Synthesis,Structural Analysis,and Biological Evaluation of Thioxoquinazoline Derivatives as Phosphodiesterase 7 Inhibitors

PDE7 inhibitors regulate pro‐inflammatory and immune T‐cell functions, and are a potentially novel class of drugs particularly useful for treatment of a wide variety of immune and inflammatory disorders. Structural optimization of thioxoquinazoline derivatives led to new compounds with very interesting profiles as PDE7 or PDE7/PDE4 dual inhibitors, which may be further developed as new drugs for inflammatory and neurological diseases.

     

MMP1 and MMP11 Expression in Peripheral Blood Mononuclear Cells upon Their Interaction with Breast Cancer Cells and Fibroblasts

Tumor-infiltrating immune cells phenotype is associated with tumor progression. However, little is known about the phenotype of the peripheral blood mononuclear cells (PBMC) from breast cancer patients. We investigated MMP1 and MMP11 expression in PBMC from breast cancer patients and we analyzed gene expression changes upon their interaction with cancer cells and cancer-associated fibroblasts (CAF). We measured the impact of PBMC on proinflammatory gene expression in breast cancer cells, normal fibroblast (NF), and CAF and the impact on proliferation and invasiveness capacity of breast cancer cells. Gene expression of MMP1 and MMP11 in PBMC from breast cancer patients (n = 54) and control (n = 28); expression of IL1A, IL6, IL17, IFNβ, and NFĸB in breast cancer cell lines (MCF-7 and MDA-MB-231); and, additionally, IL10 and MMP11 in CAF and NF were analyzed by qRT-PCR before and after co-culture. Our results show the existence of a subpopulation of breast cancer patients (25.9%) with very high levels of MMP11 gene expression in PBMC. Also, gene expression of MMP1 and MMP11 increases in PBMC after co-culture with breast cancer cell lines, NF or CAF. PBMC from healthy or breast cancer patients induce an increased proliferation rate on MCF-7 and an increased invasiveness capacity of MDA-MB-231. Finally, we show a differential expression profile of inflammatory genes in NF and CAF when co-cultured with control or breast cancer PBMC. We have observed that MMPs’ expression in PBMC is regulated by the microenvironment, while the expression of inflammatory genes in NF or CAF is differentially regulated by PBMC. These findings confirm the importance of the crosstalk between stromal cells and suggest that PBMC would play a role in promoting aggressive tumor behavior.     

Anti-Trypanosoma cruzi Activity of Metabolism Modifier Compounds

Chagas disease is caused by the protozoan parasite Trypanosoma cruzi and affects over 6 million people worldwide. Development of new drugs to treat this disease remains a priority since those currently available have variable efficacy and frequent adverse effects, especially during the long regimens required for treating the chronic stage of the disease. T. cruzi modulates the host cell-metabolism to accommodate the cell cytosol into a favorable growth environment and acquire nutrients for its multiplication. In this study we evaluated the specific anti-T. cruzi activity of nine bio-energetic modulator compounds. Notably, we identified that 17-DMAG, which targets the ATP-binding site of heat shock protein 90 (Hsp90), has a very high (sub-micromolar range) selective inhibition of the parasite growth. This inhibitory effect was also highly potent (IC₅₀ = 0.27 μmol L⁻¹) against the amastigote intracellular replicative stage of the parasite. Moreover, molecular docking results suggest that 17-DMAG may bind T. cruzi Hsp90 homologue Hsp83 with good affinity. Evaluation in a mouse model of chronic T. cruzi infection did not show parasite growth inhibition, highlighting the difficulties encountered when going from in vitro assays onto preclinical drug developmental stages.     

Early-Life Development of the Bifidobacterial Community in the Infant Gut

The establishment of the gut microbiota poses implications for short and long-term health. Bifidobacterium is an important taxon in early life, being one of the most abundant genera in the infant intestinal microbiota and carrying out key functions for maintaining host-homeostasis. Recent metagenomic studies have shown that different factors, such as gestational age, delivery mode, or feeding habits, affect the gut microbiota establishment at high phylogenetic levels. However, their impact on the specific bifidobacterial populations is not yet well understood. Here we studied the impact of these factors on the different Bifidobacterium species and subspecies at both the quantitative and qualitative levels. Fecal samples were taken from 85 neonates at 2, 10, 30, 90 days of life, and the relative proportions of the different bifidobacterial populations were assessed by 16S rRNA–23S rRNA internal transcribed spacer (ITS) region sequencing. Absolute levels of the main species were determined by q-PCR. Our results showed that the bifidobacterial population establishment is affected by gestational age, delivery mode, and infant feeding, as it is evidenced by qualitative and quantitative changes. These data underline the need for understanding the impact of perinatal factors on the gut microbiota also at low taxonomic levels, especially in the case of relevant microbial populations such as Bifidobacterium. The data obtained provide indications for the selection of the species best suited for the development of bifidobacteria-based products for different groups of neonates and will help to develop rational strategies for favoring a healthy early microbiota development when this process is challenged.     

Hierarchical structures of anodised cold gas sprayed titanium coatings

Cold gas spray (CGS) titanium coatings have been produced to obtain porous and rough coatings with enhanced mechanical performance. The coatings from optimal spraying conditions reached tensile strength values up to 40?MPa, shear strength up to 39?MPa and a loss mass of 37?mg/100 cycles in abrasive testing, values in accordance with the ASTM standards to be applied for orthopaedic joint prostheses. An innovative hierarchical structure (micro-nano) consisted of a TiO₂ nanotubes top layer obtained by anodisation onto a CGS Ti coating. The present paper focuses on the characterisation of both surfaces, as-sprayed CGS Ti layer and double Ti-TiO₂ layer, in terms of mechanical properties, surface topography and wettability (contact angle). There were not significant changes in micro-roughness, Ra～40?µm and Ra～30?µm, but a significant decrease in contact angle, from ≈26° up to 0°, was observed between these two structures. This behaviour indicates that the combination of the CGS?+?anodising results in promising high roughness superhydrophilic surfaces, ideal for biomedical applications.     

冷喷涂单颗粒铜在铝基体上的显微结构研究

采用冷喷涂法在铝(Al)基体上沉积单颗粒铜(Cu),利用聚焦离子束/电子束(FIB/SEM)系统精确定位并原位制备了完整单个颗粒Cu沉积在Al基体上的透射样品,分析其显微结构及形成原因。实验结果表明,撞击过程中温度与应力分布不均匀,导致沉积Cu颗粒不均匀形变。Cu/Al界面受影响较大:颗粒动能转化为形变能和热能,打破了界面处氧化膜,使界面附近温度迅速升高,发生动态再结晶,生成金属间化合物Cu_9Al_4;Cu颗粒内距界面越远的区域,受温度和应力的影响越小,其变形主要是通过晶体内位错增殖和移动;沉积颗粒顶部,远离Cu/Al界面,几乎不受应力和温度影响,保持原始显微结构。     

Antisense Oligonucleotide-Based Rescue of Aberrant Splicing Defects Caused by 15 Pathogenic Variants in ABCA4

The discovery of novel intronic variants in the ABCA4 locus has contributed significantly to solving the missing heritability in Stargardt disease (STGD1). The increasing number of variants affecting pre-mRNA splicing makes ABCA4 a suitable candidate for antisense oligonucleotide (AON)-based splicing modulation therapies. In this study, AON-based splicing modulation was assessed for 15 recently described intronic variants (three near-exon and 12 deep-intronic variants). In total, 26 AONs were designed and tested in vitro using a midigene-based splice system. Overall, partial or complete splicing correction was observed for two variants causing exon elongation and all variants causing pseudoexon inclusion. Together, our results confirm the high potential of AONs for the development of future RNA therapies to correct splicing defects causing STGD1.     

Virtual Screening of FDA-Approved Drugs against Triose Phosphate Isomerase from Entamoeba histolytica and Giardia lamblia Identifies Inhibitors of Their Trophozoite Growth Phase

Infectious diseases caused by intestinal protozoan, such as Entamoeba histolytica (E. histolytica) and Giardia lamblia (G. lamblia) are a worldwide public health issue. They affect more than 70 million people every year. They colonize intestines causing primarily diarrhea; nevertheless, these infections can lead to more serious complications. The treatment of choice, metronidazole, is in doubt due to adverse effects and resistance. Therefore, there is a need for new compounds against these parasites. In this work, a structure-based virtual screening of FDA-approved drugs was performed to identify compounds with antiprotozoal activity. The glycolytic enzyme triosephosphate isomerase, present in both E. histolytica and G. lamblia, was used as the drug target. The compounds with the best average docking score on both structures were selected for the in vitro evaluation. Three compounds, chlorhexidine, tolcapone, and imatinib, were capable of inhibit growth on G. lamblia trophozoites (0.05–4.935 μg/mL), while folic acid showed activity against E. histolytica (0.186 μg/mL) and G. lamblia (5.342 μg/mL).     

Urban data and urban design: A data mining approach to architecture education

The configuration of urban projects using Information and Communication Technologies is an essential aspect in the education of future architects. Students must know the technologies that will facilitate their academic and professional development, as well as anticipating the needs of the citizens and the requirements of their designs. In this paper, a data mining approach was used to outline the strategic requirements for an urban design project in an architecture course using a Project-Based Learning strategy. Informal data related to an award-winning public space (Gillett Square in London, UK) was retrieved from two social networks (Flickr and Twitter), and from its official website. The analysis focused on semantic, temporal and spatial patterns, aspects generally overlooked in traditional approaches. Text-mining techniques were used to relate semantic and temporal data, focusing on seasonal and weekly (work-leisure) cycles, and the geographic patterns were extracted both from geotagged pictures and by geocoding user locations. The results showed that it is possible to obtain and extract valuable data and information in order to determine the different uses and architectural requirements of an urban space, but such data and information can be challenging to retrieve, structure, analyze and visualize. The main goal of the paper is to outline a strategy and present a visualization of the results, in a way designed to be attractive and informative for both students and professionals – even without a technical background – so the conducted analysis may be reproducible in other urban data contexts.     

Mesostructured Thin Films as Responsive Optical Coatings of Photonic Crystals

A synthetic route is presented to attain high‐optical‐quality multilayered structures that result from coupling ordered mesoporous titanium oxide thin films to the surface of a dense one‐dimensional photonic crystal. Such architectures present spectrally well‐defined photon resonant modes localized in the outer coating that finely respond to physicochemically induced modifications of its pore volume. The potential of these porous coatings in detection of environmental changes through variations of the photonic response of the ensemble is demonstrated by performing isothermal optical reflectance measurements under controlled vapor‐pressure conditions.