Radiolabeling of stem cells with a positron emitting radioisotope represents a major advancement in regenerative biotherapy enabling non-invasive imaging. To assess the value of such an approach in a ...clinically relevant scenario, the tolerability and therapeutic aptitude of 89Zrzirconium-p-isothiocyanatobenzyl-desferrioxamine (89ZrZr-DBN) labeled human cardiopoietic stem cells (CPs) were evaluated in a model of ischemic heart failure.INTRODUCTIONRadiolabeling of stem cells with a positron emitting radioisotope represents a major advancement in regenerative biotherapy enabling non-invasive imaging. To assess the value of such an approach in a clinically relevant scenario, the tolerability and therapeutic aptitude of 89Zrzirconium-p-isothiocyanatobenzyl-desferrioxamine (89ZrZr-DBN) labeled human cardiopoietic stem cells (CPs) were evaluated in a model of ischemic heart failure.89ZrZr-DBN based radiolabeling of human CPs yielded 89ZrZr-DBN-CPs with radioactivity yield of 0.70 ± 0.20 MBq/106 cells and excellent label stability. Compared to unlabeled cell counterparts, 89ZrZr-DBN-CPs maintained morphology, viability, and proliferation capacity with characteristic expression of mesodermal and pro-cardiogenic transcription factors defining the cardiopoietic phenotype. Administered in chronically infarcted murine hearts, 89ZrZr-DBN-CPs salvaged cardiac pump failure, documented by improved left ventricular ejection fraction not inferior to unlabeled CPs and notably superior to infarcted hearts without cell treatment.METHODS AND RESULTS89ZrZr-DBN based radiolabeling of human CPs yielded 89ZrZr-DBN-CPs with radioactivity yield of 0.70 ± 0.20 MBq/106 cells and excellent label stability. Compared to unlabeled cell counterparts, 89ZrZr-DBN-CPs maintained morphology, viability, and proliferation capacity with characteristic expression of mesodermal and pro-cardiogenic transcription factors defining the cardiopoietic phenotype. Administered in chronically infarcted murine hearts, 89ZrZr-DBN-CPs salvaged cardiac pump failure, documented by improved left ventricular ejection fraction not inferior to unlabeled CPs and notably superior to infarcted hearts without cell treatment.The present study establishes that 89ZrZr-DBN labeling does not compromise stem cell identity or efficacy in the setting of heart failure, offering a non-invasive molecular imaging platform to monitor regenerative biotherapeutics post-transplantation.CONCLUSIONThe present study establishes that 89ZrZr-DBN labeling does not compromise stem cell identity or efficacy in the setting of heart failure, offering a non-invasive molecular imaging platform to monitor regenerative biotherapeutics post-transplantation.
Ectopic expression of pluripotency gene sets provokes nuclear reprogramming in permissive somatic tissue environments, generating nduced pluripotent stem (iPS) cells. The evolutionary conserved ...function of sternness orthologs was tested here through interspecies transduction. A spectrum of human immunodeficiency virus (HIV)‐based lentiviral vectors was designed, and point mutations in the HIV‐1 capsid region were identified for efficient infectivity and expanded transspecies tropism. Human pluripotent gene sequences, OCT3/4, SOX2, KLF4, and c‐MYC, packaged into engineered lentiviral expression vectors achieved consistent expression in nonhuman fibroblasts. Despite variation in primary amino acid sequence between species, introduction of human pluripotent genes produced cell lines with embryonic stem cell‐like morphology. Transduced fibroblasts differentiated in vitro into all three germ layers according to gastrulation gene expression profiles, and formed in vivo teratoma with multilineage potential. Reprogrammed progeny incorporated into nonhuman morula to produce blastomeres capable of developing into chimeric embryos with competent organogenesis. This model system establishes a prototypic approach to examine consequences of human sternness factors‐induced reprogramming in the context of normal embryonic development by exploiting nonhuman early‐stage embryos. Thus, ectopic xenotransduction across species unmasks the promiscuous nature of sternness induction, suggesting evolutionary selection of core processes for somatic tissue reprogramming.
1 Division of Cardiovascular Diseases, Departments
of Medicine and Molecular Pharmacology and Experimental
Therapeutics, and 2 Department of Biochemistry and
Molecular Biology, Mayo Clinic, Mayo ...Foundation, Rochester,
Minnesota 55905; and 3 Center for Molecular Life
Sciences, University Medical Center, University of Nijmegen, Nijmegen
6500, The Netherlands
Deletion of the major
adenylate kinase AK1 isoform, which catalyzes adenine nucleotide
exchange, disrupts cellular energetic economy and compromises metabolic
signal transduction. However, the consequences of deleting the
AK1 gene on cardiac energetic dynamics and performance in
the setting of ischemia-reperfusion have not been determined.
Here, at the onset of ischemia, AK1 knockout mice hearts
displayed accelerated loss of contractile force compared with wild-type
controls, indicating reduced tolerance to ischemic stress. On
reperfusion, AK1 knockout hearts demonstrated reduced nucleotide
salvage, resulting in lower ATP, GTP, ADP, and GDP levels and an
altered metabolic steady state associated with diminished
ATP-to-P i and creatine phosphate-to-P i ratios. Postischemic AK1 knockout hearts maintained ~40% of
-phosphoryl turnover, suggesting increased phosphotransfer flux
through remaining adenylate kinase isoforms. This was associated with
sustained creatine kinase flux and elevated cellular
glucose-6-phosphate levels as the cellular energetic system adapted to
deletion of AK1. Such metabolic rearrangements, along with sustained
ATP-to-ADP ratio and total ATP turnover rate, maintained
postischemic contractile recovery of AK1 knockout hearts at
wild-type levels. Thus deletion of the AK1 gene reveals that
adenylate kinase phosphotransfer supports myocardial function on
initiation of ischemic stress and safeguards intracellular
nucleotide pools in postischemic recovery.
energy metabolism; adenine nucleotides; glycolysis; phosphotransfer; oxygen-18 phosphoryl labeling; phosphorus-31 nuclear
magnetic resonance
The burden of disease and injury in Serbia Janković, Slavenka; Vlajinac, Hristina; Bjegović, Vesna ...
European journal of public health,
02/2007, Letnik:
17, Številka:
1
Journal Article
Recenzirano
Odprti dostop
Background: In the last decade of the 20th century, a considerable effort has been put into the development of summary measures of population health that combine information on mortality and ...non-fatal health outcomes. We used the DALYs (Disability adjusted life years) method to assess the burden of disease and injury in the population of Serbia. Methods: Our study, largely based on the methods developed for the Global burden of disease study, was conducted between October 2002 and September 2003. DALYs, stratified by gender and age, were calculated for 18 selected health conditions for the population of Serbia, Serbia and Montenegro for 2000. Years of life lost (YLL) were calculated using country mortality statistics, while years lived with disability (YLD) were calculated using different sources of information. Also, the YLD/YYL ratio and age-adjusted rates of DALYs were calculated. Results: Ischaemic heart disease, cerebrovascular diseases, lung cancer, unipolar depressive disorders, and diabetes mellitus were responsible for almost two-thirds (70%) of the total burden of 18 selected disorders in Serbia 2000. The leading five causes for males were ischaemic heart disease (26.1 DALY per 1000), stroke (17.9), lung cancer (12.7), road traffic accidents (6.5), and self-inflicted injuries (5.5). For females, the leading five causes were stroke (18.1 DALY per 1000), ischaemic heart disease (14.1), depression (8.7), breast cancer (6.1), and diabetes mellitus (5.2). Conclusions: The final results of the study have shown that the national health priority areas should cover cardiovascular diseases, cancers, and mental health.
The study of hemodynamic alterations following the creation of an arteriovenous fistula (AVF) is relevant to vascular adaptive responses and hemodialysis access dysfunction. This study examined such ...alterations in a murine AVF created by anastomosing the carotid artery to the jugular vein. AVF blood flow was markedly increased due to reduced AVF vascular resistance. Despite such markedly increased basal blood flow, AVF blood flow further increased in response to acetylcholine. This AVF model exhibited increased cardiac output and decreased systemic vascular resistance; the kidney, in contrast, exhibited decreased blood flow and increased vascular resistance. Augmentation in AVF blood flow was attended by increased arterial heme oxygenase-1 (HO-1) mRNA and protein expression, the latter localized to smooth muscle cells of the AVF artery; AVF blood flow was substantially reduced in HO-1(-/-) mice compared with HO-1(+/+) mice. Finally, in a murine model of a representative disease known to exhibit impaired hemodynamic responses (sickle cell disease), the creation of an AVF was attended by decreased AVF flow and impaired AVF function. We conclude that this AVF model exhibits markedly increased AVF blood flow, a vasodilatory reserve capacity, increased cardiac output, decreased renal blood flow, and a dependency on intact hemodynamic responses, in general, and HO-1 expression, in particular, in achieving and maintaining AVF blood flow. We suggest that these findings support the utility of this model in investigating the basis for and the consequences of hemodynamic stress, including shear stress, and the pathobiology of hemodialysis AVF dysfunction.
In this paper, a genetic algorithm-based optimization is used to simultaneously minimize two competing objectives guiding the operation of the Jefferson Lab’s Continuous Electron Beam Accelerator ...Facility linacs: cavity heat load and radio frequency cavity trip rates. The results represent a significant improvement to the standard linac energy management tool and thereby could lead to a more efficient Continuous Electron Beam Accelerator Facility configuration. This study also serves as a proof of principle of how a genetic algorithm can be used for optimizing other linac-based machines.
Embryonic stem cell differentiation recapitulates the diverse phenotypes of a developing embryo, traceable according to markers of lineage specification. At gastrulation, the vascular endothelial ...growth factor (VEGF) receptor, Flk-1 (KDR), identifies a mesoderm-restricted potential of embryonic stem cells. The multi-lineage propensity of Flk-1
+ progenitors mandates the mapping of fate-modifying co-factors in order to stratify differentiating cytotypes and predict lineage competency. Here, Flk-1-based selection of early embryonic stem cell progeny separated a population depleted of pluripotent (
Oct4,
Sox2) and endoderm (
Sox17) markers. The gene expression profile of the Flk-1
+ population was notable for a significant upregulation in the vasculogenic
Sox7 transcription factor, which overlapped with the emergence of primordial cardiac transcription factors
GATA-4,
Myocardin and
Nkx2.5. Sorting the parental Flk-1
+ pool with the chemokine receptor CXCR4 to enrich the cardiopoietic subpopulation uncovered divergent
Sox7 expression, with a 7-fold induction in non-cardiac compared to cardiac progenitors. Bioinformatic resolution sequestered a framework of gene expression relationship between Sox transcription factor family members and the Flk-1/CXCR4 axes with significant integration of
β-catenin signaling. Thus, differential
Sox7 gene expression presents a novel biomarker profile, and possible regulatory switch, to distinguish cardiovascular pedigrees within Flk-1
+ multi-lineage progenitors.
The benefit of pharmacogenetic testing before starting drug therapy has been well documented for several single gene–drug combinations. However, the clinical utility of a pre-emptive genotyping ...strategy using a pharmacogenetic panel has not been rigorously assessed.
We conducted an open-label, multicentre, controlled, cluster-randomised, crossover implementation study of a 12-gene pharmacogenetic panel in 18 hospitals, nine community health centres, and 28 community pharmacies in seven European countries (Austria, Greece, Italy, the Netherlands, Slovenia, Spain, and the UK). Patients aged 18 years or older receiving a first prescription for a drug clinically recommended in the guidelines of the Dutch Pharmacogenetics Working Group (ie, the index drug) as part of routine care were eligible for inclusion. Exclusion criteria included previous genetic testing for a gene relevant to the index drug, a planned duration of treatment of less than 7 consecutive days, and severe renal or liver insufficiency. All patients gave written informed consent before taking part in the study. Participants were genotyped for 50 germline variants in 12 genes, and those with an actionable variant (ie, a drug–gene interaction test result for which the Dutch Pharmacogenetics Working Group DPWG recommended a change to standard-of-care drug treatment) were treated according to DPWG recommendations. Patients in the control group received standard treatment. To prepare clinicians for pre-emptive pharmacogenetic testing, local teams were educated during a site-initiation visit and online educational material was made available. The primary outcome was the occurrence of clinically relevant adverse drug reactions within the 12-week follow-up period. Analyses were irrespective of patient adherence to the DPWG guidelines. The primary analysis was done using a gatekeeping analysis, in which outcomes in people with an actionable drug–gene interaction in the study group versus the control group were compared, and only if the difference was statistically significant was an analysis done that included all of the patients in the study. Outcomes were compared between the study and control groups, both for patients with an actionable drug–gene interaction test result (ie, a result for which the DPWG recommended a change to standard-of-care drug treatment) and for all patients who received at least one dose of index drug. The safety analysis included all participants who received at least one dose of a study drug. This study is registered with ClinicalTrials.gov, NCT03093818 and is closed to new participants.
Between March 7, 2017, and June 30, 2020, 41 696 patients were assessed for eligibility and 6944 (51·4 % female, 48·6% male; 97·7% self-reported European, Mediterranean, or Middle Eastern ethnicity) were enrolled and assigned to receive genotype-guided drug treatment (n=3342) or standard care (n=3602). 99 patients (52 1·6% of the study group and 47 1·3% of the control group) withdrew consent after group assignment. 652 participants (367 11·0% in the study group and 285 7·9% in the control group) were lost to follow-up. In patients with an actionable test result for the index drug (n=1558), a clinically relevant adverse drug reaction occurred in 152 (21·0%) of 725 patients in the study group and 231 (27·7%) of 833 patients in the control group (odds ratio OR 0·70 95% CI 0·54–0·91; p=0·0075), whereas for all patients, the incidence was 628 (21·5%) of 2923 patients in the study group and 934 (28·6%) of 3270 patients in the control group (OR 0·70 95% CI 0·61–0·79; p <0·0001).
Genotype-guided treatment using a 12-gene pharmacogenetic panel significantly reduced the incidence of clinically relevant adverse drug reactions and was feasible across diverse European health-care system organisations and settings. Large-scale implementation could help to make drug therapy increasingly safe.
European Union Horizon 2020.
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