders. This work analyzed the current-world scenario of clinical trials involving gene therapy for ß-hemoglobinopathies. METHODS: Eighteen trials for patients with sickle cell disease (SCD) and 24 for patients with TDT were analyzed. RESULTS: Most are phase 1 and 2 trials, funded by the industry and are currently recruiting volunteers. Treatment strategies for both diseases are fetal hemoglobin induction (52.4%); addition of wild-type or therapeutic ß-globin gene (38.1%) and correction of mutations (9,5%). Gene editing (52.4%) and gene addition (40.5%) are the two most used techniques. The United States and France are the countries with the greatest number of clinical trials centers for SCD, with 83.1% and 4.2%, respectively. The United States (41.1%), China (26%) and Italy (6.8%) lead TDT trials centers. CONCLUSIONS: Geographic trial concentration indicates the high costs of this technology, logistical issues and social challenges that need to be overcome for gene therapy to reach low- and middle-income countries where SCD and TDT are prevalent and where they most impact the patient's health.

Short-Term Intake of Theobroma grandiflorum Juice Fermented with Lacticaseibacillus rhamnosus ATCC 9595 Amended the Outcome of Endotoxemia Induced by Lipopolysaccharide.

Endotoxemia is a condition caused by increasing levels of lipopolysaccharide (LPS) characterized by an impaired systemic response that causes multiple organ dysfunction. Lacticaseibacillus rhamnosus ATCC 9595 is a strain with probiotic potential which shows immunomodulatory properties. The incorporation of this bacterium in food rich in bioactive compounds, such as cupuaçu juice (Theobroma grandiflorum), could result in a product with interesting health properties. This work evaluated the effects of the oral administration of cupuaçu juice fermented with L. rhamnosus on the outcome of LPS-induced endotoxemia in mice. C57BL/6 mice (12/group) received oral doses (100 µL) of saline solution and unfermented or fermented cupuaçu juice (108 CFU/mL). After 5 days, the endotoxemia was induced by an intraperitoneal injection of LPS (10 mg/kg). The endotoxemia severity was evaluated daily using a score based on grooming behavior, mobility, presence of piloerection, and weeping eyes. After 6 h and 120 h, the mice (6/group) were euthanized for analysis of cell counts (in peritoneal lavage and serum) and organ weight. L. rhamnosus grew in cupuaçu juice and produced organic acids without the need for supplementation. The bacteria counts were stable in the juice during storage at 4 °C for 28 days. The fermentation with L. rhamnosus ATCC 9595 changed the metabolites profile of cupuaçu juice due to the biotransformation and enhancement of some compounds. In general, the administration of L. rhamnosus-fermented juice allowed a significant improvement in several characteristics of endotoxemic status (weight loss, hypothermia, severity index, cell migration). In addition, treatment with fermented juice significantly reduced the weight of the spleen, liver, intestine, and kidneys compared to the saline-treated endotoxemic group. Taken together, our data show that short-term intake therapy of cupuaçu juice fermented with L. rhamnosus ATCC 9595 can reduce systemic inflammation in an experimental model of LPS-induced endotoxemia in mice.

Benchmarking the Effects on Human-Exoskeleton Interaction of Trajectory, Admittance and EMG-Triggered Exoskeleton Movement Control.

Nowadays, robotic technology for gait training is becoming a common tool in rehabilitation hospitals. However, its effectiveness is still controversial. Traditional control strategies do not adequately integrate human intention and interaction and little is known regarding the impact of exoskeleton control strategies on muscle coordination, physical effort, and user acceptance. In this article, we benchmarked three types of exoskeleton control strategies in a sample of seven healthy volunteers: trajectory assistance (TC), compliant assistance (AC), and compliant assistance with EMG-Onset stepping control (OC), which allows the user to decide when to take a step during the walking cycle. This exploratory study was conducted within the EUROBENCH project facility. Experimental procedures and data analysis were conducted following EUROBENCH's protocols. Specifically, exoskeleton kinematics, muscle activation, heart and breathing rates, skin conductance, as well as user-perceived effort were analyzed. Our results show that the OC controller showed robust performance in detecting stepping intention even using a corrupt EMG acquisition channel. The AC and OC controllers resulted in similar kinematic alterations compared to the TC controller. Muscle synergies remained similar to the synergies found in the literature, although some changes in muscle contribution were found, as well as an overall increase in agonist-antagonist co-contraction. The OC condition led to the decreased mean duration of activation of synergies. These differences were not reflected in the overall physiological impact of walking or subjective perception. We conclude that, although the AC and OC walking conditions allowed the users to modulate their walking pattern, the application of these two controllers did not translate into significant changes in the overall physiological cost of walking nor the perceived experience of use. Nonetheless, results suggest that both AC and OC controllers are potentially interesting approaches that can be explored as gait rehabilitation tools. Furthermore, the INTENTION project is, to our knowledge, the first study to benchmark the effects on human-exoskeleton interaction of three different exoskeleton controllers, including a new EMG-based controller designed by us and never tested in previous studies, which has made it possible to provide valuable third-party feedback on the use of the EUROBENCH facility and testbed, enriching the apprenticeship of the project consortium and contributing to the scientific community.

Detecção de flavivirus e alphavirus em mosquitos (diptera: culicidae) capturados em região próxima à Três Lagoas ­ Ms / Detection of flavivirus and alphavirus in mosquitoes (diptera: culicidae) captured in a region close to Tres Lagoas ­ Ms / Detección de flavivirus y alphavirus en mosquitos (diptera: culicidae) capturados en una región cercana a Tres Lagoas ­ Ms

Introdução: Arbovírus são causadores de doenças humanas, sendo que mudança ecológicas e aumento do contato humano-vetor aumenta a possibilidade de surtos. Objetivo: Detectar, identificar e caracterizar arbovírus presentes em mosquitos vetores capturados em regiões de mata próximas a Três Lagoas, MS. Metodologia: Mosquitos foram capturados utilizando armadilhas de luz em regiões de mata circunvizinha a Três Lagoas. Os mosquitos capturados foram classificados por gênero (chave morfológica) e agrupados em pools com até 20 espécimes, e utilizados através da reação de RT-PCR com posterior sequenciamento e análise filogenética. Resultados: Foram capturados 851 dos gêneros: Culex spp. (11 pools); Aedes spp. (13 pools); Haemagogus spp. (7 pools) e outros gêneros não identificados. Sequencias de vírus Dengue (DENV) foram amplificadas de 2/13 (15,38%) pools de Aedes spp. e uma sequência de vírus Mayaro (MAYV) 1/7 (7,7%) foi amplificada de pools de Haemagogus spp. As análises filogenéticas mostraram que as sequências de DENV agrupava-se no clado de DENV1 e DENV2. A sequência de MAYV agrupou-se junto a sequências de amostras de infecções humana por MAYV do grupo L. Conclusão: Estes resultados reforçam a circulação de DENV, que é causador de surtos anuais de doenças febris agudas no município, e detecção, por primeira vez na região, a circulação de MAYV, reforçando a necessidade de monitoramento viral constante nessa região.

Introduction: Arboviruses cause human diseases, and ecological changes and increased human-vector contact increase the possibility of outbreaks. Objective: To detect, identify and characterize arboviruses present in mosquito vectors captured in forest regions close to Tres Lagoas, MS. Methodology: Mosquitoes were captured using light traps in forest regions surrounding Tres Lagoas. The captured mosquitoes were classified by gender (morphological key) and grouped into pools with up to 20 specimens and used through the RT-PCR reaction with subsequent sequencing and phylogenetic analysis. Results: 851 of the genera were captured: Culex spp. (11 pools); Aedes spp. (13 pools); Haemagogus spp. (7 pools) and other unidentified genera. Dengue virus (DENV) sequences were amplified from 2/13 (15.38%) pools of Aedes spp. and a Mayaro virus (MAYV) sequence 1/7 (7.7%) were amplified from pools of Haemagogus spp. Phylogenetic analyzes showed that one of the DENV sequences clustered in the DENV1 and DENV2 clade. The MAYV sequence was grouped together with sequences from samples of human MAYV infections of the L group. Conclusion: These results reinforce the circulation of DENV, which causes annual outbreaks of acute febrile illnesses in the municipality, and detection, for the first time in the region, the circulation of MAYV, reinforcing the need for constant viral monitoring in this region.

Introducción: Los arbovirus causan enfermedades humanas, y los cambios ecológicos y el mayor contacto humano-vector aumentan la posibilidad de brotes. Objetivo: Detectar, identificar y caracterizar arbovirus presentes en mosquitos vectores capturados en regiones de selva próximas a Tres Lagoas, MS. Metodología: Los mosquitos fueron capturados utilizando trampas de luz en las regiones forestales que rodean Tres Lagoas. Los mosquitos capturados fueron clasificados por género (clave morfológica) y agrupados en pools de hasta 20 ejemplares, y utilizados mediante la reacción RT-PCR con posterior secuenciación y análisis filogenético. Resultados: Se capturaron 851 de los géneros: Culex spp. (11 pools); Aedes spp. (13 pools); Haemagogus spp. (7 pools) y otros géneros no identificados. Las secuencias del virus del dengue (DENV) se amplificaron a partir de 2/13 (15,38 %) grupos de Aedes spp. y una secuencia de virus Mayaro (MAYV) 1/7 (7,7%) de pools de Haemagogus spp. Los análisis filogenéticos mostraron que una de las secuencias de DENV se agrupaba en el clado DENV1 y DENV2. La secuencia de MAYV se agrupó con secuencias de muestras de infecciones humanas de MAYV del grupo L. Conclusión: Estos resultados refuerzan la circulación de DENV, causante de brotes anuales de enfermedades febriles agudas en el municipio, y la detección, por primera vez en la región, la circulación de MAYV, reforzando la necesidad de un monitoreo viral constante en esta región.

Homeostasis Maintenance in Plasmodium-Infected Placentas: Is There a Role for Placental Autophagy During Malaria in Pregnancy?

Malaria represents a significant public health burden to populations living in developing countries. The disease takes a relevant toll on pregnant women, who are more prone to developing severe clinical manifestations. Inflammation triggered in response to P. falciparum sequestration inside the placenta leads to physiological and structural changes in the organ, reflecting locally disrupted homeostasis. Altogether, these events have been associated with poor gestational outcomes, such as intrauterine growth restriction and premature delivery, contributing to the parturition of thousands of African children with low birth weight. Despite significant advances in the field, the molecular mechanisms that govern these outcomes are still poorly understood. Herein, we discuss the idea of how some housekeeping molecular mechanisms, such as those related to autophagy, might be intertwined with the outcomes of malaria in pregnancy. We contextualize previous findings suggesting that placental autophagy is dysregulated in P. falciparum-infected pregnant women with complementary research describing the importance of autophagy in healthy pregnancies. Since the functional role of autophagy in pregnancy outcomes is still unclear, we hypothesize that autophagy might be essential for circumventing inflammation-induced stress in the placenta, acting as a cytoprotective mechanism that attempts to ensure local homeostasis and better gestational prognosis in women with malaria in pregnancy.

Systematic Review of Available CAR-T Cell Trials around the World.

In this systematic review, we foresee what could be the approved scenario in the next few years for CAR-T cell therapies directed against hematological and solid tumor malignancies. China and the USA are the leading regions in numbers of clinical studies involving CAR-T. Hematological antigens CD19 and BCMA are the most targeted, followed by mesothelin, GPC3, CEA, MUC1, HER2, and EGFR for solid tumors. Most CAR constructs are second-generation, although third and fourth generations are being largely explored. Moreover, the benefit of combining CAR-T treatment with immune checkpoint inhibitors and other drugs is also being assessed. Data regarding product formulation and administration, such as cell phenotype, transfection technique, and cell dosage, are scarce and could not be retrieved. Better tracking of trials' status and results on the ClinicalTrials.gov database should aid in a more concise and general view of the ongoing clinical trials involving CAR-T cell therapy.

COVID-19 impact on anxiety and depression in head and neck cancer patients: A cross-sectional study.

OBJECTIVE: To evaluate whether the coronavirus disease 2019 has increased anxiety, depression, and distress levels in head and neck cancer (HNC) patients undergoing radiotherapy (RT). METHODS: In this cross-sectional study, RT-HNC patients were surveyed using the Hospital Anxiety and Depression Scale (HADS) for anxiety and depression and the distress thermometer (DT) for distress. HADS scores were compared with data pre-COVID-19. Additionally, we evaluated the COVID-19 impact on daily routines, treatment, and cancer care through a questionnaire. RESULTS: Fifty patients were included. The HADS mean score and estimated rates were 4.34 (±4.06)/22% for anxiety and 5.08 (±4.82)/22% for depression; in comparison, our historical control had 4.04 (±3.59)/20% for anxiety (p = .79) and 4.03 (±3.62)/17% for depression (p = .49). Mean DT score was 3.68 (±2.77). Responders were aware of COVID-19, afraid of having medical complications, believed it was life-threatening, did not miss appointments, believed their treatment was not impacted, and felt safe at the hospital amid the pandemic. CONCLUSION: This study suggests that anxiety, depression, and distress levels found in RT-HNC patients did not increase during the pandemic. Patients were afraid of being infected by COVID-19; however, they complied with their cancer treatment.

Primary Biliary Cholangitis: A Case of Underdiagnosis.

Primary biliary cholangitis is a rare autoimmune cholestatic disease with a variable clinical course. Its etiopathogenesis has not been completely clarified. It predominantly affects women and often progresses to liver cirrhosis. It may be asymptomatic or symptomatic with hepatic or extrahepatic manifestations. If its diagnosis and treatment are made early, the progression to cirrhosis and liver failure can be prevented. We describe a clinical case of a 63-year-old woman, with no history of hepatotoxic drugs, who presented an analytical pattern of cytocholestasis with more than one decade of evolution.

Does Subcrestal Position Affect Insertion Torque of Different Implant Designs at Different Bone Densities? An In Vitro Model Study.

PURPOSE: To evaluate the insertion torque at the equicrestal and subcrestal positions of three implant designs in synthetic polyurethane blocks simulating different bone conditions. MATERIALS AND METHODS: Overall, 72 implants with three different designs (two conical and one cylindrical) were inserted at equicrestal and subcrestal positions (-2 mm) into polyurethane blocks simulating the anterior (0.32 g/cm3) and posterior (0.48 g/cm3) regions of the maxilla. The final insertion torque of all implants was measured using a digital torque meter, and the results were evaluated and statistically analyzed. RESULTS: For all implant systems, insertion torque decreased significantly when placed at the subcrestal position (P < .05), except for cylindrical implants placed in the bone blocks with high cancellous density. The implants with higher body conicity and square threads had higher insertion torque values than the cylindrical and conical implants with a lower body conical angle and trapezoidal threads. The implants inserted into higher-density blocks showed greater stability. CONCLUSION: The different implant systems lost insertion torque in a nonuniform manner at the subcrestal position and varied according to bone density. Implant design significantly influenced the initial stability at the equicrestal and subcrestal positions, especially in lower-density bone.

In Search of an Ideal CAR-T Cell Antigen Target.

Chimeric antigen receptor T (CAR-T) cells are proving their value in hematological cancers such as B cell acute lymphoid leukemia (B-ALL). This success is in great part due to the chosen target antigen, the lineage marker CD19, that is expressed by leukemia blasts and B lymphocytes, which are not crucial. For solid tumors, the challenge is greater because antigen expression is highly heterogeneous within the tumor and even an efficient CAR-T strategy would not kill all tumor cells. Also, many antigens are shared between solid tumors and healthy cells, causing off-target cell lysis and dangerous collateral damage. New antigen sources are emerging as targets, such as viruses, endogenous viruses, and immune checkpoint molecules. New technologies are in search of the ideal target, with antigen combinations the leading candidates.

CRISPR / Cas-9 Vector System: Targets UL-39 and Inhibits HSV-1 Replication in Vitro

HSV-1 affects approximately 67% of the world population. Here, we sought to use the CRISPR / Cas9 system with the UL39 target, essential for virus replication. The sgRNA sequence was inserted into plasmid (PX459-UL39). Vero cells were transfected with PX459-UL39, and inhibition of viral replication was assessed 24 and 48 hours later using plaque assays and fluorescence and qPCR. Fluorescence analyzes revealed the presence of anti-HSV-1 CRISPR/Cas9 within Vero cells, and qPCR showed that the viral load decreased by> 95% of cells transfected with anti-HSV-1 CRISPR / Cas9. Our data demonstrate the usefulness of the PX459-UL39 to inhibit HSV-1 infection.