The Use of Flavylium Salts as Dynamic Inhibitor Moieties for Human Cb5R.

Cytochrome b5 reductase (Cb5R) is a flavoprotein that participates in the reduction of multiple biological redox partners. Co-localization of this protein with nitric oxide sources has been observed in neurons. In addition, the generation of superoxide anion radical by Cb5R has been observed. A search for specific inhibitors of Cb5R to understand the role of this protein in these new functions has been initiated. Previous studies have shown the ability of different flavonoids to inhibit Cb5R. Anthocyanins are a subgroup of flavonoids responsible for most red and blue colors found in flowers and fruits. Although usually represented by the flavylium cation form, these species are only stable at rather acidic pH values (pH ≤ 1). At higher pH values, the flavylium cation is involved in a dynamic reaction network comprising different neutral species with the potential ability to inhibit the activities of Cb5R. This study aims to provide insights into the molecular mechanism of interaction between flavonoids and Cb5R using flavylium salts as dynamic inhibitors. The outcome of this study might lead to the design of improved specific enzyme inhibitors in the future.

Identification of a truncated ß1-chimaerin variant that inactivates nuclear Rac1.

ß1-chimaerin belongs to the chimaerin family of GTPase-activating proteins (GAPs) and is encoded by the CHN2 gene, which also encodes the ß2- and ß3-chimaerin isoforms. All chimaerin isoforms have a C1 domain that binds diacylglycerol as well as tumor-promoting phorbol esters and a catalytic GAP domain that inactivates the small GTPase Rac. Nuclear Rac has emerged as a key regulator of various cell functions, including cell division, and has a pathological role by promoting tumorigenesis and metastasis. However, how nuclear Rac is regulated has not been fully addressed. Here, using several approaches, including siRNA-mediated gene silencing, confocal microscopy, and subcellular fractionation, we identified a nuclear variant of ß1-chimaerin, ß1-Δ7p-chimaerin, that participates in the regulation of nuclear Rac1. We show that ß1-Δ7p-chimaerin is a truncated variant generated by alternative splicing at a cryptic splice site in exon 7. We found that, unlike other chimaerin isoforms, ß1-Δ7p-chimaerin lacks a functional C1 domain and is not regulated by diacylglycerol. We found that ß1-Δ7p-chimaerin localizes to the nucleus via a nuclear localization signal in its N terminus. We also identified a key nuclear export signal in ß1-chimaerin that is absent in ß1-Δ7p-chimaerin, causing nuclear retention of this truncated variant. Functionally analyses revealed that ß1-Δ7p-chimaerin inactivates nuclear Rac and negatively regulates the cell cycle. Our results provide important insights into the diversity of chimaerin Rac-GAP regulation and function and highlight a potential mechanism of nuclear Rac inactivation that may play significant roles in pathologies such as cancer.

Hammering home the sickle: an instructive case of endobronchial anaplastic large cell lymphoma.

It is extremely unusual for primary pulmonary lymphoma, an uncommon occurrence in any form, to be of T-cell origin and to manifest as an endobronchial lesion. Each of these characteristics is rare individually, so cases that combine them are exceptional. We report a patient in whom primary pulmonary anaplastic large cell lymphoma, a T-cell neoplasm, presented with obstruction of the left upper lobe bronchus, resulting in left upper lobe atelectasis and creating the radiographic luftsichel sign. We briefly discuss anaplastic large cell lymphoma as a whole, place our case in the context of previously published literature on endobronchial anaplastic large cell lymphoma, and review the genesis of the luftsichel sign.

Three's company: A woman with rash, uterine mass and cystic lungs.

Cystic lung disease encompasses a wide variety of clinical entities, the diagnosis of which is sometimes straightforward and other times obscure.  To narrow the list of possibilities, it behooves the physician to consider the context in which the cystic lung disease is uncovered. Clues to the diagnosis might be provided by findings that are not initially obvious and are not located in the thorax. We describe an instructive case of a woman with cystic lungs detected during a search for malignancy prompted by a diagnosis of dermatomyositis. Malignancy was indeed uncovered in the form of endometrial carcinoma, the management of which eventually also established the etiology of cystic lung disease. In the discussion we attempt to connect the patient's autoimmune disease, uterine cancer, and lung cysts. The potential interplay among these three components of her presentation makes for intriguing mechanistic speculation.

Labiaplasty: A 24-Month Experience in 58 Patients: Outcomes and Statistical Analysis.

BACKGROUND: Because there has been a great raise in the interest of this type of surgery in our area, we decided to study if there were any differences among our patients and also to review our results. OBJECTIVE: The purpose of this study was to review our experience with labiaplasty. We assessed surgical complications and quality outcomes and compared surgical results according to maternal parity. METHODS: We conducted a retrospective chart review of all cases of primary or secondary labia minora reduction surgery performed by the same surgeon from January 2014 to December 2015. We recorded patient demographics, sexual activity, parity, and presurgical and postsurgical quality of life, as well as surgical technique and complications. RESULTS: During the study period, we performed 58 labia minora reduction procedures. The average patient age was 32.16 years. Among the patients, 50% were single, 65.52% sexually active, and 62.07% nulliparous. A wedge excision was performed in 75.8% of cases. A complication occurred in 12.06% of cases, usually owing to underresection. The surgical experience was rated very good or excellent by 96.55% of patients. Women with children reported greater satisfaction with their surgical outcome on our postoperative questionnaire. We identified no statistically significant outcome differences according to surgical technique. CONCLUSIONS: Labiaplasty is safe and shows high rates of overall satisfaction. Women with children reported greater postoperative satisfaction than women who had never given birth.

Charge-Carrier Dynamics in 2D Hybrid Metal-Halide Perovskites.

Hybrid metal-halide perovskites are promising new materials for use in solar cells; however, their chemical stability in the presence of moisture remains a significant drawback. Quasi two-dimensional (2D) perovskites that incorporate hydrophobic organic interlayers offer improved resistance to degradation by moisture, currently still at the cost of overall cell efficiency. To elucidate the factors affecting the optoelectronic properties of these materials, we have investigated the charge transport properties and crystallographic orientation of mixed methylammonium (MA)-phenylethylammonium (PEA) lead iodide thin films as a function of the MA-to-PEA ratio and, thus, the thickness of the "encapsulated" MA lead-halide layers. We find that monomolecular charge-carrier recombination rates first decrease with increasing PEA fraction, most likely as a result of trap passivation, but then increase significantly as excitonic effects begin to dominate for thin confined layers. Bimolecular and Auger recombination rate constants are found to be sensitive to changes in electronic confinement, which alters the density of states for electronic transitions. We demonstrate that effective charge-carrier mobilities remain remarkably high (near 10 cm2V-1s-1) for intermediate PEA content and are enhanced for preferential orientation of the conducting lead iodide layers along the probing electric field. The trade-off between trap reduction, electronic confinement, and layer orientation leads to calculated charge-carrier diffusion lengths reaching a maximum of 2.5 µm for intermediate PEA content (50%).

Five-Year Outcomes of Breast Augmentation with Form-Stable Implants: Periareolar vs Transaxillary.

BACKGROUND: Form-stable 410 implants have the potential advantage of maintaining their anatomic form thanks to the cohesiveness of the gel. Furthermore, Biocell texturing appears to maximize adhesion and to allow for implant immobility. OBJECTIVES: To compare the rate of reoperations for transaxillary and periareolar approaches for breast augmentation. METHODS: This retrospective study consisted of 373 patients with a 5-year follow up. Patient demographics, self-perception and esteem, surgical technique, and implant characteristics were documented. The reasons for reoperation for both approaches were reviewed. RESULTS: Transaxillary breast augmentation was used in 302 patients (81%) and periareolar breast augmentation in 71 patients (19%). In the axillary group, 210 had subfascial placement (69.5%), and 92 patients had submuscular placement (30.5%). In the nipple-areolar complex group, 50 were subfascial (70.4%), and 21 were submuscular (29.6%). The reoperation rate for the patients operated on during this time and followed for 5 years was 11% (8 patients) for the nipple-areolar complex approach and 8.3% (25 patients) in the axillary group. Capsular contracture grade III or IV were the main causes for reoperation for any technique (4.2% nipple-areolar complex vs 3.3% axillary). Other reasons were implant rupture, seroma, infection, implant malrotation, implant malposition, and rippling. CONCLUSIONS: The rate of reoperations was similar to those described in the literature for this type of implant. There were no statistically significant differences between the various techniques, although the reoperation rate was significantly higher when a periareolar subfascial technique was used. LEVEL OF EVIDENCE: 3 Therapeutic.

Solution Synthesis Approach to Colloidal Cesium Lead Halide Perovskite Nanoplatelets with Monolayer-Level Thickness Control.

We report a colloidal synthesis approach to CsPbBr3 nanoplatelets (NPLs). The nucleation and growth of the platelets, which takes place at room temperature, is triggered by the injection of acetone in a mixture of precursors that would remain unreactive otherwise. The low growth temperature enables the control of the plate thickness, which can be precisely tuned from 3 to 5 monolayers. The strong two-dimensional confinement of the carriers at such small vertical sizes is responsible for a narrow PL, strong excitonic absorption, and a blue shift of the optical band gap by more than 0.47 eV compared to that of bulk CsPbBr3. We also show that the composition of the NPLs can be varied all the way to CsPbBr3 or CsPbI3 by anion exchange, with preservation of the size and shape of the starting particles. The blue fluorescent CsPbCl3 NPLs represent a new member of the scarcely populated group of blue-emitting colloidal nanocrystals. The exciton dynamics were found to be independent of the extent of 2D confinement in these platelets, and this was supported by band structure calculations.

Phase II clinical experience and long-term follow-up using the next-generation single-incision platform FMX314.

BACKGROUND: Single-incision minimally invasive surgery has previously been associated with incisions 2.0-3.0 cm in length. We present a novel single-incision surgical platform compatible for insertion through a standard 15-mm trocar we previously described in six patients with short-term follow-up data. The objective of this phase II study was to evaluate the safety and feasibility of the platform in a larger collective and to evaluate 1-year follow-up data of the phase I trial. METHODS: The technology features a multiple-use introducer, accommodating the articulating instruments, and is inserted through a 15-mm laparoscopic trocar. Cholecystectomy is performed through an umbilical incision. A prospective feasibility study was performed at a single center. Inclusion criteria were age of 18-75 years and biliary colic, exclusion criteria were acute cholecystitis, dilation of the biliary tree, severe coagulopathy, BMI > 40 kg/m(2), or choledocholithiasis. Endpoints included complications, length of stay, pain medication, cosmetic results, and the presence of hernia. RESULTS: Twenty-seven patients (23 females; phase I: 6 patients, phase II: 21 patients) with an average age of 41.7 years and BMI 26.6 kg/m(2) were recruited for the study. Umbilical incision length did not exceed 15 mm. There were no intraoperative complications. Average OR time decreased from 91 min for the first six cases to 56 min for the last six cases. Average length of stay was 7.8 h. Pain control was achieved with diclofenac for no more than 7 days. All patients had no adverse events at 5-month follow-up, and all phase I patients had no adverse events nor evidence of umbilical hernia at 1 year. CONCLUSION: This study demonstrates that single-incision cholecystectomy with the platform is feasible, safe, and reproducible in a larger patient population. Long-term follow-up showed no hernias or other adverse events. Further studies will be needed to evaluate longer-term hernia rates.

Physicians' information to patients and prescription of the emergency contraceptive pill according to their personal experience of using the method and perception of its mechanism of action.

OBJECTIVES: The aim of the study was to evaluate the association between physicians' understanding of the mechanism of action of the emergency contraceptive pill (ECP), their personal use of it, and their practice in informing their patients about the method and in prescribing it. METHODS: The study was carried out in a sample of 3337 obstetrician-gynaecologists who responded to a mailed questionnaire. Bivariate analysis was used to test the association between physicians' personal use of the ECP, their understanding of its mechanism of action, and their practice in informing their patients about the method and in prescribing it. Multiple Poisson regression analysis was carried out to identify variables independently associated with the two dependent variables. RESULTS: Multiple regression analysis showed that the percentage of physicians who had informed their patients about the ECP was significantly lower among those who had needed it themselves but had not used it and among those living in the northeast of Brazil. A significantly higher percentage of female than male physicians had provided information on the ECP. The percentage of physicians who had prescribed the ECP was significantly lower among those who had needed it themselves but had not used it and among those who believed that it caused a mini-abortion. The proportion of physicians who had ever-prescribed the ECP was greater among those who worked exclusively in private practice and among those who worked in a state capital. CONCLUSIONS: The misconception that emergency contraception could cause a mini-abortion was associated with its denial to potential users, while physicians' personal experience of needing to use it favoured the likelihood of their informing potential users about it and prescribing it.

Foundational features of indigenous pregnancy care: Lessons learned from indigenous pregnancy care providers.

PROBLEM: Indigenous populations experience higher odds of poor maternal and infant health outcomes than non-Hispanic White mothers yet have lower odds of receiving adequate prenatal care. BACKGROUND: Many Indigenous communities rely on modern Western medical institutions to provide pregnancy related health care. These systems were not developed with or for Indigenous communities and often fail to meet the needs of Indigenous pregnant patients. Offering culturally congruent models of care may increase prenatal care utilization. QUESTION, HYPOTHESIS OR AIM: This paper used qualitative inquiry to identify Indigenous approaches to caring for pregnancy. METHODS: Our team conducted 16 semi-structured individual interviews and one group interview with a total of 19 respondents. To arrive at thematic categories, the research team engaged in a modified pile sorting technique. The final set of categories, along with sub-themes, descriptions and example quotes, were sent to interviewees for approval. FINDINGS: Ten Foundational Features of Indigenous Pregnancy Care were identified. These covered themes related to Indigenous cultural practices, relationships, Indigenous sovereignty, local Indigenous community, full spectrum care, wholistic care, birthing person's wisdom, power and autonomy, flexibility, historical trauma, and cultural awareness. DISCUSSION: Modern midwifery care delivered by Indigenous practitioners may partially bridge the cultural gap; however, intentional effort is needed to integrate Indigenous ways into medical doctor practice models and facilities. CONCLUSION: This paper identifies ten foundational features of Indigenous pregnancy care and demonstrates the importance of recognizing the effects of trauma and providing opportunities for healing, upholding sovereignty, and centering relationships when caring for Indigenous pregnancies.

The Great Italian Masquerade.

Tuberculosis and sarcoidosis are both granulomatous diseases centered on the lung but capable of myriad extrapulmonary manifestations. Because of substantial similarity in their presentations, these two entities can be notoriously challenging to differentiate. This can be particularly true of countries in which tuberculosis is rarely encountered because of a reflexive tendency to ascribe granulomatous inflammation in the lung to sarcoidosis, especially if the granulomas are non-necrotizing. However, as our case from a non-endemic country reminds, sarcoidosis can be comfortably diagnosed only after convincing exclusion of infectious causes of granulomas. Distinguishing these two diseases is of utmost importance as, despite their overlapping presentations, they have completely non-overlapping treatments which can lead to harm if erroneously applied. At the end of our discussion, we summarize the clinical features favoring one diagnosis over the other.

Continuation Rates of the 52-mg Levonorgestrel-releasing Intrauterine System according to the Primary Reason for its Use.

OBJECTIVE: To evaluate whether continuation rates with the 52-mg levonorgestrel-releasing intrauterine system (LNG-IUS) up to 5 years after placement differed between women using the method exclusively for contraception and those using the device for medical reasons alone. METHODS: A retrospective cohort study was conducted in a family planning clinic with 5,034 LNG-IUS users: 4,287 using the method exclusively for contraception and 747 for medical reasons alone. The continuation rate at 1 to 5 years of use was calculated by life table analysis. RESULTS: Initially, the continuation rate was significantly higher in the contraception group: 85.8 versus 83.4 and 77.4 versus 76.0 per 100 women-years in the 1st and 2nd years of use, respectively. There were more discontinuations due to bleeding/spotting in the medical reasons group in the first two years. The discontinuation rate according to reason for use was not significantly different from the third to the fifth year of use. No women discontinued due to amenorrhea in either group. CONCLUSION: The continuation rate was significantly higher in the contraception group in the first two years of use. Amenorrhea was not a reason for discontinuation in either group, suggesting that counselling in this respect was adequate. Nevertheless, counselling could perhaps have been better with regards to the expected long period of bleeding and spotting in the first two years after placement.

OBJETIVO: Avaliar a taxa de continuação até 5 anos de uso do sistema intrauterino liberador de 52-mg levonorgestrel por dia (SIU LNG) -IUS) é diferente entre mulheres que o usam exclusivamente como anticoncepcional que entre as que usam exclusivamente por razões médicas. MéTODOS: Estudo retrospectivo realizado em uma clínica de Planejamento Familiar 5.034 usuárias de SIU LNG, 4.287 que optaram pelo método apenas como anticoncepcional e 747 que o usavam somente por razoes médicas. A taxa de continuação de um até cinco ano foi calculada por meio de análise de tabela de vida RESULTADOS: No início a taxa de continuação foi significativamente maior no grupo da anticoncepção: 85,8 versus 83,4 e 77,4 versus 76,0 por 100 anos-mulher no 1° e 2° ano de uso, respectivamente. Houve mais descontinuações por sangrado-manchado no grupo de razões médicas nos dos primeiros anos. A taxa de continuação não foi significativamente diferente desde o terceiro até o quinto ano de uso. Nenhuma mulher de ambos os grupos descontinuou por amenorreia. CONCLUSãO: A taxa de continuação foi significativamente maior no grupo de anticoncepção durante os dos primeiros anos de uso. Amenorreia não foi motivo de descontinuação em ambos os grupos, sugerindo que a orientação a esse respeito foi adequada. Entretanto, a orientação referente ao longo período de sangramentos irregulares nos dois primeiros anos após a inserção, precisaria ser melhorado.

Phenotype and progression among patients with dilated cardiomyopathy and RBM20 mutations.

BACKGROUND: Over 70 genes that encode different cell components have been involved in the aetiology of dilated cardiomyopathy. Genotype-phenotype interactions are an unsolved problem, and to a large extent the effects of mutations in the expression mechanisms involved in the disease remain unknown, although associations are increasingly being established which have clinical and prognostic implications. METHODS AND RESULTS: The objective of our work was to describe our population that has cardiomyopathy associated with mutations in the gene RBM20, and study the genotype-phenotype relationship. We studied 8 cases undergoing follow-up at our Unit, and collected data for demographic, clinical and diagnostic testing variables. The mean age on diagnosis was 55 years [52-59], with a median follow-up of 31.5 months [26.0-67.3]. It is worth noting that 62.5% of the patients in our group had a history of cardiomyopathy in first degree relatives, and 37.5% of them had a family history of sudden death. One of the genetic variations of the sample was shared by three subjects who had no apparent family relationship with each other, and this variation had not been described in controls. It is also interesting that arrhythmic events were found in 37.5% of the sample, and 50% of patients had an indication for implantable cardiac defibrillator. CONCLUSION: This is the first analysis of patients with RBM20 mutations conducted in our country, and it indicates a profile with prominent arrhythmogenesis, a high penetrance of familial cardiomyopathy, and sudden death.

[SARS-CoV-2 seroprevalence among workers in a hospital in Madrid.] / Seroprevalencia frente a SARS-CoV-2 en los trabajadores de un hospital de Madrid.

OBJECTIVE: The impact of the first wave of the pandemic due to SARS-CoV-2 has been enormous in Spanish hospitals, being Madrid one of the most affected communities. The aim of this study was to analyze the impact of the pandemic on the workers of a hospital located in one of the most affected areas of Madrid (Alcalá de Henares) using a chemiluminescence immunoassay (CLIA). METHODS: We conducted a seroprevalence survey between June 25th and July 31st, 2020, over hospital workers from our institution. This study population was previously analyzed using a diagnostic approach which consisted of PCR and a serologic rapid test. For the present survey, prevalence of IgG antibodies against SARS-CoV-2 was studied using chemiluminescence. We analyzed the prevalence in healthcare workers (HCW) and non-healthcare workers (nHCW). Information about professional category, use of personal protective equipment (PPEs) and previous COVID-19 contacts was collected to determine the risk factors associated with SARS-CoV-2 seropositivity. Comparisons were made using the Mann-Whitney U test (continuous variables) and the two-tailed Fisher's exact test (categorical variables). Risk factors for seropositivity were analyzed using multivariate logistic regression. RESULTS: A total of 2,219 workers were included and 847 of them (38.2%) presented IgG antibodies against SARS-CoV-2, being seroprevalence significantly higher for HCW (39.3%) than for nHCW (30.7%, p=0.006). We found no differences regarding positivity rates compared with the first diagnostic approach for symptomatic and asymptomatic workers, or infected workers per professional category. In the multivariate logistic regression analysis, the use of PPE (protective, OR=0.65; 95%CI: 0.48-0.89; p<0.001) and previous contact with COVID-19 patients (risk factor, OR=1.93; 95%CI: 1.37-2.72; p<0.001) were independent factors that were associated with SARS-CoV-2 seropositivity. CONCLUSIONS: More than 38% of our workers presented IgG antibodies against SARS-CoV-2. We found no differences in the positivity rates between diagnostic approaches. The use of PPE (protective) and previous contact with COVID-19 patients (risk factor) were associated with infection.

OBJETIVO: El impacto de la primera oleada de la pandemia debida a SARS-CoV-2 ha sido enorme en los hospitales españoles, siendo Madrid una de las comunidades más afectadas. El objeto de este estudio fue analizar el impacto de la pandemia en los trabajadores de un hospital situado en una de las zonas más afectadas de Madrid (Alcalá de Henares) mediante un inmunoensayo de quimioluminiscencia. METODOS: Realizamos un estudio de seroprevalencia entre el 25 de junio y el 31 de julio de 2020 entre los trabajadores de nuestro centro, que se estudió previamente utilizando otra aproximación diagnóstica (PCR y test rápido serológico). Para este estudio, analizamos la prevalencia de anticuerpos tipo IgG contra SARS-CoV-2 mediante quimioluminiscencia. Analizamos la prevalencia en personal sanitario (PS) y no sanitario (PNS). Se recopiló información sobre la categoría profesional, uso de equipos de protección individual (EPI) y contactos COVID-19 para determinar los factores de riesgo asociados con la seropositividad. Las comparaciones se hicieron con el test de la U de Mann-Whitney (variables continuas) y la prueba exacta de Fisher de dos colas (variables categóricas). Los factores de riesgo para la seropositividad se analizaron mediante regresión logística multivariable. RESULTADOS: Incluimos a 2.219 trabajadores, de los cuales 847 (38,2%) presentaron anticuerpos IgG, siendo la seroprevalencia significativamente mayor en PS (39,3%) que en PNS (30,7%, p=0,006). No encontramos diferencias respecto a la primera aproximación diagnóstica en los porcentajes de positividad para personal sintomático, asintomático, o estratificando por categoría profesional. Mediante regresión logística multivariable, el uso de EPI (protector, OR=0,65; IC95%:0,48-0,89; p<0,001) y el contacto previo con pacientes COVID-19 (factor de riesgo, OR=1,93; IC95%:1,37-2,72; p<0,001) fueron factores asociados de forma independiente con la seropositividad. CONCLUSIONES: Más del 38% de nuestros trabajadores presentaron anticuerpos IgG contra SARS-CoV-2. No encontramos diferencias en los porcentajes de positividad entre ésta y la anterior aproximación diagnóstica. El uso de EPI (protector) y el contacto con pacientes con COVID-19 (factor de riesgo) se asociaron de forma independiente con la infección.

Cellular Origins of the Lymphatic Endothelium: Implications for Cancer Lymphangiogenesis.

The lymphatic system plays important roles in physiological and pathological conditions. During cancer progression in particular, lymphangiogenesis can exert both positive and negative effects. While the formation of tumor associated lymphatic vessels correlates with metastatic dissemination, increased severity and poor patient prognosis, the presence of functional lymphatics is regarded as beneficial for anti-tumor immunity and cancer immunotherapy delivery. Therefore, a profound understanding of the cellular origins of tumor lymphatics and the molecular mechanisms controlling their formation is required in order to improve current strategies to control malignant spread. Data accumulated over the last decades have led to a controversy regarding the cellular sources of tumor-associated lymphatic vessels and the putative contribution of non-endothelial cells to this process. Although it is widely accepted that lymphatic endothelial cells (LECs) arise mainly from pre-existing lymphatic vessels, additional contribution from bone marrow-derived cells, myeloid precursors and terminally differentiated macrophages, has also been claimed. Here, we review recent findings describing new origins of LECs during embryonic development and discuss their relevance to cancer lymphangiogenesis.

Comparison of In Vitro Efficacy of Six Disinfectants on the Hatching of Larval Eggs of Toxocara canis.

BACKGROUND: The environmental contamination with Toxocara canis eggs increases the risk of dissemination and transmission of the parasite in dogs and paratenic hosts such as humans. We aimed to evaluate different disinfectants to compare their effect on T. canis eggs. METHODS: For its realization, 850 embryonated eggs were obtained, which were suspended in a solution of 5% formaldehyde and distilled water in Eppendorf tubes. In the tubes containing the 850 embryonated eggs, researchers was added 0.5 mL of each solution (enzymatic solution, sodium hypochlorite, iodopovidone, quaternary of ammonium, benzalkonium chloride, and super oxidation solution). After mixing, an aliquot was taken, observed under the microscope, and the number of broken eggs counted at different times to find the most effective ovicidal moment. RESULTS: The enzymatic disinfectant present a significant difference (P = 0.05) with 276.06 broken eggs followed by ammonium with 105.20 broken eggs. After 10 min, the ammonium solution was the one that showed a significant difference of 50.50 hatched eggs, followed by the enzymatic 26.80 and hypochlorite 25.00 treatments. After 20 min, the enzymatic solution treatment showed a significant difference with the other solutions showing an increase of 98.80 broken eggs. In the 30 and 40-min times, only the enzymatic treatment showed a significant difference of 334.10 and 381.70 of broken eggs respectively. CONCLUSION: The enzymatic solution has the greatest ovicidal effect against the eggs of T. canis to present a greater number of broken eggs in a given time between 20 and 40 minutes.

Cocultures of human colorectal tumor spheroids with immune cells reveal the therapeutic potential of MICA/B and NKG2A targeting for cancer treatment.

BACKGROUND: Immunotherapies still fail to benefit colorectal cancer (CRC) patients. Relevant functional assays aimed at studying these failures and the efficacy of cancer immunotherapy in human are scarce. 3D tumor cultures, called tumor organoids or spheroids, represent interesting models to study cancer treatments and could help to challenge these issues. METHODS: We analyzed heterotypic cocultures of human colon tumor-derived spheroids with immune cells to assess the infiltration, activation and function of T and NK cells toward human colorectal tumors in vitro. RESULTS: We showed that allogeneic T and NK cells rapidly infiltrated cell line-derived spheroids, inducing immune-mediated tumor cell apoptosis and spheroid destruction. NKG2D, a key activator of cytotoxic responses, was engaged on infiltrating cells. We thus assessed the therapeutic potential of an antibody targeting the specific ligands of NKG2D, MICA and MICB, in this system. Anti-MICA/B enhanced immune-dependent destruction of tumor spheroid by driving an increased NK cells infiltration and activation. Interestingly, tumor cells reacted to immune infiltration by upregulating HLA-E, ligand of the inhibitory receptor NKG2A expressed by CD8 and NK cells. NKG2A was increased after anti-MICA/B treatment and, accordingly, combination of anti-MICA/B and anti-NKG2A was synergistic. These observations were ultimately confirmed in a clinical relevant model of coculture between CRC patients-derived spheroids and autologous tumor-infiltrating lymphocytes. CONCLUSIONS: Altogether, we show that tumor spheroids represent a relevant tool to study tumor-lymphocyte interactions on human tissues and revealed the antitumor potential of immunomodulatory antibodies targeting MICA/B and NKG2A.