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- Beal, Jacob, et al.
(författare)
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Robust estimation of bacterial cell count from optical density
- 2020
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Ingår i: Communications Biology. - : Springer Science and Business Media LLC. - 2399-3642. ; 3:1
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Tidskriftsartikel (refereegranskat)abstract
- Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data.
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- Deb, S., et al.
(författare)
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SUPERIOR SVG: no touch saphenous harvesting to improve patency following coronary bypass grafting (a multi-Centre randomized control trial, NCT01047449)
- 2019
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Ingår i: Journal of Cardiothoracic Surgery. - : Springer Science and Business Media LLC. - 1749-8090. ; 14
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Tidskriftsartikel (refereegranskat)abstract
- BackgroundSingle centre studies support No Touch (NT) saphenous vein graft (SVG) harvesting technique. The primary objective of the SUPERIOR SVG study was to determine whether NT versus conventional (CON) SVG harvesting was associated with improved SVG patency 1year after coronary artery bypass grafting surgery (CABG).MethodsAdults undergoing isolated CABG with at least 1 SVG were eligible. CT angiography was performed 1-year post CABG. Leg adverse events were assessed with a questionnaire. A systematic review was performed for published NT graft patency studies and results aggregated including the SUPERIOR study results.ResultsTwo hundred and-fifty patients were randomized across 12-centres (NT 127 versus CON 123 patients). The primary outcome (study SVG occlusion or cardiovascular (CV) death) was not significantly different in NT versus CON (NT: 7/127 (5.5%), CON 13/123 (10.6%), p=0.15). Similarly, the proportion of study SVGs with significant stenosis or total occlusion was not significantly different between groups (NT: 8/102 (7.8%), CON: 16/107 (15.0%), p=0.11). Vein harvest site infection was more common in the NT patients 1month postoperatively (23.3% vs 9.5%, p<0.01). Including this study's results, in a meta-analysis, NT was associated with a significant reduction in SVG occlusion, Odds Ratio 0.49, 95% Confidence Interval 0.29-0.82, p=0.007 in 3 randomized and 1 observational study at 1year postoperatively.ConclusionsThe NT technique was not associated with improved patency of SVGs at 1-year following CABG while early vein harvest infection was increased. The aggregated data is supportive of an important reduction of SVG occlusion at 1year with NT harvesting.Trial registrationNCT01047449.
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- Romano, K. A., et al.
(författare)
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Gut Microbiota-Generated Phenylacetylglutamine and Heart Failure
- 2023
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Ingår i: Circulation-Heart Failure. - : Ovid Technologies (Wolters Kluwer Health). - 1941-3289 .- 1941-3297. ; 16:1
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Tidskriftsartikel (refereegranskat)abstract
- Background: The gut microbiota-dependent metabolite phenylacetylgutamine (PAGln) is both associated with atherothrombotic heart disease in humans, and mechanistically linked to cardiovascular disease pathogenesis in animal models via modulation of adrenergic receptor signaling.Methods: Here we examined both clinical and mechanistic relationships between PAGln and heart failure (HF). First, we examined associations among plasma levels of PAGln and HF, left ventricular ejection fraction, and N-terminal pro-B-type natriuretic peptide in 2 independent clinical cohorts of subjects undergoing coronary angiography in tertiary referral centers (an initial discovery US Cohort, n=3256; and a validation European Cohort, n=829). Then, the impact of PAGln on cardiovascular phenotypes relevant to HF in cultured cardiomyoblasts, and in vivo were also examined.Results: Circulating PAGln levels were dose-dependently associated with HF presence and indices of severity (reduced ventricular ejection fraction, elevated N-terminal pro-B-type natriuretic peptide) independent of traditional risk factors and renal function in both cohorts. Beyond these clinical associations, mechanistic studies showed both PAGln and its murine counterpart, phenylacetylglycine, directly fostered HF-relevant phenotypes, including decreased cardiomyocyte sarcomere contraction, and B-type natriuretic peptide gene expression in both cultured cardiomyoblasts and murine atrial tissue.Conclusions: The present study reveals the gut microbial metabolite PAGln is clinically and mechanistically linked to HF presence and severity. Modulating the gut microbiome, in general, and PAGln production, in particular, may represent a potential therapeutic target for modulating HF.Registration: URL: ; Unique identifier: NCT00590200 and URL: ; Unique identifier: DRKS00020915.
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- Hillyard, P. B., et al.
(författare)
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Carrier-density-dependent lattice stability in InSb
- 2007
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Ingår i: Physical Review Letters. - 0031-9007 .- 1079-7114. ; 98:12, s. 125501-
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Tidskriftsartikel (refereegranskat)abstract
- The ultrafast decay of the x-ray diffraction intensity following laser excitation of an InSb crystal has been utilized to observe carrier dependent changes in the potential energy surface. For the first time, an abrupt carrier dependent onset for potential energy surface softening and the appearance of accelerated atomic disordering for a very high average carrier density have been observed. Inertial dynamics dominate the early stages of crystal disordering for a wide range of carrier densities between the onset of crystal softening and the appearance of accelerated atomic disordering.
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- Lee, Ho-Hsien, et al.
(författare)
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Expression, purification and crystallization of CTB-MPR, a candidate mucosal vaccine component against HIV-1
- 2014
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Ingår i: IUCrJ. - 2052-2525. ; 1:5, s. 305-317
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Tidskriftsartikel (refereegranskat)abstract
- CTB-MPR is a fusion protein between the B subunit of cholera toxin (CTB) andthe membrane-proximal region of gp41 (MPR), the transmembrane envelopeprotein ofHuman immunodeficiency virus 1(HIV-1), and has previously beenshown to induce the production of anti-HIV-1 antibodies with antiviralfunctions. To further improve the design of this candidate vaccine, X-raycrystallography experiments were performed to obtain structural informationabout this fusion protein. Several variants of CTB-MPR were designed,constructed and recombinantly expressed inEscherichia coli. The first variantcontained a flexible GPGP linker between CTB and MPR, and yielded crystalsthat diffracted to a resolution of 2.3 A ̊, but only the CTB region was detectedin the electron-density map. A second variant, in which the CTB was directlyattached to MPR, was shown to destabilize pentamer formation. A thirdconstruct containing a polyalanine linker between CTB and MPR proved tostabilize the pentameric form of the protein during purification. The purificationprocedure was shown to produce a homogeneously pure and monodispersesample for crystallization. Initial crystallization experiments led to pseudo-crystals which were ordered in only two dimensions and were disordered inthe third dimension. Nanocrystals obtained using the same precipitant showedpromising X-ray diffraction to 5 A ̊resolution in femtosecond nanocrystallo-graphy experiments at the Linac Coherent Light Source at the SLAC NationalAccelerator Laboratory. The results demonstrate the utility of femtosecondX-ray crystallography to enable structural analysis based on nano/microcrystalsof a protein for which no macroscopic crystals ordered in three dimensions havebeen observed before.
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- Lindenberg, A. M., et al.
(författare)
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X-ray diffuse scattering measurements of nucleation dynamics at femtosecond resolution.
- 2008
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Ingår i: Physical Review Letters. - 0031-9007 .- 1079-7114. ; 100:13, s. 135502-1-135502-5
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Tidskriftsartikel (refereegranskat)abstract
- Femtosecond time-resolved small and wide angle x-ray diffuse scattering techniques are applied to investigate the ultrafast nucleation processes that occur during the ablation process in semiconducting materials. Following intense optical excitation, a transient liquid state of high compressibility characterized by large-amplitude density fluctuations is observed and the buildup of these fluctuations is measured in real time. Small-angle scattering measurements reveal snapshots of the spontaneous nucleation of nanoscale voids within a metastable liquid and support theoretical predictions of the ablation process.
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- Stolte, W. C., et al.
(författare)
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Inner-shell photoionization of atomic chlorine
- 2013
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Ingår i: Physical Review A (Atomic, Molecular and Optical Physics). - 1050-2947. ; 88:5
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Tidskriftsartikel (refereegranskat)abstract
- Relative partial cross sections have been measured following photoexcitation of atomic chlorine near the Cl 2p and Cl 1s ionization thresholds. In addition, Breit-Pauli R-matrix calculations have been carried out in the region of the 2p thresholds, and the results are compared with experiment. Owing to angular-momentum considerations, it was found that the resonances associated with the higher 2p(-1) thresholds should be significantly wider than the lower ones, and this is borne out in both the experimental and the theoretical results. It is shown that a large number of resonance series contribute to the cross section, which make it difficult to untangle, and suggestions for further work to better understand the spectra are presented.
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- Buranawat, B, et al.
(författare)
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Evaluation of a Beta-Calcium Metaphosphate Bone Graft Containing Bone Morphogenetic Protein- 7 (OP-1) in Rabbit Maxillary Defects.
- 2014
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Ingår i: Journal of periodontology. - : Wiley. - 1943-3670 .- 0022-3492. ; 85:2, s. 298-307
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Tidskriftsartikel (refereegranskat)abstract
- Background: Calcium phosphate based materials have been widely used as bone substitutes and more recently are being exploited together with growth factors as bone tissue engineering scaffolds regulating cell behavior. The aim of the study was to evaluate the in vitro and in vivo response to a newly developed calcium metaphosphate bone graft, with and without bone stimulating growth factor. Materials and methods: Porous scaffolds of β calcium metaphosphate (CMP) were developed and extensively tested in vitro. Subsequently, CMP grafts with osteogenic protein 1 (OP-1) (test) and, without (control) were implanted into experimental rabbit maxillary bone defects. Animals were sacrificed 2, 4, and 8 weeks and samples examined with micro-computed tomography (µCT) and processed for histomorphometric analysis. Results: At 8 weeks the scaffolds containing OP-1 induced greater bone formation (P=0.018) than CMP alone, based on histomorphometric evaluation, % Bone area [Test 57.1±5.6 and Control 49.4±7.7] and micro-CT analysis, % Bone volume density (BV/TV) [Test 63.46±5.61 and Control 51.20±6.71]. Thus, this data indicated that both test and control CMP grafts showed a good degree of bone formation. Furthermore, the CMP materials showed signs of resorption from 4 weeks and no graft materials were observed at 8 weeks. Conclusion: In vitro the OP-1 loaded graft demonstrated a release profile and bioactivity over a 28 day period. In vivo testing confirmed enhanced bone formation of the OP-1 loaded graft after 8 weeks of healing.
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- Deb, Saswata, et al.
(författare)
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Composite vein grafting : Is it a "Y's" decision?
- 2015
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Ingår i: Journal of Thoracic and Cardiovascular Surgery. - New York, USA : Elsevier. - 0022-5223 .- 1097-685X. ; 149:2, s. 494-495
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Tidskriftsartikel (refereegranskat)
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16. |
- Deb, S., et al.
(författare)
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Individual variation in hemostatic alterations caused by tyrosine kinase inhibitors : a way to improve personalized cancer therapy?
- 2016
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Ingår i: Thrombosis Research. - : Elsevier. - 0049-3848 .- 1879-2472. ; 140:Suppl. 1, s. S196-S197
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Tidskriftsartikel (refereegranskat)abstract
- INTRODUCTION: During the last two decades, Bcr-Abl tyrosine kinase inhibitors (TKIs) have revolutionized the treatment of chronic myelogenous leukemia (CML), and are now considered standard treatment for this disease. However, TKIs can induce serious hemostatic side effects including cardiovascular disease and bleeding disorders. Blood platelet aggregation and formation of pro-coagulant platelets are important to allow a well-balanced hemostatic response. Therefore, a detailed understanding of what effect different TKIs exert on platelets and hemostasis could help to understand if there are differences of importance to minimize the risk of bleeding complications in treated patients.AIM: To investigate how TKIs used in CML (imatinib, dasatinib, nilotinib, bosutinib, and ponatinib) affect platelet activation and hemostasis.MATERIALS AND METHODS: We have developed a multi-parameter six color flow cytometry protocol to study different aspects of platelet function upon activation, e.g. formation of aggregatory (PAC-1-positive) and pro-coagulant (phosphatidylserine-exposing) platelets, exocytosis of alpha- and lysosomal granules and mitochondrial membrane potential.This protocol was performed in presence or absence of TKIs in blood from normal donors and in treated patients. Whole blood aggregometry (Multiplate®), thrombin generation in platelet-rich plasma and in vitro thrombus formation by free oscillation rheometry (ReoRox G2) was further evaluated in some situations.RESULTS: At clinically relevant concentrations, dasatinib significantly decreased the formation of procoagulant platelets. Ponatinib induced a slight decrease in formation of procoagulant platelets, whereas bosutinib and nilotinib showed opposite tendencies (n=7). Dasatinib also decreased platelet aggregation (n=4-6) and in vitro thrombus formation (n=3). Thrombin generation was not significantly affected by therapeutic levels of TKIs, whereas higher doses of dasatinib, bosutinib, ponatinib and imatinib significantly changed one or several of the thrombin generation parameters (n=7-8). Interestingly, large differences in response to the drugs were observed among the healthy donors, especially for dasatinib and bosutinib. Major inter-individual variations were also observed in dasatinib-treated patients, see Figure 1.CONCLUSIONS: Different TKIs show varying potency to affect platelet-based hemostasis. In addition, we found large inter-individual variations in how some drugs affected platelet function. Therefore, we suggest that development of a clinically useful protocol for platelet function testing could help to identify patients more susceptible to adverse drug reactions. Such a protocol could potentially help clinicians to gain insight into the risk of side effects, which could help to choose the most suitable drug for each individual patient.
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- Patra, Hirak Kumar, 1981-, et al.
(författare)
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Rational Nanotoolbox with Theranostic Potential for Medicated Pro-Regenerative Corneal Implants
- 2019
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Ingår i: Advanced Functional Materials. - : John Wiley & Sons. - 1616-301X .- 1616-3028. ; 29:38
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Tidskriftsartikel (refereegranskat)abstract
- Cornea diseases are a leading cause of blindness and the disease burden is exacerbated by the increasing shortage around the world for cadaveric donor corneas. Despite the advances in the field of regenerative medicine, successful transplantation of laboratory‐made artificial corneas is not fully realized in clinical practice. The causes of failure of such artificial corneal implants are multifactorial and include latent infections from viruses and other microbes, enzyme overexpression, implant degradation, extrusion or delayed epithelial regeneration. Therefore, there is an urgent unmet need for developing customized corneal implants to suit the host environment and counter the effects of inflammation or infection, which are able to track early signs of implant failure in situ. This work reports a nanotoolbox comprising tools for protection from infection, promotion of regeneration, and noninvasive monitoring of the in situ corneal environment. These nanosystems can be incorporated within pro‐regenerative biosynthetic implants, transforming them into theranostic devices, which are able to respond to biological changes following implantation.
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- Poli, Sven, et al.
(författare)
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Penumbral Rescue by normobaric O?=?O administration in patients with ischemic stroke and target mismatch proFile (PROOF): Study protocol of a phase IIb trial
- 2024
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Ingår i: INTERNATIONAL JOURNAL OF STROKE. - 1747-4930 .- 1747-4949. ; 19:1, s. 120-126
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Tidskriftsartikel (refereegranskat)abstract
- Rationale: Oxygen is essential for cellular energy metabolism. Neurons are particularly vulnerable to hypoxia. Increasing oxygen supply shortly after stroke onset could preserve the ischemic penumbra until revascularization occurs.Aims: PROOF investigates the use of normobaric oxygen (NBO) therapy within 6 h of symptom onset/notice for brain-protective bridging until endovascular revascularization of acute intracranial anterior-circulation occlusion.Methods and design: Randomized (1:1), standard treatment-controlled, open-label, blinded endpoint, multicenter adaptive phase IIb trial.Study outcomes: Primary outcome is ischemic core growth (mL) from baseline to 24 h (intention-to-treat analysis). Secondary efficacy outcomes include change in NIHSS from baseline to 24 h, mRS at 90 days, cognitive and emotional function, and quality of life. Safety outcomes include mortality, intracranial hemorrhage, and respiratory failure. Exploratory analyses of imaging and blood biomarkers will be conducted.Sample size: Using an adaptive design with interim analysis at 80 patients per arm, up to 456 participants (228 per arm) would be needed for 80% power (one-sided alpha 0.05) to detect a mean reduction of ischemic core growth by 6.68 mL, assuming 21.4 mL standard deviation.Discussion: By enrolling endovascular thrombectomy candidates in an early time window, the trial replicates insights from preclinical studies in which NBO showed beneficial effects, namely early initiation of near 100% inspired oxygen during short temporary ischemia. Primary outcome assessment at 24 h on follow-up imaging reduces variability due to withdrawal of care and early clinical confounders such as delayed extubation and aspiration pneumonia.
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- Rahman, Md Aminur, et al.
(författare)
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Arsenic
- 2023
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Ingår i: Inorganic Contaminants and Radionuclides. - : Elsevier. ; , s. 13-40
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Bokkapitel (övrigt vetenskapligt/konstnärligt)abstract
- Arsenic (As) is a poisonous metalloid and recognized as a Group I human carcinogen by the International Agency for Research on Cancer (IARC). Arsenic often exists in groundwater and surface water, oceanic and inland deposits, rocks, soils, and biota at variable concentrations. Over the last few decades, As contamination has been augmented noticeably due to both natural and anthropogenic sources. Arsenic contamination in groundwater is currently a major global environmental catastrophe, which affects over 200 million people in 107 countries and causes various health complications including cancer. Therefore, updated information regarding the sources, chemical form, bioavailability, extent and severity, food safety and regulation, remediation, and management of As is essential. In this chapter, we accumulated the detailed sources of As, including point and diffuse sources, various inorganic and organic As species, and their toxicity in the environment. Moreover, the fate of As in the environment, economic implications of As-contaminated food and food products, and the bioavailability and bio-accessibility of As in environmental media are also briefly summarized. Remediation technologies for As-contaminated soil with the latest case study and regulatory limits of As in soil are also presented in this chapter. Overall, this chapter incorporates the past and contemporary knowledge of As, which will be useful for better management of As in the near future.
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- Smith, Lucy K., et al.
(författare)
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An International Comparison of Death Classification at 22 to 25 Weeks' Gestational Age
- 2018
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Ingår i: Pediatrics. - : AMER ACAD PEDIATRICS. - 0031-4005 .- 1098-4275. ; 142:1
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVES: To explore international differences in the classification of births at extremely low gestation and the subsequent impact on the calculation of survival rates.METHODS: We used national data on births at 22 to 25 weeks' gestation from the United States (2014; n = 11144), Canada (2009-2014; n = 5668), the United Kingdom (2014-2015; n = 2992), Norway (2010-2014; n = 409), Finland (2010-2015; n = 348), Sweden (2011-2014; n = 489), and Japan (2014-2015; n = 2288) to compare neonatal survival rates using different denominators: all births, births alive at the onset of labor, live births, live births surviving to 1 hour, and live births surviving to 24 hours.RESULTS: For births at 22 weeks' gestation, neonatal survival rates for which we used live births as the denominator varied from 3.7% to 56.7% among the 7 countries. This variation decreased when the denominator was changed to include stillbirths (ie, all births [1.8%-22.3%] and fetuses alive at the onset of labor [3.7%-38.2%]) or exclude early deaths and limited to births surviving at least 12 hours (50.0%-77.8%). Similar trends were seen for infants born at 23 weeks' gestation. Variation diminished considerably at 24 and 25 weeks' gestation.CONCLUSIONS: International variation in neonatal survival rates at 22 to 23 weeks' gestation diminished considerably when including stillbirths in the denominator, revealing the variation arises in part from differences in the proportion of births reported as live births, which itself is closely connected to the provision of active care.
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- Tang, W. H. Wilson, et al.
(författare)
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Prognostic value of gut microbe-generated metabolite phenylacetylglutamine in patients with heart failure
- 2024
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Ingår i: EUROPEAN JOURNAL OF HEART FAILURE. - 1388-9842 .- 1879-0844. ; 26:2, s. 233-241
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Tidskriftsartikel (refereegranskat)abstract
- AimPhenylacetylglutamine (PAGln) is a phenylalanine-derived metabolite produced by gut microbiota with mechanistic links to heart failure (HF)-relevant phenotypes. We sought to investigate the prognostic value of PAGln in patients with stable HF.Methods and resultsFasting plasma PAGln levels were measured by stable-isotope-dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS) in patients with stable HF from two large cohorts. All-cause mortality was assessed at 5-year follow-up in the Cleveland cohort, and HF, hospitalization, or mortality were assessed at 3-year follow-up in the Berlin cohort. Within the Cleveland cohort, median PAGln levels were 4.2 (interquartile range [IQR] 2.4-6.9) mu M. Highest quartile of PAGln was associated with 3.09-fold increased mortality risk compared to lowest quartile. Following adjustments for traditional risk factors, as well as race, estimated glomerular filtration rate, amino-terminal pro-B-type natriuretic peptide, high-sensitivity C-reactive protein, left ventricular ejection fraction, ischaemic aetiology, and HF drug treatment, elevated PAGln levels remained predictive of 5-year mortality in quartile comparisons (adjusted hazard ratio [HR] [95% confidence interval, CI] for Q4 vs Q1: 1.64 [1.07-2.53]). In the Berlin cohort, a similar distribution of PAGln levels was observed (median 3.2 [IQR 2.0-4.8] mu M), and PAGln levels were associated with a 1.92-fold increase in 3-year HF hospitalization or all-cause mortality risk (adjusted HR [95% CI] for Q4 vs Q1: 1.92 [1.02-3.61]). Prognostic value of PAGln appears to be independent of trimethylamine N-oxide levels.ConclusionHigh levels of PAGln are associated with adverse outcomes independent of traditional cardiac risk factors and cardio-renal risk markers.
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- Verma, Subodh, et al.
(författare)
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Pedicled no-touch saphenous vein graft harvest limits vascular smooth muscle cell activation : the PATENT saphenous vein graft study
- 2014
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Ingår i: European Journal of Cardio-Thoracic Surgery. - : Oxford University Press. - 1010-7940 .- 1873-734X. ; 45:4, s. 717-725
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVE: Neointimal hyperplasia secondary to vascular smooth muscle cell (VSMC) activation limits the long-term patency of saphenous vein grafts (SVGs). We compared markers of vascular injury and VSMC activation in SVGs harvested using the pedicled 'no-touch' (NT) vs the conventional (CON) technique. METHODS: Patients undergoing coronary artery bypass surgery were enrolled in the PATENT SVG trial (clinicaltrials.gov NCT01488084). Patients were randomly allocated to have SVGs harvested with the NT technique from one leg and the CON method from the other. SVG segments underwent morphometry, histological and electron microscopy assessments and transcript measurements of VSMC activation and differentiation markers. Leg wound functional recovery and harvest site complications were assessed using a quality-of-life questionnaire. RESULTS: A total of 17 patients (65.3 +/- 7.3 years) were enrolled. SVGs harvested using the NT vs CON technique exhibited preserved intimal, medial and adventitial architecture. CON harvest was associated with greater medial Kruppel-like factor 4 transcript levels (0.26 +/- 0.05 vs 0.11 +/- 0.02, P < 0.05). CON samples had significantly lower medial serum response factor (0.53 +/- 0.11 vs 1.44 +/- 0.50, P < 0.05) and myocardin (0.59 +/- 0.08 vs 1.33 +/- 0.33, P < 0.05) transcript levels. MicroRNA-145, an inhibitor of VSMC activation and differentiation, was higher in the NT vs CON samples (1.84 +/- 1.03 vs 0.50 +/- 0.19, P < 0.05). Leg assessment scores were worse in the NT legs at 3 months, but similar to CON scores at 12 months. CONCLUSIONS: SVGs harvested using the 'NT' technique exhibit an early molecular and morphological pattern consistent with decreased VSMC activation compared with CON harvesting. Functional leg recovery was similar in both groups at 12 months. Larger studies are required to corroborate these findings.
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