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Metabolomics is a dynamic tool for elucidating biochemical changes in human health and disease. Metabolic profiles provide a close insight into physiological states and are highly volatile to genetic and environmental perturbations. Variation in metabolic profiles can inform mechanisms of pathology, providing potential biomarkers for diagnosis and assessment of the risk of contracting a disease. With the advancement of high-throughput technologies, large-scale metabolomics data sources have become abundant. As such, careful statistical analysis of intricate metabolomics data is essential for deriving relevant and robust results that can be deployed in real-life clinical settings. Multiple tools have been developed for both data analysis and interpretations. In this review, we survey statistical approaches and corresponding statistical tools that are available for discovery of biomarkers using metabolomics.
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Pesquisa Biomédica , Metabolômica , Humanos , Metabolômica/métodos , Metaboloma/genética , Biomarcadores/metabolismo , Análise de DadosRESUMO
Despite recent biomedical breakthroughs and large genomic studies growing momentum, the Middle Eastern population, home to over 400 million people, is underrepresented in the human genome variation databases. Here we describe insights from Phase 1 of the Qatar Genome Program with whole genome sequenced 6047 individuals from Qatar. We identified more than 88 million variants of which 24 million are novel and 23 million are singletons. Consistent with the high consanguinity and founder effects in the region, we found that several rare deleterious variants were more common in the Qatari population while others seem to provide protection against diseases and have shaped the genetic architecture of adaptive phenotypes. These results highlight the value of our data as a resource to advance genetic studies in the Arab and neighboring Middle Eastern populations and will significantly boost the current efforts to improve our understanding of global patterns of human variations, human history, and genetic contributions to health and diseases in diverse populations.
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Genoma Humano , Genômica , Consanguinidade , Genética Populacional , Genoma Humano/genética , Genômica/métodos , Humanos , Oriente Médio , Catar/epidemiologiaRESUMO
Several clinical studies indicated that the daily use of aspirin or acetylsalicylic acid reduces the cancer risk via cyclooxygenases (Cox-1 and Cox-2) inhibition. In addition, aspirin-induced Cox-dependent and -independent antitumor effects have also been described. Here we report, for the first time, that aspirin treatment of human glioblastoma cancer (GBM) stem cells, a small population responsible for tumor progression and recurrence, is associated with reduced cell proliferation and motility. Aspirin did not interfere with cell viability but induced cell-cycle arrest. Exogenous prostaglandin E2 significantly increased cell proliferation but did not abrogate the aspirin-mediated growth inhibition, suggesting a Cox-independent mechanism. These effects appear to be mediated by the increase of p21 waf1 and p27 Kip1 , associated with a reduction of Cyclin D1 and Rb1 protein phosphorylation, and involve the downregulation of key molecules responsible for tumor development, that is, Notch1, Sox2, Stat3, and Survivin. Our results support a possible role of aspirin as adjunctive therapy in the clinical management of GBM patients.
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In the central nervous system (CNS), oligodendrocytes are the glial element in charge of myelin formation. Obtaining an overall presence of oligodendrocyte precursor cells/oligodendrocytes (OPCs/OLs) in culture from different sources of NSCs is an important research area, because OPCs/OLs may provide a promising therapeutic strategy for diseases affecting myelination of axons. The present study was designed to differentiate human olfactory bulb NSCs (OBNSCs) into OPCs/OLs and using expression profiling (RT-qPCR) gene, immunocytochemistry, and specific protein expression to highlight molecular mechanism(s) underlying differentiation of human OBNSCs into OPCs/OLs. The differentiation of OBNSCs was characterized by a simultaneous appearance of neurons and glial cells. The differentiation medium, containing cAMP, PDGFA, T3, and all-trans-retinoic acid (ATRA), promotes OBNSCs to generate mostly oligodendrocytes (OLs) displaying morphological changes, and appearance of long cytoplasmic processes. OBNSCs showed, after 5 days in OLs differentiation medium, a considerable decrease in the number of nestin positive cells, which was associated with a concomitant increase of NG2 immunoreactive cells and few O4(+)-OPCs. In addition, a significant up regulation in gene and protein expression profile of stage specific cell markers for OPCs/OLs (CNPase, Galc, NG2, MOG, OLIG1, OLIG2, MBP), neurons, and astrocytes (MAP2, ß-TubulinIII, GFAP) and concomitant decrease of OBNSCs pluripotency markers (Oct4, Sox2, Nestin), was demonstrated following induction of OBNSCs differentiation. Taken together, the present study demonstrate the marked ability of a cocktail of factors containing PDGFA, T3, cAMP, and ATRA, to induce OBNSCs differentiation into OPCs/OLs and shed light on the key genes and pathological pathways involved in this process.
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AMP Cíclico/farmacologia , Células-Tronco Neurais/efeitos dos fármacos , Neurogênese/efeitos dos fármacos , Bulbo Olfatório/citologia , Oligodendroglia/efeitos dos fármacos , Fator de Crescimento Derivado de Plaquetas/farmacologia , Tretinoína/farmacologia , Tri-Iodotironina/farmacologia , Adulto , Biomarcadores/metabolismo , Forma Celular/efeitos dos fármacos , Células Cultivadas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Pessoa de Meia-Idade , Células-Tronco Neurais/metabolismo , Oligodendroglia/metabolismo , Fenótipo , Fatores de TempoRESUMO
Mesenchymal stem cells (MSCs) are multipotent cells that can differentiate into various cell types such as cartilage, bone, and fat cells. Recent studies have shown that induction of MSCs in vitro by growth factors including epidermal growth factor (EGF) and fibroblast growth factor (FGF2) causes them to differentiate into neural like cells. These cultures also express ChAT, a cholinergic marker; and TH, a dopaminergic marker for neural cells. To establish a protocol with maximum differentiation potential, we examined MSCs under three experimental culture conditions using neural induction media containing FGF2, EGF, BMP-9, retinoic acid, and heparin. Adipose-derived MSCs were extracted and expanded in vitro for 3 passages after reaching >80% confluency, for a total duration of 9 days. Cells were then characterized by flow cytometry for CD markers as CD44 positive and CD45 negative. MSCs were then treated with neural induction media and were characterized by morphological changes and Q-PCR. Differentiated MSCs expressed markers for immature and mature neurons; ß Tubulin III (TUBB3) and MAP2, respectively, showing the neural potential of these cells to differentiate into functional neurons. Improved protocols for MSCs induction will facilitate and ensure the reproducibility and standard production of MSCs for therapeutic applications in neurodegenerative diseases.
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Tecido Adiposo/citologia , Neurônios Colinérgicos/fisiologia , Neurônios Dopaminérgicos/fisiologia , Células-Tronco Mesenquimais/fisiologia , Células-Tronco Neurais/fisiologia , Neurogênese , Adulto , Linhagem da Célula , Separação Celular , Células Cultivadas , Neurônios Colinérgicos/efeitos dos fármacos , Neurônios Colinérgicos/metabolismo , Meios de Cultura/metabolismo , Neurônios Dopaminérgicos/efeitos dos fármacos , Neurônios Dopaminérgicos/metabolismo , Feminino , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Humanos , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/metabolismo , Proteínas Associadas aos Microtúbulos/metabolismo , Células-Tronco Neurais/efeitos dos fármacos , Células-Tronco Neurais/metabolismo , Neurogênese/efeitos dos fármacos , Fenótipo , Tubulina (Proteína)/metabolismoRESUMO
The Zebrafish has emerged to become a powerful vertebrate animal model for cardiovascular research in recent years. Its advantages include easy genetic manipulation, transparency, small size, low cost, and the ability to survive without active circulation at early stages of development. Sequencing the whole genome and identifying ortholog genes with human genome made it possible to induce clinically relevant cardiovascular defects via genetic approaches. Heart function and disturbed hemodynamics need to be assessed in a reliable manner for these disease models in order to reveal the mechanobiology of induced defects. This effort requires precise determination of blood flow patterns as well as hemodynamic stress (i.e., wall shear stress and pressure) levels within the developing heart. While traditional approach involves time-lapse brightfield microscopy to track cell and tissue movements, in more recent studies fast light-sheet fluorescent microscopes are utilized for that purpose. Integration of more complicated techniques like particle image velocimetry and computational fluid dynamics modeling for hemodynamic analysis holds a great promise to the advancement of the Zebrafish studies. Here, we discuss the latest developments in heart function and hemodynamic analysis for Zebrafish embryos and conclude with our future perspective on dynamic analysis of the Zebrafish cardiovascular system. Developmental Dynamics 246:868-880, 2017. © 2017 Wiley Periodicals, Inc.
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Coração/embriologia , Hemodinâmica/fisiologia , Peixe-Zebra/embriologia , Animais , Velocidade do Fluxo Sanguíneo , Embrião não Mamífero , Coração/fisiologia , Estresse Mecânico , Peixe-Zebra/fisiologiaRESUMO
Neural stem cells (NSCs) are multipotent self-renewing cells that could be used in cellular-based therapy for a wide variety of neurodegenerative diseases including Alzheimer's diseases (AD), Parkinson's disease (PD), amyotrophic lateral sclerosis (ALS), and multiple sclerosis (MS). Being multipotent in nature, they are practically capable of giving rise to major cell types of the nervous tissue including neurons, astrocytes, and oligodendrocytes. This is in marked contrast to neural progenitor cells which are committed to a specific lineage fate. In previous studies, we have demonstrated the ability of NSCs isolated from human olfactory bulb (OB) to survive, proliferate, differentiate, and restore cognitive and motor deficits associated with AD, and PD rat models, respectively. The use of carbon nanotubes (CNTs) to enhance the survivability and differentiation potential of NSCs following their in vivo engraftment have been recently suggested. Here, in order to assess the ability of CNTs to enhance the therapeutic potential of human OBNSCs for restoring cognitive deficits and neurodegenerative lesions, we co-engrafted CNTs and human OBNSCs in TMT-neurodegeneration rat model. The present study revealed that engrafted human OBNSCS-CNTs restored cognitive deficits, and neurodegenerative changes associated with TMT-induced rat neurodegeneration model. Moreover, the CNTs seemed to provide a support for engrafted OBNSCs, with increasing their tendency to differentiate into neurons rather than into glia cells. The present study indicate the marked ability of CNTs to enhance the therapeutic potential of human OBNSCs which qualify this novel therapeutic paradigm as a promising candidate for cell-based therapy of different neurodegenerative diseases.
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Nanomedicina/métodos , Nanotubos de Carbono , Degeneração Neural , Células-Tronco Neurais/transplante , Doenças Neurodegenerativas/cirurgia , Neurogênese , Neurônios/patologia , Bulbo Olfatório/citologia , Alicerces Teciduais , Compostos de Trialquitina , Animais , Comportamento Animal , Células Cultivadas , Cognição , Modelos Animais de Doenças , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Fluorescência Verde/genética , Humanos , Masculino , Aprendizagem em Labirinto , Microscopia de Fluorescência , Células-Tronco Neurais/metabolismo , Doenças Neurodegenerativas/induzido quimicamente , Doenças Neurodegenerativas/patologia , Doenças Neurodegenerativas/fisiopatologia , Neurônios/metabolismo , Fenótipo , Ratos Wistar , Tempo de Reação , Fatores de Tempo , TransfecçãoRESUMO
In this study, we aim to demonstrate the fate of allogenic adult human olfactory bulb neural stem/progenitor cells (OBNSC/NPCs) transplanted into the rat hippocampus treated with ibotenic acid (IBO), a neurotoxicant specific to hippocampal cholinergic neurons that are lost in Alzheimer's disease. We assessed their possible ability to survive, integrate, proliferate, and differentiate into different neuronal and glial elements: we also evaluate their possible therapeutic potential, and the mechanism(s) relevant to neuroprotection following their engraftment into the CNS milieu. OBNSC/NPCs were isolated from adult human olfactory bulb patients, genetically engineered to express GFP and human nerve growth factor (hNGF) by lentivirus-mediated infection, and stereotaxically transplanted into the hippocampus of IBO-treated animals and controls. Stereological analysis of engrafted OBNSCs eight weeks post transplantation revealed a 1.89 fold increase with respect to the initial cell population, indicating a marked ability for survival and proliferation. In addition, 54.71 ± 11.38%, 30.18 ± 6.00%, and 15.09 ± 5.38% of engrafted OBNSCs were identified by morphological criteria suggestive of mature neurons, oligodendrocytes and astrocytes respectively. Taken together, this work demonstrated that human OBNSCs expressing NGF ameliorate the cognitive deficiencies associated with IBO-induced lesions in AD model rats, and the improvement can probably be attributed primarily to neuronal and glial cell replacement as well as the trophic influence exerted by the secreted NGF.
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Doença de Alzheimer/terapia , Terapia Baseada em Transplante de Células e Tecidos , Fator de Crescimento Neural/biossíntese , Células-Tronco Neurais/transplante , Bulbo Olfatório/citologia , Animais , Astrócitos/metabolismo , Diferenciação Celular , Linhagem Celular , Proliferação de Células , Neurônios Colinérgicos/efeitos dos fármacos , Transtornos Cognitivos/terapia , Modelos Animais de Doenças , Proteínas de Fluorescência Verde/biossíntese , Proteínas de Fluorescência Verde/genética , Células HEK293 , Hipocampo/citologia , Humanos , Ácido Ibotênico/farmacologia , Masculino , Aprendizagem em Labirinto , Neovascularização Fisiológica , Fator de Crescimento Neural/genética , Células-Tronco Neurais/metabolismo , Oligodendroglia/metabolismo , Ratos , Ratos WistarRESUMO
Glucose-based peritoneal dialysis (PD) solutions dilate the parietal and visceral peritoneal microvasculature by endothelium-dependent mechanisms that primarily involve hyperosmolality. This PD-mediated dilation occurs by active intracellular glucose uptake and adenosine Al receptor activation, and by hyperosmolality-stimulated glibenclamide-sensitive potassium channels. Both pathways invoke NO as a second messenger for vasodilation. We hypothesized that during crystalloid-induced osmosis, the osmotic water flux through the transendothelial water-exclusive aquaporin 1 (AQP1) channels is the primary mechanism whereby the endothelium is being stimulated to instigate hyperosmolality-driven vasodilation. Four microvascular levels (diameters in the range 6 - 100 microm) were visualized by intravital videomicroscopy of the terminal ileum in anesthetized rats. Microvascular diameters and flow were measured after topical exposure to a 5% hypertonic mannitol or 2.5% glucose-based PD solution, at baseline and after brief tissue pre-treatment (with 0.1% glutaraldehyde for 10 seconds) or after combined tissue pre-treatment and pharmacologic blockade of AQP1 with HgCl2 (100 micromol/L). Vascular endothelial integrity was verified by the response to acetylcholine (10(-4) mol/L) and sodium nitroprusside (10(-4) mol/L). The hyperosmolar solutions both caused rapid and sustained vasodilation at all microvascular levels, which was not altered by tissue pre-treatment. Inhibition of AQP1 completely abolished the mannitol-induced vasodilation and markedly attenuated the PD fluid-mediated vasodilation. Neither glutaraldehyde pre-treatment nor HgCl2 affected tissue integrity or endothelial cell function. We conclude that the peritoneal microvascular vasodilation caused by hyperosmolar PD fluid is instigated by the osmotic water flux through AQP1. Clinical PD solutions have components other than hyperosmolality that can induce endothelium-dependent peritoneal microvascular vasodilation independent of the AQP1-mediated osmosis.
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Aquaporina 1/fisiologia , Permeabilidade Capilar/efeitos dos fármacos , Soluções para Diálise/farmacocinética , Glucose/farmacocinética , Peritônio/efeitos dos fármacos , Vasodilatação/efeitos dos fármacos , Animais , Aquaporina 1/efeitos dos fármacos , Soluções Cristaloides , Diuréticos Osmóticos/farmacologia , Endotélio Vascular/efeitos dos fármacos , Glutaral/farmacologia , Íleo/efeitos dos fármacos , Íleo/metabolismo , Soluções Isotônicas/farmacologia , Manitol/farmacologia , Cloreto de Mercúrio/farmacologia , Osmose , Diálise Peritoneal , Peritônio/irrigação sanguínea , Peritônio/metabolismo , RatosRESUMO
Peritoneal dialysis (PD) solutions dilate microvessels by undefined mechanisms. This vasodilation directly affects ultrafiltration and solute exchange during a PD dwell and is thought to account for the variable mass transfer area coefficient for small solutes during a glucose-based hypertonic dwell. We hypothesized that PD-mediated vasodilation occurs by endothelium-dependent mechanisms that involve endothelium energy-dependent K+ channels (K(ATP)), adenosine A1 receptor activation, and NO release. We used intravital videomicroscopy to study 3 levels of microvessels (A1 inflow arterioles about 100 microm diameter to pre-capillary A3 arterioles 10 - 15 microm diameter) in the terminal ileum of anesthetized rats under control conditions in vivo in a tissue bath. Ileum was bathed with hypertonic mannitol or 2.5% glucose-based PD solution (Delflex: Fresenius Medical Care North America, Waltham, MA, U.S.A.) with or without topical application of individual or combined specific inhibitors of the endothelium-dependent dilation pathways.: NO (L-NMMA), prostaglandin I2 (mefenamic acid), endothelium hyperpolarizing factor (glibenclamide), and adenosine A1 receptor antagonist (DPCPX). The mannitol and PD solutions induced rapid and sustained peritoneal vasodilation whose magnitude depended on microvascular level and osmotic solute. Combined inhibition of endothelium-dependent dilation pathways completely abolished the mannitol-induced hyperosmolality-mediated dilation at all microvascular levels, but selectively eliminated the PD solution-mediated A3 dilation. The K(ATP) and adenosine receptor antagonists, individually or combined, remarkably attenuated dilation in the smaller pre-capillary arterioles; NO inhibition, alone or combined with K(ATP) and adenosine receptor antagonists, eliminated the PD solution-induced dilation. The cyclooxygenase pathway is not involved in PD-induced dilation. Solutions for PD dilate the visceral peritoneal microvasculature by endothelium-dependent mechanisms, primarily the NO pathway. Adenosine receptor-activated NO release and K(ATP) channel-mediated endothelium hyperpolarization significantly contribute to vasodilation in the smaller peritoneal pre-capillary arterioles.
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Permeabilidade Capilar/efeitos dos fármacos , Soluções para Diálise/farmacocinética , Íleo/irrigação sanguínea , Diálise Peritoneal , Vasodilatação/efeitos dos fármacos , Antagonistas do Receptor A1 de Adenosina/farmacologia , Animais , Diuréticos Osmóticos/farmacocinética , Endotélio Vascular/efeitos dos fármacos , Inibidores Enzimáticos/farmacologia , Glucose/farmacocinética , Glibureto/farmacologia , Hipoglicemiantes/farmacologia , Íleo/efeitos dos fármacos , Manitol/farmacocinética , Ácido Mefenâmico/farmacologia , Peritônio/irrigação sanguínea , Peritônio/efeitos dos fármacos , Ratos , Xantinas/farmacologia , ômega-N-Metilarginina/farmacologiaRESUMO
BACKGROUND: Resting electrocardiogram (ECG) is a valuable non-invasive diagnostic tool used in clinical medicine to assess the electrical activity of the heart while the patient is resting. Abnormalities in ECG may be associated with clinical biomarkers and can predict early stages of diseases. In this study, we evaluated the association between ECG traits, clinical biomarkers, and diseases and developed risk scores to predict the risk of developing coronary artery disease (CAD) in the Qatar Biobank. METHODS: This study used 12-lead ECG data from 13,827 participants. The ECG traits used for association analysis were RR, PR, QRS, QTc, PW, and JT. Association analysis using regression models was conducted between ECG variables and serum electrolytes, sugars, lipids, blood pressure (BP), blood and inflammatory biomarkers, and diseases (e.g., type 2 diabetes, CAD, and stroke). ECG-based and clinical risk scores were developed, and their performance was assessed to predict CAD. Classical regression and machine-learning models were used for risk score development. RESULTS: Significant associations were observed with ECG traits. RR showed the largest number of associations: e.g., positive associations with bicarbonate, chloride, HDL-C, and monocytes, and negative associations with glucose, insulin, neutrophil, calcium, and risk of T2D. QRS was positively associated with phosphorus, bicarbonate, and risk of CAD. Elevated QTc was observed in CAD patients, whereas decreased QTc was correlated with decreased levels of calcium and potassium. Risk scores developed using regression models were outperformed by machine-learning models. The area under the receiver operating curve reached 0.84 using a machine-learning model that contains ECG traits, sugars, lipids, serum electrolytes, and cardiovascular disease risk factors. The odds ratio for the top decile of CAD risk score compared to the remaining deciles was 13.99. CONCLUSIONS: ECG abnormalities were associated with serum electrolytes, sugars, lipids, and blood and inflammatory biomarkers. These abnormalities were also observed in T2D and CAD patients. Risk scores showed great predictive performance in predicting CAD.
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Background: The current study explores the genetic underpinnings of cardiac arrhythmia phenotypes within Middle Eastern populations, which are under-represented in genomic medicine research. Methods: Whole-genome sequencing data from 14,259 individuals from the Qatar Biobank were used and contained 47.8% of Arab ancestry, 18.4% of South Asian ancestry, and 4.6% of African ancestry. The frequency of rare functional variants within a set of 410 candidate genes for cardiac arrhythmias was assessed. Polygenic risk score (PRS) performance for atrial fibrillation (AF) prediction was evaluated. Results: This study identified 1196 rare functional variants, including 162 previously linked to arrhythmia phenotypes, with varying frequencies across Arab, South Asian, and African ancestries. Of these, 137 variants met the pathogenic or likely pathogenic (P/LP) criteria according to ACMG guidelines. Of these, 91 were in ACMG actionable genes and were present in 1030 individuals (~7%). Ten P/LP variants showed significant associations with atrial fibrillation p < 2.4 × 10-10. Five out of ten existing PRSs were significantly associated with AF (e.g., PGS000727, p = 0.03, OR = 1.43 [1.03, 1.97]). Conclusions: Our study is the largest to study the genetic predisposition to arrhythmia phenotypes in the Middle East using whole-genome sequence data. It underscores the importance of including diverse populations in genomic investigations to elucidate the genetic landscape of cardiac arrhythmias and mitigate health disparities in genomic medicine.
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Pharmacogenetic (PGx)-informed medication prescription is a cutting-edge genomic application in contemporary medicine, offering the potential to overcome the conventional "trial-and-error" approach in drug prescription. The ability to use an individual's genetic profile to predict drug responses allows for personalized drug and dosage selection, thereby enhancing the safety and efficacy of treatments. However, despite significant scientific and clinical advancements in PGx, its integration into routine healthcare practices remains limited. To address this gap, the Qatar Genome Program (QGP) has embarked on an ambitious initiative known as QPGx-CARES (Qatar Pharmacogenetics Clinical Applications and Research Enhancement Strategies), which aims to set a roadmap for optimizing PGx research and clinical implementation on a national scale. The goal of QPGx-CARES initiative is to integrate PGx testing into clinical settings with the aim of improving patient health outcomes. In 2022, QGP initiated several implementation projects in various clinical settings. These projects aimed to evaluate the clinical utility of PGx testing, gather valuable insights into the effective dissemination of PGx data to healthcare professionals and patients, and identify the gaps and the challenges for wider adoption. QPGx-CARES strategy aimed to integrate evidence-based PGx findings into clinical practice, focusing on implementing PGx testing for cardiovascular medications, supported by robust scientific evidence. The current initiative sets a precedent for the nationwide implementation of precision medicine across diverse clinical domains.
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Farmacogenética , Medicina de Precisão , Humanos , Catar , Farmacogenética/métodos , Medicina de Precisão/métodos , Testes FarmacogenômicosRESUMO
Respiratory viruses in patients with chronic obstructive pulmonary disease (COPD) or asthma have not been characterised in Qatar. This study aimed to identify the most common viral strains responsible for respiratory tract infections in asthma/COPD patients (without exacerbations) in Qatar during the winter season (2008-2009). Nasal swabs from patients with asthma/COPD and respiratory symptoms were evaluated for 15 common viruses. 200 adult patients (190 with asthma and 10 with COPD) were enrolled. Viral infections were present in 36 out of 200 patients (18 %). Cough and wheezing were the most common symptoms. Rhinovirus was the most common causative agent, followed by coronaviruses. Our findings confirm previous reports of rhinovirus prevalence in respiratory tract infections in asthma/COPD. A countrywide survey to confirm our findings is warranted.
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Asma/virologia , Doença Pulmonar Obstrutiva Crônica/virologia , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Viroses/epidemiologia , Viroses/virologia , Adulto , Idoso , Asma/complicações , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Mucosa Nasal/virologia , Doença Pulmonar Obstrutiva Crônica/complicações , Catar/epidemiologia , Infecções Respiratórias/complicações , Estações do Ano , Viroses/complicações , Vírus/classificação , Vírus/isolamento & purificação , Adulto JovemRESUMO
Marine habitats are Earth's largest aquatic ecosystems, yet little is known about marine organism's genomes. Molecular studies can unravel their genetics print, thus shedding light on specie's adaptation and speciation with precise authentication. However, extracting high molecular weight DNA from marine organisms and subsequent DNA library preparation for whole genome sequencing is challenging. The challenges can be explained by excessive metabolites secretion that co-precipitates with DNA and barricades their sequencing. In this work, we sought to resolve this issue by describing an optimized isolation method and comparing its performance with the most commonly reported protocols or commercial kits: SDS/phenol-chloroform method, Qiagen Genomic Tips kit, Qiagen DNeasy Plant mini kit, a modified protocol of Qiagen DNeasy Plant kit, Qiagen DNeasy Blood and Tissue kit, and Qiagen Qiamp DNA Stool mini kit. Our method proved to work significantly better for different marine species regardless of their shape, consistency, and sample preservation, improving Oxford Nanopore Technologies sequencing yield by 39 folds for Spirobranchus sp. and enabling generation of almost 10 GB data per flow cell/run for Chrysaora sp. and Palaemon sp. samples.
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Host genomic information, specifically genomic variations, may characterize susceptibility to disease and identify people with a higher risk of harm, leading to better targeting of care and vaccination. Italy was the epicentre for the spread of COVID-19 in Europe, the first country to go into a national lockdown and has one of the highest COVID-19 associated mortality rates. Qatar, on the other hand has a very low mortality rate. In this study, we compared whole-genome sequencing data of 14398 adults and Qatari-national to 925 Italian individuals. We also included in the comparison whole-exome sequence data from 189 Italian laboratory-confirmed COVID-19 cases. We focused our study on a curated list of 3619 candidate genes involved in innate immunity and host-pathogen interaction. Two population-gene metric scores, the Delta Singleton-Cohort variant score (DSC) and Sum Singleton-Cohort variant score (SSC), were applied to estimate the presence of selective constraints in the Qatari population and in the Italian cohorts. Results based on DSC and SSC metrics demonstrated a different selective pressure on three genes (MUC5AC, ABCA7, FLNA) between Qatari and Italian populations. This study highlighted the genetic differences between Qatari and Italian populations and identified a subset of genes involved in innate immunity and host-pathogen interaction.
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COVID-19/genética , Predisposição Genética para Doença/genética , Interações entre Hospedeiro e Microrganismos/genética , Adulto , Alelos , COVID-19/epidemiologia , Controle de Doenças Transmissíveis , Suscetibilidade a Doenças/metabolismo , Exoma/genética , Feminino , Frequência do Gene/genética , Predisposição Genética para Doença/epidemiologia , Genética Populacional , Genômica/métodos , Humanos , Imunidade Inata/imunologia , Itália/epidemiologia , Masculino , Catar/epidemiologia , SARS-CoV-2/genética , SARS-CoV-2/patogenicidade , Sequenciamento do Exoma/métodos , Sequenciamento Completo do Genoma/métodosRESUMO
The chimeric antigen receptor T cell (CAR-T cell) immunotherapy currently represents a hot research trend and it is expected to revolutionize the field of cancer therapy. Promising outcomes have been achieved using CAR-T cell therapy for haematological malignancies. Despite encouraging results, several challenges still pose eminent hurdles before being fully recognized. Directing CAR-T cells to target a single tumour associated antigen (TAA) as the case in haematological malignancies might be much simpler than targeting the extensive inhibitory microenvironments associated with solid tumours. This review focuses on the basic principles involved in development of CAR-T cells, emphasizing the differences between humoral IgG, T-cell receptors, CAR and Fcγ-CR constructs. It also highlights the complex inhibitory network that is usually associated with solid tumours, and tackles recent advances in the clinical studies that have provided great hope for the future use of CAR-T cell immunotherapy. While current Fcγ-CR T cell immunotherapy is in pre-clinical stage, is expected to provide a sound therapeutic approach to add to existing classical chemo- and radio-therapeutic modalities.
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Imunoterapia Adotiva/métodos , Neoplasias/terapia , Receptores de Antígenos Quiméricos/administração & dosagem , Receptores de IgG/administração & dosagem , Animais , Humanos , Imunoterapia/métodos , Imunoterapia/tendências , Imunoterapia Adotiva/tendências , Neoplasias/imunologia , Receptores de Antígenos Quiméricos/imunologia , Receptores de IgG/imunologiaRESUMO
The global combat against MTB is limited by challenges in accurate affordable detection. In this study, a rapid, affordable, single tube system for detection of unamplified MTB16s rDNA was developed. Utilizing a AuNP based FRET system, this assay achieved a sensitivity and specificity of 98.6% and 90% respectively.
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DNA Bacteriano/isolamento & purificação , DNA Ribossômico/isolamento & purificação , Transferência Ressonante de Energia de Fluorescência/métodos , Nanopartículas Metálicas , Mycobacterium tuberculosis/isolamento & purificação , Tuberculose Pulmonar/diagnóstico , DNA Bacteriano/genética , DNA Ribossômico/genética , Transferência Ressonante de Energia de Fluorescência/economia , Ouro , Humanos , Mycobacterium tuberculosis/genética , RNA Ribossômico 16S/genética , Sensibilidade e Especificidade , Tuberculose Pulmonar/microbiologiaRESUMO
INTRODUCTION: Tuberculosis (TB) is a leading killer worldwide. End TB strategy aims at ending the TB epidemic by 2030. Early, accurate, and affordable diagnosis represents a cornerstone to achieve this goal. Innovative strategies for TB diagnostics have been introduced. However, the ideal assay is yet unavailable and conventional methods remain necessary for diagnosis. Unique properties of nanoparticles (NPs) have allowed their utilization in TB detection via targeting disease biomarkers. Area covered: Until now, around thirty-five TB NP-based assays have been partially or fully characterized. Accuracy, low-cost, and short time-to-result represent the common properties of proposed platforms. TB nanodiagnostics now encompass almost all clinical aspects of the disease including active TB, non-tuberculous mycobacteria, rifampicin resistant TB, TB/HIV co-infection, latent TB, and extra-pulmonary TB. This review summarizes state-of-the-art knowledge of TB nanodiagnostics for the last 10 years. Special consideration is given for fabrication concepts, detection strategies, and clinical performance using various clinical specimens. The potential of TB nanodiagnostics to fulfill the need for ideal MTB testing is assessed. Expert commentary: TB nanodiagnostics show promise to be ideal detection tools that can meet the rigorous demands to end the TB epidemic by 2030.
Assuntos
Nanopartículas/química , Tuberculose/diagnóstico , HumanosRESUMO
A platform for nucleic acid detection employing chitosan and chitosan coated gold nanoparticles (AuNPs) was developed. Mycobacterium tuberculosis (MTB) was used as a model target. MTB DNA was extracted from sputum using simple total nucleic acid extraction. Following amplification of MTB DNA, chitosan and AuNPs were added to samples. Free chitosan conjugated non-target DNA in negative samples, avoiding AuNP-DNA interaction and hence negative samples remained red. In positive samples, amplified DNA was capable of saturating free chitosan leading to AuNP aggregation where positive samples turned blue. Via visual color detection 15/16 MTB positive samples and 3/3 negative samples were correctly identified. This test is a 1-tube, 1-step assay reducing the risk of contamination in molecular laboratories and is a proof of concept on how chitosan; a cheap polymer could increase the sensitivity of AuNPs towards specific detection of nucleic acids without using target specific oligotargeters or expensive extraction kits.