ABSTRACT
Next-generation RNA sequencing allows alternative splicing (AS) quantification with unprecedented resolution, with the relative inclusion of an alternative sequence in transcripts being commonly quantified by the proportion of reads supporting it as percent spliced-in (PSI). However, PSI values do not incorporate information about precision, proportional to the respective AS events' read coverage. Beta distributions are suitable to quantify inclusion levels of alternative sequences, using reads supporting their inclusion and exclusion as surrogates for the two distribution shape parameters. Each such beta distribution has the PSI as its mean value and is narrower when the read coverage is higher, facilitating the interpretability of its precision when plotted. We herein introduce a computational pipeline, based on beta distributions accurately modeling PSI values and their precision, to quantitatively and visually compare AS between groups of samples. Our methodology includes a differential splicing significance metric that compromises the magnitude of intergroup differences, the estimation uncertainty in individual samples, and the intragroup variability, being therefore suitable for multiple-group comparisons. To make our approach accessible and clear to both noncomputational and computational biologists, we developed betAS, an interactive web app and user-friendly R package for visual and intuitive differential splicing analysis from read count data.
Subject(s)
Alternative Splicing , Software , RNA Splicing , Sequence Analysis, RNA/methods , High-Throughput Nucleotide Sequencing/methodsABSTRACT
It is now established that many viruses that threaten public health establish condensates via phase transitions to complete their lifecycles, and knowledge on such processes may offer new strategies for antiviral therapy. In the case of influenza A virus (IAV), liquid condensates known as viral inclusions, concentrate the 8 distinct viral ribonucleoproteins (vRNPs) that form IAV genome and are viewed as sites dedicated to the assembly of the 8-partite genomic complex. Despite not being delimited by host membranes, IAV liquid inclusions accumulate host membranes inside as a result of vRNP binding to the recycling endocytic marker Rab11a, a driver of the biogenesis of these structures. We lack molecular understanding on how Rab11a-recycling endosomes condensate specifically near the endoplasmic reticulum (ER) exit sites upon IAV infection. We show here that liquid viral inclusions interact with the ER to fuse, divide, and slide. We uncover that, contrary to previous indications, the reported reduction in recycling endocytic activity is a regulated process rather than a competition for cellular resources involving a novel role for the host factor ATG9A. In infection, ATG9A mediates the removal of Rab11a-recycling endosomes carrying vRNPs from microtubules. We observe that the recycling endocytic usage of microtubules is rescued when ATG9A is depleted, which prevents condensation of Rab11a endosomes near the ER. The failure to produce viral inclusions accumulates vRNPs in the cytosol and reduces genome assembly and the release of infectious virions. We propose that the ER supports the dynamics of liquid IAV inclusions, with ATG9A facilitating their formation. This work advances our understanding on how epidemic and pandemic influenza genomes are formed. It also reveals the plasticity of recycling endosomes to undergo condensation in response to infection, disclosing new roles for ATG9A beyond its classical involvement in autophagy.
Subject(s)
Influenza A virus , Endoplasmic Reticulum/metabolism , Endosomes/metabolism , Influenza A virus/genetics , Microtubules/metabolism , Ribonucleoproteins/genetics , Ribonucleoproteins/metabolism , Autophagy-Related Proteins/genetics , Autophagy-Related Proteins/metabolism , Membrane Proteins/genetics , Membrane Proteins/metabolismABSTRACT
Gastric cancer is a dominating cause of cancer-associated mortality with limited therapeutic options. Here, we show that syndecan-4 (SDC4), a transmembrane proteoglycan, is highly expressed in intestinal subtype gastric tumors and that this signature associates with patient poor survival. Further, we mechanistically demonstrate that SDC4 is a master regulator of gastric cancer cell motility and invasion. We also find that SDC4 decorated with heparan sulfate is efficiently sorted in extracellular vesicles (EVs). Interestingly, SDC4 in EVs regulates gastric cancer cell-derived EV organ distribution, uptake, and functional effects in recipient cells. Specifically, we show that SDC4 knockout disrupts the tropism of EVs for the common gastric cancer metastatic sites. Our findings set the basis for the molecular implications of SDC4 expression in gastric cancer cells and provide broader perspectives on the development of therapeutic strategies targeting the glycan-EV axis to limit tumor progression.
Subject(s)
Stomach Neoplasms , Syndecan-4 , Humans , Heparitin Sulfate/metabolism , Neoplasm Invasiveness , Stomach Neoplasms/genetics , Syndecan-4/genetics , Syndecan-4/metabolismABSTRACT
BACKGROUND: The ileocecal valve (ICV) is considered to be one of the most difficult locations for endoscopic submucosal dissection (ESD). The objective of this study was to evaluate the efficacy and safety of traction-assisted ESD in this situation. METHODS: All patients who underwent traction-assisted ESD for an ICV lesion at three centers were identified from a prospective ESD database. En bloc and R0 rates were evaluated. Factors associated with non-R0 resection were explored. RESULTS: 106 patients with an ICV lesion were included. The median lesion size was 50 mm (interquartile range 38-60) and 58.5% (62/106) invaded the terminal ileum. The en bloc and R0 resection rates were 94.3% and 76.4%, respectively. Factors associated with non-R0 resection were lesions covering ≥75% of the ICV (odds ratio [OR] 0.21. 95%CI 0.06-0.76; P=0.02), and involving the anal lip (OR 0.36, 95%CI 0.13-0.99; P=0.04) or more than two sites on the ICV (OR 0.27, 95%CI 0.07-0.99; P=0.03). CONCLUSION: Traction-assisted ESD for treatment of ICV lesions was a safe and feasible option. Large lesions and anal lip involvement appeared to be factors predictive of difficulty.
ABSTRACT
BACKGROUND: This study evaluated the safety and efficacy of salvage endoscopic submucosal dissection (ESD) for Barrett's neoplasia recurrence after radiofrequency ablation (RFA). METHODS: Data from patients at 16 centers were collected for a multicenter retrospective study. Patients who underwent at least one RFA treatment for Barrett's esophagus and thereafter underwent further esophageal ESD for neoplasia recurrence were included. RESULTS: Data from 56 patients who underwent salvage ESD between April 2014 and November 2022 were collected. Immediate complications included one muscular tear (1.8%) treated with stent (Agree classification: grade IIIa). Two transmural perforations (3.6%; treated with clips) and five muscular tears (8.9%; two treated with clips) had no clinical impact and were not considered as adverse events. Seven patients (12.5%) developed strictures (grade IIIa), which were treated with balloon dilation. Histological analysis showed 36 adenocarcinoma, 17 high grade dysplasia, and 3 low grade dysplasia. En bloc and R0 resection rates were 89.3% and 66.1%, respectively. Resections were curative in 33 patients (58.9%), and noncurative in 22 patients (39.3%), including 11 "local risk" (19.6%) and 11 "high risk" (19.6%) resections. At the end of follow-up with a median time of 14 (0-75) months after salvage ESD, and with further endoscopic treatment if necessary (RFA, argon plasma coagulation, endoscopic mucosal resection, ESD), neoplasia remission ratio was 37/53 (69.8%) and the median remission time was 13 (1-75) months. CONCLUSION: In expert hands, salvage ESD was a safe and effective treatment for recurrence of Barrett's neoplasia after RFA treatment.
Subject(s)
Barrett Esophagus , Endoscopic Mucosal Resection , Esophageal Neoplasms , Neoplasm Recurrence, Local , Radiofrequency Ablation , Salvage Therapy , Humans , Barrett Esophagus/surgery , Barrett Esophagus/pathology , Endoscopic Mucosal Resection/adverse effects , Endoscopic Mucosal Resection/methods , Male , Female , Aged , Retrospective Studies , Salvage Therapy/methods , Middle Aged , Esophageal Neoplasms/surgery , Esophageal Neoplasms/pathology , Radiofrequency Ablation/adverse effects , Radiofrequency Ablation/methods , Adenocarcinoma/surgery , Adenocarcinoma/pathology , Esophageal Stenosis/etiology , Aged, 80 and over , Treatment Outcome , Esophagoscopy/methods , Esophagoscopy/adverse effectsABSTRACT
We investigate the physicochemical effects of pyroglutamination on the QHALTSV-NH2 peptide, a segment of cytosolic helix 8 of the human C-X-C chemokine G-protein-coupled receptor type 4 (CXCR4). This modification, resulting from the spontaneous conversion of glutamine to pyroglutamic acid, has significant impacts on the physicochemical features of peptides. Using a static approach, we compared the transformation in different conditions and experimentally found that the rate of product formation increases with temperature, underscoring the need for caution during laboratory experiments to prevent glutamine cyclization. Circular dichroism experiments revealed that the QHALTSV-NH2 segment plays a minor role in the structuration of H8 CXCR4; however, its pyroglutaminated analogue interacts differently with its chemical environment, showing increased susceptibility to solvent variations compared to the native form. The pyroglutaminated analogue exhibits altered behavior when interacting with lipid models, suggesting a significant impact on its interaction with cell membranes. A unique combination of atomic force microscopy and infrared nanospectroscopy revealed that pyroglutamination affects supramolecular self-assembly, leading to highly packed molecular arrangements and a crystalline structure. Moreover, the presence of pyroglumatic acid has been found to favor the formation of amyloidogenic aggregates. Our findings emphasize the importance of considering pyroglutamination in peptide synthesis and proteomics and its potential significance in amyloidosis.
Subject(s)
Amyloidosis , Glutamine , Humans , Peptides , Chemokines/chemistry , Cell Membrane/metabolism , Circular Dichroism , Receptors, CXCR4/metabolismABSTRACT
Previous studies have suggested that mammal life history varies along the fast-slow continuum and that, in eutherians, this continuum is linked to variation in the potential contribution of survival and reproduction to population growth rate (λ). Fast eutherians mature early, have large litters and short lifespans, and exhibit high potential contribution of age at first reproduction and fertility to λ, while slow eutherians show high potential contribution of survival to λ. However, marsupials have typically been overlooked in comparative tests of mammalian life-history evolution. Here, we tested whether the eutherian life-history pattern extends to marsupials, and show that marsupial life-history trade-offs are organized along two major axes: (i) the reproductive output and dispersion axis, and (ii) the fast-slow continuum, with an additional association between adult survival and body mass. Life-history traits that potentially drive changes in λ are similar in eutherians and marsupials with slow life histories, but differ in fast marsupials; age at first reproduction is the most important trait contributing to λ and fertility contributes little. Marsupials have slower life histories than eutherians, and differences between these clades may derive from their contrasting reproductive modes; marsupials have slower development, growth and metabolism than eutherians of equivalent size.
Subject(s)
Life History Traits , Marsupialia , Animals , Population Growth , Eutheria , FertilityABSTRACT
Cancer and normal cells use multiple antiapoptotic BCL2 proteins to prevent cell death. Therapeutic targeting of multiple BCL2 family proteins enhances tumor killing but is also associated with increased systemic toxicity. Here, we demonstrate that the dual targeting of MCL1 and BCL2 proteins using the small molecules S63845 and venetoclax induces durable remissions in mice that harbor human diffuse large B-cell lymphoma (DLBCL) tumors but is accompanied by hematologic toxicity and weight loss. To mitigate these toxicities, we encapsulated S63845 or venetoclax into nanoparticles that target P-selectin, which is enriched in tumor endothelial cells. In vivo and ex vivo imaging demonstrated preferential targeting of the nanoparticles to lymphoma tumors over vital organs. Mass spectrometry analyses after administration of nanoparticle drugs confirmed tumor enrichment of the drug while reducing plasma levels. Furthermore, nanoparticle encapsulation allowed 3.5- to 6.5-fold reduction in drug dose, induced sustained remissions, and minimized toxicity. Our results support the development of nanoparticles to deliver BH3 mimetic combinations in lymphoma and in general for toxic drugs in cancer therapy.
Subject(s)
Antineoplastic Agents/administration & dosage , Bridged Bicyclo Compounds, Heterocyclic/administration & dosage , Lymphoma, Large B-Cell, Diffuse/drug therapy , Myeloid Cell Leukemia Sequence 1 Protein/antagonists & inhibitors , Proto-Oncogene Proteins c-bcl-2/antagonists & inhibitors , Pyrimidines/administration & dosage , Sulfonamides/administration & dosage , Thiophenes/administration & dosage , Animals , Antineoplastic Agents/adverse effects , Antineoplastic Agents/therapeutic use , Bridged Bicyclo Compounds, Heterocyclic/adverse effects , Bridged Bicyclo Compounds, Heterocyclic/therapeutic use , Cell Line, Tumor , Drug Carriers/chemistry , Drug Delivery Systems , Female , Humans , Lymphoma, Large B-Cell, Diffuse/metabolism , Mice , Myeloid Cell Leukemia Sequence 1 Protein/metabolism , Nanoparticles/chemistry , Proto-Oncogene Proteins c-bcl-2/metabolism , Pyrimidines/adverse effects , Pyrimidines/therapeutic use , Sulfonamides/adverse effects , Sulfonamides/therapeutic use , Therapeutic Index , Thiophenes/adverse effects , Thiophenes/therapeutic useABSTRACT
BACKGROUND : During endoscopic submucosal dissection (ESD), the normal mucosa is cut under constant optical control. We studied whether a positive horizontal resection margin after a complete en bloc ESD predicts local recurrence. METHODS: In this European multicenter cohort study, patients with a complete en bloc colorectal ESD were selected from prospective registries. Cases were defined by a horizontal resection margin that was positive or indeterminate for dysplasia (HM1), whereas controls had a free resection margin (HM0). Low risk lesions with submucosal invasion (T1) and margins free of carcinoma were analyzed separately. The main outcome was local recurrence. RESULTS: From 928 consecutive ESDs (2011-2020), 354 patients (40â% female; mean age 67 years, median follow-up 23.6 months), with 308 noninvasive lesions and 46 T1 lesions, were included. The recurrence rate for noninvasive lesions was 1/212 (0.5â%; 95â%CI 0.02â%-2.6â%) for HM0 vs. 2/96 (2.1â%; 95â%CI 0.57â%-7.3â%) for HM1.âThe recurrence rate for T1 lesions was 1/38 (2.6â%; 95â%CI 0.14â%-13.5â%) for HM0 vs. 2/8 (25â%; 95â%CI 7.2â%-59.1â%) for HM1. CONCLUSION: A positive horizontal resection margin after an en bloc ESD for noninvasive lesions is associated with a marginal nonsignificant increase in the local recurrence rate, equal to an ESD with clear horizontal margins. This could not be confirmed for T1 lesions.
Subject(s)
Colorectal Neoplasms , Endoscopic Mucosal Resection , Humans , Female , Aged , Male , Margins of Excision , Prospective Studies , Cohort Studies , Colorectal Neoplasms/surgery , Colorectal Neoplasms/pathology , Treatment Outcome , Neoplasm Recurrence, Local/pathology , Retrospective StudiesABSTRACT
BACKGROUND : Endoscopic submucosal dissection (ESD) in colorectal lesions is technically demanding and a significant rate of noncurative procedures is expected. We aimed to assess the rate of residual lesions after a noncurative ESD for colorectal cancer (CRC) and to establish predictive scores to be applied in the clinical setting. METHODS : Retrospective multicenter analysis of consecutive colorectal ESDs. Patients with noncurative ESDs performed for the treatment of CRC lesions submitted to complementary surgery or with at least one follow-up endoscopy were included. RESULTS : From 2255 colorectal ESDs, 381 (17â%) were noncurative, and 135 of these were performed in CRC lesions. A residual lesion was observed in 24 patients (18â%). Surgery was performed in 96 patients and 76 (79â%) had no residual lesion in the colorectal wall or in the lymph nodes. The residual lesion rate for sm1 cancers was 0â%, and for >âsm1 cancers was also 0â% if no other risk factors were present. Independent risk factors for lymph node metastasis were poor differentiation and lymphatic permeation (NC-Lymph score). Risk factors for the presence of a residual lesion in the wall were piecemeal resection, poor differentiation, and positive/indeterminate vertical margin (NC-Wall score). CONCLUSIONS : Lymphatic permeation or poor differentiation warrant surgery owing to their high risk of lymph node metastasis, mainly in >âsm1 cancers. In the remaining cases, en bloc and R0 resections resulted in a low risk of residual lesions in the wall. Our scores can be a useful tool for the management of patients who undergo noncurative colorectal ESDs.
Subject(s)
Colorectal Neoplasms , Endoscopic Mucosal Resection , Humans , Endoscopic Mucosal Resection/adverse effects , Endoscopic Mucosal Resection/methods , Lymphatic Metastasis , Endoscopy , Retrospective Studies , Neoplasm, Residual , Colorectal Neoplasms/surgery , Colorectal Neoplasms/pathology , Treatment OutcomeABSTRACT
Doxorubicin (DOX) is a potent chemotherapeutic agent used against several cancer types. However, due to its cardiotoxic adverse effects, the use of this drug may be also life-threatening. Although most cancer patients are elderly, they are poorly represented and evaluated in pre-clinical and clinical studies. Considering this, the present work aims to evaluate inflammation and oxidative stress as the main mechanisms of DOX-induced cardiotoxicity, in an innovative approach using an experimental model constituted of elderly animals treated with a clinically relevant human cumulative dose of DOX. Elderly (18-20 months) CD-1 male mice received biweekly DOX administrations, for 3 weeks, to reach a cumulative dose of 9.0 mg/kg. One week (1W) or two months (2 M) after the last DOX administration, the heart was collected to determine both drug's short and longer cardiac adverse effects. The obtained results showed that DOX causes cardiac histological damage and fibrosis at both time points. In the 1W-DOX group, the number of nuclear factor kappa B (NF-κB) p65 immunopositive cells increased and a trend toward increased NF-κB p65 expression was seen. An increase of inducible nitric oxide synthase (iNOS) and interleukin (IL)-33 and a trend toward increased IL-6 and B-cell lymphoma-2-associated X (Bax) expression were seen after DOX. In the same group, a decrease in IL-1ß, p62, and microtubule-associated protein 1A/1B-light chain 3 (LC3)-I, p38 mitogen-activated protein kinase (MAPK) expression was observed. Contrariwise, the animals sacrificed 2 M after DOX showed a significant increase in glutathione peroxidase 1 and Bax expression with persistent cardiac damage and fibrosis, while carbonylated proteins, erythroid-2-related factor 2 (Nrf2), NF-κB p65, myeloperoxidase, LC3-I, and LC3-II expression decreased. In conclusion, our study demonstrated that in an elderly mouse population, DOX induces cardiac inflammation, autophagy, and apoptosis in the heart in the short term. When kept for a longer period, oxidative-stress-linked pathways remained altered, as well as autophagy markers and tissue damage after DOX treatment, emphasizing the need for continuous post-treatment cardiac monitoring.
Subject(s)
Antioxidants , Neoplasms , Animals , Male , Mice , Antioxidants/metabolism , Apoptosis , bcl-2-Associated X Protein/metabolism , Cardiotoxicity/etiology , Doxorubicin/pharmacology , Fibrosis , Inflammation/metabolism , NF-kappa B/metabolism , Oxidative Stress , Signal TransductionABSTRACT
BACKGROUND: The association between fetal growth restriction (FGR) and childhood neurodevelopmental delay is unclear and the evidence available to the present date shows conflicting results. Our aim was to analyse the impact of early-onset FGR on the neurodevelopmental outcome at 24 months of corrected age in very preterm infants. METHODS: Retrospective cohort study of very preterm infants (≤ 32 weeks' gestation) admitted to a neonatal intensive care unit between 1 January 2013-31 December 2019. The control group comprised appropriate for gestational age (AGA) newborns. Griffiths III Mental Development Scale was performed at 24 months of corrected age. RESULTS: 132 infants were included: 44 FGR and 88 AGA. Mean Global Development Quotient (GDQ) was lower for FGR infants (p = 0.004) even after adjusting for maternal and perinatal factors (ßadjusted -16.703; p = 0.009). The average scores for the neurodevelopmental domains were highest for personal-social-emotional skills (107.02 ± 16.34), followed by eye/hand coordination (105.61 ± 14.20) and foundation of learning skills (102.23 ± 13.74) and were lowest for gross motor (97.90 ± 11.88) and language/communication skills (96.39 ± 18.88). FGR had a significant negative impact on all domains except for gross motor skills. After adjustment, FGR continued to have a significant adverse impact on language/communication (ßadjusted -21.924; p = 0.013), eye/hand coordination (ßadjusted -15.446; p = 0.015) and foundation of learning skills (ßadjusted -15.211; p = 0.013). CONCLUSIONS: In very preterm infants, FGR was associated with a significantly increased risk of poor neurodevelopmental outcome at 24 months of corrected age compared to age-matched AGA infants.
Subject(s)
Fetal Growth Retardation , Infant, Premature, Diseases , Infant , Pregnancy , Female , Infant, Newborn , Humans , Child , Fetal Growth Retardation/etiology , Infant, Premature , Retrospective Studies , Infant, Very Low Birth Weight , Gestational AgeABSTRACT
Adeno-associated viral (AAV) vectors represent one of the leading platforms for gene delivery. Nevertheless, their small packaging capacity restricts their use for diseases requiring large-gene delivery. To overcome this, dual-AAV vector systems that rely on protein trans-splicing were developed, with the split-intein Npu DnaE among the most-used. However, the reconstitution efficiency of Npu DnaE is still insufficient, requiring higher vector doses. In this work, two split-inteins, Cfa and Gp41-1, with reportedly superior trans-splicing were evaluated in comparison with Npu DnaE by transient transfections and dual-AAV in vitro co-transductions. Both Cfa and Gp41-1 split-inteins enabled reconstitution rates that were over two-fold higher than Npu DnaE and 100% of protein reconstitution. The impact of different vector preparation qualities in split-intein performances was also evaluated in co-transduction assays. Higher-quality preparations increased split-inteins' performances by three-fold when compared to low-quality preparations (60-75% vs. 20-30% full particles, respectively). Low-quality vector preparations were observed to limit split-gene reconstitutions by inhibiting co-transduction. We show that combining superior split-inteins with higher-quality vector preparations allowed vector doses to be decreased while maintaining high trans-splicing rates. These results show the potential of more-efficient protein-trans-splicing strategies in dual-AAV vector co-transduction, allowing the extension of its use to the delivery of larger therapeutic genes.
Subject(s)
Protein Splicing , Trans-Splicing , Inteins , Gene Transfer Techniques , Drug PackagingABSTRACT
Traumatic spinal cord injury (SCI) initiates a cascade of cellular events, culminating in irreversible tissue loss and neuroinflammation. After the trauma, the blood vessels are destroyed. The blood-spinal cord barrier (BSCB), a physical barrier between the blood and spinal cord parenchyma, is disrupted, facilitating the infiltration of immune cells, and contributing to a toxic spinal microenvironment, affecting axonal regeneration. Understanding how the vascular constituents of the BSCB respond to injury is crucial to prevent BSCB impairment and to improve spinal cord repair. Here, we focus our attention on the vascular transcriptome at 3- and 7-days post-injury (dpi), during which BSCB is abnormally leaky, to identify potential molecular players that are injury-specific. Using the mouse contusion model, we identified Cd9 and Mylip genes as differentially expressed at 3 and 7 dpi. CD9 and MYLIP expression were injury-induced on vascular cells, endothelial cells and pericytes, at the injury epicentre at 7 dpi, with a spatial expression predominantly at the caudal region of the lesion. These results establish CD9 and MYLIP as two new potential players after SCI, and future studies targeting their expression might bring promising results for spinal cord repair.
Subject(s)
Endothelial Cells , Spinal Cord Injuries , Mice , Animals , Endothelial Cells/metabolism , Spinal Cord/metabolism , Spinal Cord Injuries/metabolism , Pericytes/metabolism , Disease Models, Animal , Gene Expression Profiling , Blood-Brain Barrier/metabolismABSTRACT
BACKGROUND: The presence of phenolic compounds in sunflower is well reported in the literature; however, knowledge is scarce when it comes to the composition of other secondary metabolites in this species and their by-products. This work evaluated, for the first time, the phytochemical composition of sunflower meal produced in Brazil. A combination of mixture design and central composite rotatable design 23 models was then applied to maximize the recovery of bioactive compounds using ecologically friendly solvents and concentrating by applying activated carbon, a sustainable adsorbent. The product of this extraction-concentration was also evaluated by an untargeted metabolomic approach using ultra-performance liquid chromatography coupled to mass spectrometry. RESULTS: A diverse and abundant profile of phenolic compounds was obtained from Brazilian sunflower meal: in total, 51 natural products were tentatively identified, 35 of which for the first time in sunflower. The sorption capacity of the activated charcoal, in the optimized process conditions, was effective in the separation and concentration of minority secondary metabolites. The ecofriendly extract proved to be enriched in plumberoside, p-coumaric acid, and alkaloids. CONCLUSIONS: Investigation of the phytochemical profile of sunflower meal produced in Brazil pointed to several secondary metabolites reported for the first time in sunflower samples, including phenolic compounds, alkaloids, and terpenes. The use of activated charcoal in an alkaline medium as an adsorbent for the concentration of these phytochemicals, from an aqueous extract, generated a potentially cost-effective, ecofriendly extract, enriched in minor metabolites, indicating a possible innovative way to selectively obtain these compounds from sunflower meal. © 2022 Society of Chemical Industry.
Subject(s)
Helianthus , Charcoal , Chromatography, High Pressure Liquid/methods , Phenols/analysis , Phytochemicals/chemistry , Plant Extracts/chemistryABSTRACT
A 71-year-old male was admitted on intensive care unit after endovascular aneurysm repair of ruptured infrarenal abdominal aortic aneurysm. 2 weeks later, he had multiple episodes of bloody diarrhea. Colonoscopy revealed diffuse dusky mucosal coloration with loss of vasculature pattern, diseased haustrations, and diffuse areas of pneumatosis, suggestive of severe colonic ischemia.
Subject(s)
Aortic Aneurysm, Abdominal , Blood Vessel Prosthesis Implantation , Colitis, Ischemic , Endovascular Procedures , Pneumatosis Cystoides Intestinalis , Male , Humans , Aged , Aortic Aneurysm, Abdominal/surgery , Colitis, Ischemic/diagnostic imaging , Colitis, Ischemic/surgery , Ischemia/diagnostic imaging , Ischemia/etiology , Pneumatosis Cystoides Intestinalis/diagnostic imagingABSTRACT
Common wheat (Triticum aestivum) is one of the most consumed staple foods used for bakery products. Outer layers of grain present a great diversity of bioactive compounds, especially phenolic compounds (PC). Free and bound PC were extracted from eight genotypes of whole wheat flours (WWF) presenting different technological classifications. These extracts were comprehensively characterized through untargeted metabolomics applying ultra-high-performance liquid chromatography-mass spectrometry (UHPLC-MSE) and spectrophotometric analyses. Chemical composition and colorimetry were also determined by classical analyses. Thirty-eight PC were tentatively identified by UHPLC-MSE belonging to three classes (phenolic acids, flavonoids, and other polyphenols), some of them identified in all WWF samples. Bound hydroxycinnamic acids were the main PC found in WWF, especially the trans-ferulic acid and its isomer. No difference was found in starch and protein contents, whereas low-quality flours showed a higher ash content than the superior and medium-quality flours. Total phenolic content (TPC) ranged between 124.5 and 171.4 mg GAE/100 g WWF, which bound PC were responsible for 60% of TPC. Omics data and multivariate statistical analyses were successfully applied to discern the phenolic profile of WWF from different genotypes and technological qualities. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-023-05665-8.
ABSTRACT
The NineTeen Complex (NTC), also known as pre-mRNA-processing factor 19 (Prp19) complex, regulates distinct spliceosome conformational changes necessary for splicing. During Drosophila midblastula transition, splicing is particularly sensitive to mutations in NTC-subunit Fandango, which suggests differential requirements of NTC during development. We show that NTC-subunit Salsa, the Drosophila ortholog of human RNA helicase Aquarius, is rate-limiting for splicing of a subset of small first introns during oogenesis, including the first intron of gurken Germline depletion of Salsa and splice site mutations within gurken first intron impair both adult female fertility and oocyte dorsal-ventral patterning, due to an abnormal expression of Gurken. Supporting causality, the fertility and dorsal-ventral patterning defects observed after Salsa depletion could be suppressed by the expression of a gurken construct without its first intron. Altogether, our results suggest that one of the key rate-limiting functions of Salsa during oogenesis is to ensure the correct expression and efficient splicing of the first intron of gurken mRNA. Retention of gurken first intron compromises the function of this gene most likely because it undermines the correct structure and function of the transcript 5'UTR.
Subject(s)
Body Patterning/genetics , Drosophila Proteins/metabolism , Drosophila melanogaster/physiology , Gene Expression Regulation, Developmental , Introns/genetics , RNA Splicing , Transforming Growth Factor alpha/metabolism , Animals , DNA-Binding Proteins/genetics , DNA-Binding Proteins/metabolism , Drosophila Proteins/genetics , Female , Infertility, Female/etiology , Infertility, Female/metabolism , Infertility, Female/pathology , Spliceosomes/genetics , Spliceosomes/metabolism , Transforming Growth Factor alpha/geneticsABSTRACT
Trisomy 18 is an autosomal chromosomal disorder characterized by the presence of an extra 18 chromosome. In the last decades, and as novel therapeutic options emerged, a paradigm shift on the treatments available to these children occurred, establishing the need to deepen the knowledge regarding the management/treatment of children diagnosed with trisomy 18. This retrospective cohort study sought to characterize the clinical path and survival of the children with the diagnosis of trisomy 18 followed in a tertiary pediatric hospital between 1995 and 2020. Medical records were reviewed, and epidemiological and clinical features and follow-up data were collected. Six patients were identified, two with mosaicism (33.3%) and four were female (66.7%). All had cardiovascular, cognitive, and physical development anomalies or minor congenital anomalies. Most presented neurological anomalies (n = 4, 66.7%) and feeding difficulties (n = 4, 66.7%). Four children (66.7%) required medical devices or equipment and all required chronic medication. Two children (33.3%) underwent surgical interventions. Four children (66.7%) were hospitalized in the last year of life. Three patients had a do not resuscitate order (50%) but only one child was referred to a pediatric palliative care team (16.7%). One-month, 1-year, and 10-year survival were 66.7% (n = 4), 33.3% (n = 2, both with mosaicism), and 16.7% (n = 1, with mosaicism) respectively. CONCLUSIONS: Knowledge of the multiple comorbidities and complex care needs of children with this syndrome is crucial. Every-day care and decisions about invasive treatments may raise ethical issues. Early referral to pediatric palliative care teams is essential to promote a holistic advanced care plan for both the patient and his family. WHAT IS KNOWN: ⢠The increase in survival and the high morbimortality that trisomy 18 still entails demands a careful deliberation on the use of invasive treatment. WHAT IS NEW: ⢠Recent studies show that the labels of "incompatible with life"/"lethal" are not adequate, establishing a need to change this mindset. ⢠The development of pediatric palliative care teams in the last decade and early referral allow for an optimal individualized advanced care plan. Under-referral to pediatric palliative care teams persists and efforts must be made to increase awareness of their existence and role in patient care.