Minimally invasive determination of mRNA concentration in single living bacteria

Fluorescence correlation spectroscopy (FCS) has permitted the characterization of high concentrations of noncoding RNAs in a single living bacterium. Here, we extend the use of FCS to low concentrations of coding RNAs in single living cells. We genetically fuse a red fluorescent protein (RFP) gene and two binding sites for an RNA-binding protein, whose translated product is the RFP protein alone. Using this construct, we determine in single cells both the absolute [mRNA] concentration and the associated [RFP] expressed from an inducible plasmid. We find that the FCS method allows us to reliably monitor in real-time [mRNA] down to ∼40 nM (i.e. approximately two transcripts per volume of detection). To validate these measurements, we show that [mRNA] is proportional to the associated expression of the RFP protein. This FCS-based technique establishes a framework for minimally invasive measurements of mRNA concentration in individual living bacteria

NUTMEG: A randomized phase II study of nivolumab and temozolomide versus temozolomide alone in newly diagnosed older patients with glioblastoma.

BACKGROUND: There is an immunologic rationale to evaluate immunotherapy in the older glioblastoma population, who have been underrepresented in prior trials. The NUTMEG study evaluated the combination of nivolumab and temozolomide in patients with glioblastoma aged 65 years and older. METHODS: NUTMEG was a multicenter 2:1 randomized phase II trial for patients with newly diagnosed glioblastoma aged 65 years and older. The experimental arm consisted of hypofractionated chemoradiation with temozolomide, then adjuvant nivolumab and temozolomide. The standard arm consisted of hypofractionated chemoradiation with temozolomide, then adjuvant temozolomide. The primary objective was to improve overall survival (OS) in the experimental arm. RESULTS: A total of 103 participants were randomized, with 69 in the experimental arm and 34 in the standard arm. The median (range) age was 73 (65-88) years. After 37 months of follow-up, the median OS was 11.6 months (95% CI, 9.7-13.4) in the experimental arm and 11.8 months (95% CI, 8.3-14.8) in the standard arm. For the experimental arm relative to the standard arm, the OS hazard ratio was 0.85 (95% CI, 0.54-1.33). In the experimental arm, there were three grade 3 immune-related adverse events which resolved, with no unexpected serious adverse events. CONCLUSIONS: Due to insufficient evidence of benefit with nivolumab, the decision was made not to transition to a phase III trial. No new safety signals were identified with nivolumab. This complements the existing series of immunotherapy trials. Research is needed to identify biomarkers and new strategies including combinations

NUTMEG: A randomized phase II study of nivolumab and temozolomide versus temozolomide alone in newly diagnosed older patients with glioblastoma

BackgroundThere is an immunologic rationale to evaluate immunotherapy in the older glioblastoma population, who have been underrepresented in prior trials. The NUTMEG study evaluated the combination of nivolumab and temozolomide in patients with glioblastoma aged 65 years and older.MethodsNUTMEG was a multicenter 2:1 randomized phase II trial for patients with newly diagnosed glioblastoma aged 65 years and older. The experimental arm consisted of hypofractionated chemoradiation with temozolomide, then adjuvant nivolumab and temozolomide. The standard arm consisted of hypofractionated chemoradiation with temozolomide, then adjuvant temozolomide. The primary objective was to improve overall survival (OS) in the experimental arm.ResultsA total of 103 participants were randomized, with 69 in the experimental arm and 34 in the standard arm. The median (range) age was 73 (65-88) years. After 37 months of follow-up, the median OS was 11.6 months (95% CI, 9.7-13.4) in the experimental arm and 11.8 months (95% CI, 8.3-14.8) in the standard arm. For the experimental arm relative to the standard arm, the OS hazard ratio was 0.85 (95% CI, 0.54-1.33). In the experimental arm, there were three grade 3 immune-related adverse events which resolved, with no unexpected serious adverse events.ConclusionsDue to insufficient evidence of benefit with nivolumab, the decision was made not to transition to a phase III trial. No new safety signals were identified with nivolumab. This complements the existing series of immunotherapy trials. Research is needed to identify biomarkers and new strategies including combinations

Measurement of B - anti-B mixing at the Z

From more than 175 000 hadronic Z decays observed with the ALEPH detector at LEP, we select 823 events with pairs of leptons in the final state. From these we measure χ, the probability thata b hadron which is observed to decay originated as a hadron. We find χ=0.132−0.026+0.027

Heavy flavour production in Z decays

From an analysis of inclusive leptons in data collected by the ALEPH detector at LEP, we measure the fractions of b and c events in hadronic Z decays. The b fraction times semileptonic branching ratio is measured to be . Assuming a b semileptonic branching ratio of 0.102 ± 0.010 gives , in good agreement with the standard model prediction of 0.217. The c fraction times semileptonic branching ratio is measured to be . Assuming a c semileptonic branching ratio of 0.090 ± 0.013 gives , in agreement with the standard model prediction of 0.171

A search for new quarks and leptons from Z0 decay at LEP

A search for Z0 decays into pairs of possible new heavy quarks (t and b′), new heavy charged leptons (L±), stable heavy neutral leptons (νL) and unstable heavy neutral leptons (L0) is performed on data collected by the ALEPH detector corresponding to 11 550 events of Z0→hadrons. The limits on the masses of the heavy quarks are Mt > 45.8 GeV and Mb′ > 46.0 GeV, allowing for both charged-current and flavor-changing neutral-current decays of the b′. If an L± decays into a stable νL, then for MνL MνL. Finally, while the mass of the stable νL is excluded up to 42.7 GeV, the mass of the unstable L0 is excluded up to 45.7 GeV with the mixing parameters |UℓL0|2 down to 10−13 at this mass. For 25.0 GeV < ML0 < 42.7 GeV, all values of |UℓL0|2 are excluded. All limits are given at 95% CL