Vector copy number (VCN), an average quantification of transgene copies unique to a CAR T cell product, is a characteristic that must be reported prior to patient administration as high VCN increases ...the risk of insertional mutagenesis. Historically, VCN assessment in CAR T cell products has been performed via quantitative polymerase chain reaction (qPCR). qPCR is reliable along a broad range of concentrations, but quantification requires use of a standard curve and precision is limited. Digital PCR (dPCR) methods were developed for absolute quantification of target sequences by counting nucleic acid molecules encapsulated in discrete, volumetrically defined partitions. Advantages of dPCR compared to qPCR include simplicity, reproducibility, sensitivity, and lack of dependency on a standard curve for definitive quantification. In the present study, we describe a dPCR assay developed for analysis of the novel bicistronic CD19x22 CAR T cell construct. We compare its performance on both the Bio-Rad QX200 droplet digital PCR (ddPCR) system and the QIAGEN QIAcuity nanoplate-based digital PCR (ndPCR) system to qPCR. The primer/probe assay was validated with qPCR, ndPCR and ddPCR using patient samples from preclinical CAR T cell manufacturing production runs, as well as Jurkat cell subclones which stably express this bicistronic CAR construct. ddPCR confirmed the specificity of this assay to detect only the bicistronic CAR product. Additionally, our assay gave accurate, precise, and reproducible CAR T cell VCN measurements across qPCR, ndPCR, and ddPCR modalities. We demonstrate that digital PCR strategies can be utilized for absolute quantification of CAR transgenes and VCN measurements, with improved test-retest reliability, and that specific assays can be developed for detection of unique constructs.
A series of symmetrically bis‐4‐methoxybenzyl (4MB) N‐substituted 1,4‐diketopyrrolo3,4‐cpyrrole (DPP) derivatives have been synthesized. The 4MB unit makes the DPP core soluble, and shows subtle ...modification of up to 0.2 eV in ground and excited states of the core when compared with related alkyl derivatives. Absorption and emission spectroscopy, as well as electrochemical and computational methods have been employed to prove the importance of the peripheral aryl units on the donor/ acceptor properties of the molecules. The 4MB products are highly fluorescent (quantum yields approaching 100 % in solution), with a unique distribution of frontier states shown by spectroelectrochemistry. The solid‐state fluorescence correlates with the X‐ray crystal structures of the compounds, a Stokes shift of approximately 80 nm is seen for two of the compounds. The frontier energy levels show that this subtle substitutional change could be of future use in molecular energy level tailoring in these, and related, materials for organic (opto)electronics.
A clever twist: The functionalisation of π‐conjugated DPP dyes with substituents that are orthogonal to the dye's conjugated backbone enhance the fluorescence by at least 10 % and is shown to subtlety modulate the levels of the ground and excited states by up to 0.2 eV in solution. In the solid state, the Stokes shift of 80 nm in two cases is correlated with the solid‐state structures.
Summary
In the intravenous iron therapy to treat iron deficiency anaemia in patients undergoing major abdominal surgery (PREVENTT) trial, the use of intravenous iron did not reduce the need for blood ...transfusion or reduce patient complications or length of hospital stay. As part of the trial protocol, serum was collected at randomisation and on the day of surgery. These samples were analysed in a central laboratory for markers of iron deficiency. We performed a secondary analysis to explore the potential interactions between pre‐operative markers of iron deficiency and intervention status on the trial outcome measures. Absolute iron deficiency was defined as ferritin <30 μg.l−1; functional iron deficiency as ferritin 30–100 μg.l−1 or transferrin saturation < 20%; and the remainder as non‐iron deficient. Interactions were estimated using generalised linear models that included different subgroup indicators of baseline iron status. Co‐primary endpoints were blood transfusion or death and number of blood transfusions, from randomisation to 30 days postoperatively. Secondary endpoints included peri‐operative change in haemoglobin, postoperative complications and length of hospital stay. Most patients had iron deficiency (369/452 82%) at randomisation; one‐third had absolute iron deficiency (144/452 32%) and half had functional iron deficiency (225/452 50%). The change in pre‐operative haemoglobin with intravenous iron compared with placebo was greatest in patients with absolute iron deficiency, mean difference 8.9 g.l−1, 95%CI 5.3–12.5; moderate in functional iron deficiency, mean difference 2.8 g.l−1, 95%CI −0.1 to 5.7; and with little change seen in those patients who were non‐iron deficient. Subgroup analyses did not suggest that intravenous iron compared with placebo reduced the likelihood of death or blood transfusion at 30 days differentially across subgroups according to baseline ferritin (p = 0.33 for interaction), transferrin saturation (p = 0.13) or in combination (p = 0.45), or for the number of blood transfusions (p = 0.06, 0.29, and 0.39, respectively). There was no beneficial effect of the use of intravenous iron compared with placebo, regardless of the metrics to diagnose iron deficiency, on postoperative complications or length of hospital stay.
Polyomaviruses infect a diverse range of mammalian and avian hosts, and are associated with a variety of symptoms. However, it is unknown whether the viruses are found in all mammalian families and ...the evolutionary history of the polyomaviruses is still unclear. Here, we report the discovery of a novel polyomavirus in the European badger (Meles meles), which to our knowledge represents the first polyomavirus to be characterized in the family Mustelidae, and within a European carnivoran. Although the virus was discovered serendipitously in the supernatant of a cell culture inoculated with badger material, we subsequently confirmed its presence in wild badgers. The European badger polyomavirus was tentatively named Meles meles polyomavirus 1 (MmelPyV1). The genome is 5187 bp long and encodes proteins typical of polyomaviruses. Phylogenetic analyses including all known polyomavirus genomes consistently group MmelPyV1 with California sea lion polyomavirus 1 across all regions of the genome. Further evolutionary analyses revealed phylogenetic discordance amongst polyomavirus genome regions, possibly arising from evolutionary rate heterogeneity, and a complex association between polyomavirus phylogeny and host taxonomic groups.
Introduction/ Background
Traditionally, MM harbouring t(11;14) is considered standard risk disease. In the era of novel therapies, however, pts with t(11;14) are reported to have inferior outcomes to ...other standard risk pts. Evidence of response to the BCL-2 inhibitor, Venetoclax (Ven), has further increased interest in understanding outcomes of t(11;14) pts. In this study, we aimed to describe the historical outcomes of t(11;14) MM and response to Ven in a real-world cohort of Australian pts.
Methods
This was a retrospective, multicentre study conducted by members of the Australasian Leukaemia and Lymphoma Group, Myeloma Working Party. Cases were identified by interrogation of cytogenetics/FISH database from 2010 to 2019 inclusive. Baseline patient and disease characteristics, treatment exposure and outcomes were extracted from hospital medical records. Descriptive statistics, and survival analyses were performed as appropriate.
Results
Seventy-four pts median age 65 years (yrs), range (43-85) were identified across seven centres. 43% pts had ISS Stage III MM with 88% harbouring additional cytogenetic abnormalities, incl. 13% with gain in 1q and 12% with del(17p). The majority (81%) of pts received proteasome inhibitor (PI)-based 1 st line therapy with 60% having an upfront autologous stem cell transplant (ASCT) and 54% having immunomodulatory drug (IMiD)-based maintenance therapy. Two patients received an allogeneic stem cell transplant after ASCT. The overall response rate (ORR) was 86% with 38% achieving very good partial response (VGPR) or better. The median progression free survival-1 (PFS-1) was 1.91 yrs (95% CI 1.73-2.56) PI-based, n=60, PFS 1.84yrs (95% CI 1.61-2.41) vs IMiD-based, n=5, PFS 4.58yrs (95% CI 1.16-5.51), HR 0.68 p=0.45 The median overall survival (OS) was 5.35 yrs (95% CI 4.12-6.56). Second and third line therapy was predominantly IMiD-based (see Table 1) with recent introduction of anti-CD38 monoclonal antibodies (mAbs). Median PFS-2 was 0.77 yrs (95% CI 0.39-0.98) while median PFS-3 was 0.65 yrs (95% CI 0.34-1.16). Eleven pts (median 3 prior lines of therapy) were given Ven Six pts in combination with PI, three with PI and mAbs and two with dexamethasone. ORR to Ven was 55% with 45% achieving VGPR or better. Median PFS with Ven was 0.54 yrs (85% CI 0.05-2.17). Median PFS for patients with 1-4 lines (n=6) was 1.22 years and median PFS for patients with 5 lines or more (n=5) was 0.54 years, HR 0.56 (95% CI 0.12-2.5).
Conclusion
For pts harbouring t(11;14), the duration of response to PI-based 1 st line therapy is suboptimal even with the use of ASCT consolidation in the majority of patients. Despite multiple prior lines of therapy, Venetoclax shows promising results and every effort should be made for early identification of this cytogenetic lesion. Further studies are required to examine the impact of novel triplet and quadruplet combination therapy and use of venetoclax early in the disease course of this sub-group of patients.
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Talaulikar: Takeda: Honoraria, Research Funding; Amgen: Honoraria, Research Funding; Jansenn: Honoraria, Research Funding; Roche: Honoraria, Research Funding; EUSA Pharma: Honoraria, Research Funding. Gibbs: Janssen, Celgene, Amgen, Takeda, BMS and Pfizer: Consultancy, Honoraria; AbbVie: Consultancy.
Studies of closed-loop control (CLC) in patients with type 1 diabetes (T1D) consistently demonstrate improvements in glycemic control as measured by increased time-in-range (TIR) 70-180 mg/dL. ...However, clinical predictors of TIR in users of CLC systems are needed.
We analyzed data from 100 children aged 6-13 years with T1D using the Tandem Control-IQ CLC system during a randomized trial or subsequent extension phase. Continuous glucose monitor data were collected at baseline and during 12-16 weeks of CLC use. Participants were stratified into quartiles of TIR on CLC to compare clinical characteristics.
TIR for those in the first, second, third, and fourth quartiles was 54%, 65%, 71%, and 78%, respectively. Lower baseline TIR was associated with lower TIR on CLC (
= 0.69,
< 0.001). However, lower baseline TIR was also associated with greater improvement in TIR on CLC (
= -0.81,
< 0.001). During CLC, participants in the highest versus lowest TIR-quartile administered more user-initiated boluses daily (8.5 ± 2.8 vs. 5.8 ± 2.6,
< 0.001) and received fewer automated boluses (3.5 ± 1.0 vs. 6.0 ± 1.6,
< 0.001). Participants in the lowest (vs. the highest) TIR-quartile received more insulin per body weight (1.13 ± 0.27 vs. 0.87 ± 0.20 U/kg/d,
= 0.008). However, in a multivariate model adjusting for baseline TIR, user-initiated boluses and insulin-per-body-weight were no longer significant.
Higher baseline TIR is the strongest predictor of TIR on CLC in children with T1D. However, lower baseline TIR is associated with the greatest improvement in TIR. As with open-loop systems, user engagement is important for optimal glycemic control.
Leishmania parasites cause a broad spectrum of clinical disease. Here we report the sequencing of the genomes of two species of Leishmania: Leishmania infantum and Leishmania braziliensis. The ...comparison of these sequences with the published genome of Leishmania major reveals marked conservation of synteny and identifies only approximately 200 genes with a differential distribution between the three species. L. braziliensis, contrary to Leishmania species examined so far, possesses components of a putative RNA-mediated interference pathway, telomere-associated transposable elements and spliced leader-associated SLACS retrotransposons. We show that pseudogene formation and gene loss are the principal forces shaping the different genomes. Genes that are differentially distributed between the species encode proteins implicated in host-pathogen interactions and parasite survival in the macrophage.
Celotno besedilo
Dostopno za:
DOBA, IJS, IZUM, KILJ, NUK, PILJ, PNG, SAZU, UILJ, UKNU, UL, UM, UPUK
Background:
According to claims-based studies, a minority of MDS patients receive disease-modifying therapy. Unfortunately, these studies are limited by design in describing the shared ...decision-making process between MDS patients and their physicians in choosing supportive care, such as blood transfusions. Therefore, we surveyed and interviewed transfusion-dependent (TD) MDS patients and MDS physicians about the choice and use of supportive care, blood transfusions, and chemotherapy.
Methods:
Nearly 300 TD-MDS patients and MDS physicians were surveyed or interviewed under an IRB exempt protocol (WIRB). Semi-structured interviews of 15 patients and 10 providers were conducted between July and December of 2018 to inform survey development. Patients and providers within the AAMDSIF database were then invited via email and social media to participate in anonymous surveys. Survey respondents were entered into a lottery to receive a $50 gift card. Each interviewed patient was offered a $25 gift card and each provider interviewee was offered a $50 gift card. Survey 1 queried MDS patients who self-reported they had been TD, defined as 2 or more blood transfusions within a 2-month timespan, at some time since 2014. Between September 2018 and February 2019, 157 TD-MDS patients responded. Primary variables of interest included patient age, sex, use of disease-modifying therapies, use of chelating agents, and frequency of blood transfusions. Survey 2 queried providers who regularly manage TD-MDS patients (n=109). Nominal and ordinal variables were tested for possible association by the Goodman-Kruskal tau statistic.
Results:
Demographics and baseline disease characteristics of TD-MDS patients are shown in Table 1. In brief, the patient cohort had a median age of 69 years (range, 36-83), male 51%, and a greater proportion of low and low-intermediate IPSS risk. Nearly half of patients were unaware of their IPSS risk group. The majority (57%) of patients were TD within the past 2 months of answering the survey. Two-thirds of patients received care from an MDS specialist. MDS providers practiced in academic medical centers (61%), community hematology/oncology specialty practices (17%), community urban hospitals (10%), community suburban hospitals (5%), and rural community hospitals (4%), with some providers practicing in multiple locations. On a monthly basis, the majority (85%) of providers cared for up to 10 patients with TD-MDS. The highest ranked patient concerns about blood transfusions were transfusion reaction and iron overload. Concern for the more common risk of bacterial infection ranked 5th and was least often selected as the most important concern to patients. Approximately half of patients reported use of an iron chelating agent. On qualitative interview, most patients did not recall a discussion with their physician regarding the risks of blood transfusions, despite providers reporting they regularly educate patients and ask for informed consent. Further, although 98% of patients visited their MDS care provider at least quarterly, 48% stated they had not discussed alternatives to blood transfusion with their provider. Many patients held the sentiment there was “no choice to be made” in receiving blood transfusions. While providers said blood transfusions as primary management of MDS was rare, they reported they would recommend transfusions as stand-alone therapy in settings of patient preference for supportive therapy only, failure of ESAs, patient desire for transfusion-only therapy, and older patient age. Most patients (65%) ranked their quality of life positively, with 6% ranking excellent. The majority (58%) of patients drove themselves to their transfusion clinic, and 54% of patients traveled greater than 10 miles to these visits. Older age significantly associated with higher transfusion frequency (G-Test = 31.318, χ2 df = 18, p-value = 0.02644), but not with use of disease-modifying therapy. The presence of comorbidities was not associated with either transfusion frequency or active therapy. No significant association was found between patient income and either transfusion frequency, BMT, or administration of HMAs.
Conclusions:
MDS patients and physicians hold differing perceptions about the choice to receive and risks of receiving blood transfusions. Appraisal and optimization of the informed consent process between TD-MDS patients and physicians are needed.
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No relevant conflicts of interest to declare.
Genomic and biosocial research data about individuals is rapidly proliferating, bringing the potential for novel opportunities for data integration and use. The scale, pace and novelty of these ...applications raise a number of urgent sociotechnical, ethical and legal questions, including optimal methods of data storage, management and access. Although the open science movement advocates unfettered access to research data, many of the UK's longitudinal cohort studies operate systems of managed data access, in which access is governed by legal and ethical agreements between stewards of research datasets and researchers wishing to make use of them. Amongst other things, these agreements aim to respect the reasonable expectations of the research participants who provided data and samples, as expressed in the consent process. Arguably, responsible data management and governance of data and sample use are foundational to the consent process in longitudinal studies and are an important source of trustworthiness in the eyes of those who contribute data to genomic and biosocial research.
This paper presents an ethnographic case study exploring the foundational principles of a governance infrastructure for Managing Ethico-social, Technical and Administrative issues in Data ACcess (METADAC), which are operationalised through a committee known as the METADAC Access Committee. METADAC governs access to phenotype, genotype and 'omic' data and samples from five UK longitudinal studies.
Using the example of METADAC, we argue that three key structural features are foundational for practising responsible data sharing: independence and transparency; interdisciplinarity; and participant-centric decision-making. We observe that the international research community is proactively working towards optimising the use of research data, integrating/linking these data with routine data generated by health and social care services and other administrative data services to improve the analysis, interpretation and utility of these data. The governance of these new complex data assemblages will require a range of expertise from across a number of domains and disciplines, including that of study participants. Human-mediated decision-making bodies will be central to ensuring achievable, reasoned and responsible decisions about the use of these data; the METADAC model described in this paper provides an example of how this could be realised.
The approaches and tools of health promotion can be useful for civil society groups, local and national governments and multilateral organizations that are working to operationalize the 2030 agenda ...for sustainable development. Health promotion and sustainable development share several core priorities, such as equity, intersectoral approaches and sustainability, that help maximize their impact across traditional sectoral boundaries. In the Region of the Americas, each of these priorities has strong resonance because of prominent and long-standing health inequities that are proving resistant to interventions driven solely by the health sector. We describe several cases from the World Health Organization's (WHO) Region of the Americas in which the approaches and tools of health promotion, with a focus on cities, healthy settings and multisectoral collaboration, have been used to put the agenda into practice. We highlight areas where such approaches and tools can be applied effectively and provide evidence of the transformative potential of health promotion in efforts to achieve the sustainable development goals.
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