Transplantation of hematopoietic stem cells (HSCs)—rare cells that can self‐renew and differentiate into cells of all hematopoietic lineages—is an important treatment option for patients with ...hematologic diseases. Key to the regenerative potential of HSCs is their natural in vivo microenvironment—the bone marrow niche—but in vitro engineering approaches to recapitulate the biochemical and biophysical properties of the bone marrow niche are still insufficiently explored. Herein, modular macroporous cryogels, mimicking the natural 3D architecture of trabecular bone, are seeded with human bone marrow‐derived mesenchymal stromal cells and decellularized to generate tissue‐specific extracellular matrix (ECM)‐decorated scaffolds. To recapitulate the physiological concentration of biomolecules in bone marrow, the cell culture medium is supplemented with macromolecular crowders that modulate the composition, ultrastructure, and mechanical properties of the cell‐deposited ECM. The ECM‐coated cryogel scaffolds generated under crowding conditions exhibit physicochemical properties similar to those of the endosteal niche of the bone marrow. The ECM‐functionalized cryogels presented could be used in the future for the ex vivo expansion of HSCs for transplantation and may also aid in the development of more realistic hematological disease models.
The development of a bone marrow niche model that closely mimics the natural 3D architecture of trabecular bone and the extracellular stromal compartment is reported. Macroporous cryogel scaffolds are functionalized with tissue‐specific, cell‐secreted ECM biopolymers. Macromolecular crowders enhance both ECM production and altered the microstructure of the secreted biopolymers.
Benznidazole, the primary drug used in Chagas’ disease treatment, has known side effects, which may limit its widespread use. Its low solubility could negatively interfere in the bioavailability, ...even accentuating the toxic effects. Cocrystals have been extensively used to modify and optimize physicochemical properties, but, as single-component raw materials, they are susceptible to the phenomenon of polymorphism. In this work, we report a trimorphic cocrystal containing a 1:1 ratio of benznidazole and salicylic acid. The crystalline structures of three polymorphs were elucidated by single-crystal X-ray diffraction. Moreover, several isostructural solvates were also synthesized and analyzed. The same carboxylic acid-imidazole supramolecular heterosynthon is present in the four forms, but the main structural feature is an extended column based on amide–amide hydrogen bonds. On the basis of the crystalline structures, the trimorphic system was classified as conformational and packing polymorphism. Furthermore, the dissolution profiles of the stable forms were determined and showed a significant solubility improvement over the raw material.
Background
Asthma is a heterogeneous disease with several phenotypes, endotypes and severity degrees, in which different T‐cell subpopulations are involved. These cells express specific miRNAs (i.e. ...inflamma‐miRs) that can be released to serum in exosomes after activation and be used as biomarkers of underlying inflammation. Thus, we aim to evaluate specific T‐cell miRNA signatures in serum exosomes from different subgroups of asthmatic patients.
Methods
Samples from healthy donors (N = 30) and patients (N = 119) with different asthma endotypes (T2high‐Atopic/T2high‐Non‐atopic/T2low) and severity degrees (mild/MA and moderate–severe/MSA) were used. Demographic, clinical, haematological and biochemical characteristics were collected. Twelve miRNAs previously associated with different Th subsets were preselected and their levels in serum exosome samples were measured using RTqPCR.
Results
We detected five miRNAs with high confidence in serum exosomes: miR‐16‐5p, miR‐21‐5p, miR‐126‐3p, miR146a‐5p and miR‐215‐5p. All of them, except miR‐16‐5p were upregulated in MSA patients compared to MA. A logistic regression model including each of these miRNAs was created to discriminate both conditions, rendering a ROC curve AUC of 0.896 (0.830–0.961). miR‐21‐5p and miR‐126‐3p, both involved in Th1/Th2 differentiation, were specifically augmented in T2high‐Atopic patients. Of note, all these changes were found in samples collected in autumn. On the contrary, IL‐6high patients with MSA, which were more obese, older, with higher neutrophil and basophil counts and TNF levels, displayed a decrease of miR‐21‐5p, miR‐126‐3p and miR‐146a‐5p.
Conclusion
Immune‐related miRNAs, including miR‐21‐5p, miR‐126‐3p, miR‐146a‐5p and miR‐215‐5p, can be used as clinically relevant non‐invasive biomarkers of the phenotype/endotype and severity of asthma.
We measured the levels of several inflamma‐miRs in serum exosomes from 30 healthy and 119 asthmatic donors with different phenotypes and severities. miR‐21‐5p, miR‐126‐3p, miR146a‐5p, and miR‐215‐5p increased in MSA vs. MA and a logistic regression model was fitted (AUC=0.896). miR‐21‐5p and miR‐126‐3p increased in T2high‐Atopic asthma, whereas miR‐21‐5p, miR‐126‐3p, and miR‐146a‐5p levels were downregulated in IL‐6high MSA patients.Abbreviations: AUC, area under the curve; MA, mild asthma; miRNA, microRNA; MSA, moderate–severe asthma
Abstract Huntington's disease (HD) patients and mouse models show learning and memory impairment even before the onset of motor symptoms. Deficits in hippocampal synaptic plasticity have been ...involved in the HD memory impairment. Several studies show that prostaglandin E2 (PGE2 ) EP2 receptor stimulates synaptic plasticity and memory formation. However, this role was not explored in neurodegenerative diseases. Here, we investigated the capacity of PGE2 EP2 receptor to promote synaptic plasticity and memory improvements in a model of HD, the R6/1 mice, by administration of the agonist misoprostol. We found that misoprostol increases dendritic branching in cultured hippocampal neurons in a brain-derived neurotrophic factor (BDNF)-dependent manner. Then, we implanted an osmotic mini-pump system to chronically administrate misoprostol to R6/1 mice from 14 to 18 weeks of age. We observed that misoprostol treatment ameliorates the R6/1 long-term memory deficits as analyzed by the T-maze spontaneous alternation task and the novel object recognition test. Importantly, administration of misoprostol promoted the expression of hippocampal BDNF. Moreover, the treatment with misoprostol in R6/1 mice blocked the reduction in the number of PSD-95 and VGluT-1 positive particles observed in hippocampus of vehicle-R6/1 mice. In addition, we observed an increase of cAMP levels in the dentate
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of WT and R6/1 mice treated with misoprostol. Accordingly, we showed a reduction in the number of mutant huntingtin nuclear inclusions in the dentate gyrus of R6/1 mice. Altogether, these results suggest a putative therapeutic effect of PGE2 EP2 receptor in reducing cognitive deficits in HD.
Preclinical data have demonstrated that BIBW 2992 is a potent irreversible inhibitor of ErbB1 (EGFR/HER1) and mutated ErbB1 receptors including the T790M variant, as well as ErbB2 (HER2). A phase I ...study of continuous once-daily oral BIBW 2992 was conducted to determine safety, maximum-tolerated dose, pharmacokinetics (PK), food effect, and preliminary antitumor efficacy.
Patients with advanced solid tumors were treated. PK evaluation was performed after the first dose and at steady-state.
Fifty-three patients received BIBW 2992 at 10 to 50 mg/d. BIBW 2992 was generally well-tolerated. The most common adverse effects included diarrhea, nausea, vomiting, rash, and fatigue. Dose-limiting toxicities included grade 3 rash (n = 2) and reversible dyspnea secondary to pneumonitis (n = 1). The recommended phase II dose was 50 mg/d. PK was dose proportional with a terminal elimination half-life ranging between 21.3 and 27.7 hours on day 1 and between 22.3 and 67.0 hours on day 27; BIBW 2992 exposure decreased after food intake. Three patients with non-small-cell lung carcinoma (NSCLC; two with in-frame exon 19 mutation deletions) experienced confirmed partial responses (PR) sustained for 24, 18, and 34 months, respectively. Two other patients (esophageal carcinoma and NSCLC) had nonconfirmed PRs. A patient with a PR at 10 mg/d progressed and developed symptomatic brain metastases, which subsequently regressed with an increased dose of 40 mg/d of BIBW 2992. A further seven patients had disease stabilization lasting > or = 6 months.
Continuous, daily, oral BIBW 2992 is safe and has durable antitumor activity. It is currently being evaluated in phase III trials.
Criminal minds create new psychoactive substances (NPS) to dribble the Drug Regulatory System. Early Warning Systems (EWA) were instituted worldwide to combat this practice. Brazilian government has ...started establishing EWA at a national level since August 2021. The role of drug analysis laboratories in EWA is very important and experts working at forensic laboratories are in the main position to detect NPS and any changes in the molecular structures. However, for this practice, it is necessary to use reliable analyses techniques for unequivocal identification. The goal of this work is to use single-crystal X-ray diffraction (XRD) to detect the presence of NPS and elucidate its molecular structure using single crystals from a seized blotter paper. XRD is also useful to prepare reference substances to produce entries to be included in Fourier transformed infrared spectroscopy (FTIR) and mass spectrometry databases, which are some of the most relevant routine techniques in forensic laboratories. This approach was used to study the NPS 25R-NBOH and 25I-NBOMe using XRD, FTIR, Raman, and nuclear magnetic resonance spectroscopies. The results showed that molecular structure methods must be used to elucidate NPS in an unambiguous manner and the potential of XRD use in the forensic area as a reference method.
Patellar tendinopathy is a common clinical problem, but its underlying pathophysiology remains poorly understood, primarily due to the absence of a representative experimental model. The most widely ...used method to generate such a model is collagenase injection, although this method possesses limitations. We developed an optimized rat model of patellar tendinopathy via the ultrasound-guided injection of collagenase mixed with a thermo-responsive Pluronic hydrogel into the patellar tendon of sixty male Wistar rats. All analyses were carried out at 3, 7, 14, 30, and 60 days post-injury. We confirmed that our rat model reproduced the pathophysiology observed in human patients through analyses of ultrasonography, histology, immunofluorescence, and biomechanical parameters. Tendons that were injured by the injection of the collagenase-Pluronic mixture exhibited a significant increase in the cross-sectional area (
< 0.01), a high degree of tissue disorganization and hypercellularity, significantly strong neovascularization (
< 0.01), important changes in the levels of types I and III collagen expression, and the organization and presence of intra-tendinous calcifications. Decreases in the maximum rupture force and stiffness were also observed. These results demonstrate that our model replicates the key features observed in human patellar tendinopathy. Collagenase is evenly distributed, as the Pluronic hydrogel prevents its leakage and thus, damage to surrounding tissues. Therefore, this model is valuable for testing new treatments for patellar tendinopathy.
Fragile X syndrome is the most common monogenetic cause of inherited intellectual disability and syndromic autism spectrum disorder. Fragile X syndrome is caused by an expansion (full mutation ≥200 ...CGGs repeats, normal 10-45 CGGs) of the fragile X mental retardation 1 (
) gene, epigenetic silencing of the gene, which leads to reduction or lack of the gene's product: the fragile X mental retardation protein. In this cross-sectional study, we assessed general and pharmacotherapy knowledge (GK and PTK) of fragile X syndrome and satisfaction with education in neurodevelopmental disorders (NDDs) among senior medical students in Serbia (N=348), Georgia (N=112), and Colombia (N=58). A self-administered 18-item questionnaire included GK (8/18) and PTK (7/18) components and self-assessment of the participants education in NDDs (3/18). Roughly 1 in 5 respondents had correct answers on half or more facts about fragile X syndrome (GK>PTK), which ranged similarly 5-7 in Serbia, 6-8 in Georgia, and 5-8 in Colombia, respectively. No cohort had an average value greater than 9 (60%) that would represent passing score "cut-off." None of the participants answered all the questions correctly. More than two-thirds of the participants concluded that they gained inadequate knowledge of NDDs during their studies, and that their education in this field should be more intense. In conclusion, there is a major gap in knowledge regarding fragile X syndrome among senior medical students in these three developing countries. The finding could at least in part be generalized to other developing countries aimed toward increasing knowledge and awareness of NDDs and fostering an institutional collaboration between developed and developing countries.
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