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Three-dimensional (3D) culture provides a biomimicry of the naive microenvironment that can support cell proliferation, differentiation, and regeneration. Some growth factors, such as epidermal growth factor (EGF), facilitate normal meiosis during oocyte maturation in vivo. In this study, a scaffold-based 3D coculture system using purified alginate was applied to induce oocyte differentiation from mouse embryonic stem cells (mESCs). mESCs were induced to differentiate into oocyte-like cells using embryoid body protocol in the two-dimensional or 3D microenvironment in vitro. To increase the efficiency of the oocyte-like cell differentiation from mESCs, we employed a coculture system using ovarian granulosa cells in the presence or absence of epidermal growth factor (+EGF or -EGF) for 14 days and then the cells were assessed for germ cell differentiation, meiotic progression, and oocyte maturation markers. The cultures exposed to EGF in the alginate-based 3D microenvironment showed the highest level of premeiotic (Oct4 and Mvh), meiotic (Scp1, Scp3, Stra8, and Rec8), and oocyte maturation (Gdf9, Cx37, and Zp2) marker genes (p less then .05) in comparison to other groups. According to the gene-expression patterns, we can conclude that alginate-based 3D coculture system provided a highly efficient protocol for oocyte-like cell differentiation from mESCs. The data showed that this culture system along with EGF improved the rate of in vitro oocyte-like cell differentiation.Numerous drug-drug interaction (DDI) trials have to be conducted in healthy volunteers based on current regulatory guidelines. Because the worst-case scenario of strong cytochrome P450 (CYP) inhibitors has to be tested, the results and their validity have to be balanced with the risk to volunteer safety. The use of ketoconazole in clinical DDI studies has been discouraged by regulatory agencies due to an alleged risk of liver injury. In order to reduce the risk to healthy volunteers, we carried out a study with single-day exposure to each of 6 perpetrator azole fungistatic drugs. They were evaluated regarding their CYP3A inhibition using microdosed midazolam and a limited sampling strategy. Ratios of areas under the concentration-time curves ranged from 1.93 with isavuconazole to 8.42 with ketoconazole. The highest number of adverse events occurred with voriconazole, followed by ketoconazole; 2 dropouts occurred due to adverse events following itraconazole administration. Literature data on adverse events of azole fungistatic drugs in DDI trials are rare and inconclusive. Only in recent years with the newer drugs are they more precise and reliable. It can be concluded that the duration of preexposure of perpetrator drugs can be reduced to 1 hour before administration of the victim drug. This still can be sufficient to achieve the scientific objectives of the trial with the lowest possible risk.On November 5th 2015, Samarco iron ore mining operations released approximately 50 million m3 of mining waste into the environment, due to a dam collapse. Aiming at understanding the potential effects on the Doce River, different regions of the course of tailings were monitored using acute and chronic ecotoxicological tests with four species Ceriodaphnia dubia, Daphnia similis, Danio rerio and Vibrio fischeri. The results showed no acute toxicity for water column organisms. However, chronic toxicity were observed with the tests with the microcrustacean C. dubia, mainly related to the physical effects of the passage of flood wave and increased suspended solids, since toxicity was reduced after filtering the samples. The results showed different magnitude impact among upper, middle and lower Doce River, with greater impact close to the dam failure area. This article is protected by copyright. All rights reserved.Background Four commercial porcine reproductive and respiratory syndrome virus (PRRSV) modified-live vaccines (MLV) was compared to protect growing pigs against dual challenge of PRRSV-1 and PRRSV-2. Methods Two of the vaccines were based on PRRSV-1, and two on PRRSV-2. A total of 72 PRRSV-naïve pigs were divided into six groups (12 pigs/group). Results Two PRRSV-1 MLV-vaccinated and two PRRSV-2 MLV-vaccinated groups reduced significantly (p less then .05) genomic copies of PRRSV-1 in their sera compared to the unvaccinated challenged group. Two PRRSV-2 MLV-vaccinated groups reduced significantly (p less then .05) fewer genomic copies of PRRSV-2 in their sera whereas two PRRSV-1 MLV-vaccinated groups were unable to reduce genomic copies of PRRSV-2 compared to unvaccinated challenged groups. Two PRRSV-1 MLV-vaccinated groups induced a stronger PRRSV-1 specific IFN-γ-SC response, while two PRRSV-2 MLV-vaccinated groups induced a stronger PRRSV-2 specific IFN-γ-SC response. Two PRRSV-2 MLV-vaccinated groups showed significantly (p less then .05) lower mean macroscopic and microscopic lung lesion scores compared to two PRRSV-1 MLV-vaccinated groups. Conclusions These data demonstrated that two PRRSV-2 vaccines were efficacious and exhibited similar protection while, two PRRSV-1 vaccines were largely ineffective against the dual challenge.The ability of the bacterial pathogen Mycobacterium tuberculosis to adapt and survive within human cells to disseminate to other individuals and cause active disease is poorly understood. Research supports that as M. tuberculosis adapts to stressors encountered in the host, it exhibits variable physiological and metabolic states that are time and niche-dependent. Challenges associated with effective treatment and eradication of tuberculosis (TB) are in part attributed to our lack of understanding of these different mycobacterial phenotypes. This is mainly due to a lack of suitable tools to effectively identify/detect heterogeneous bacterial populations, which may include small, difficult-to-culture subpopulations. find more Importantly, flow cytometry allows rapid and affordable multiparametric measurements of physical and chemical characteristics of single cells, without the need to preculture cells. Here, we summarize current knowledge of flow cytometry applications that have advanced our understanding of the physiology of M.