Using Evaluation to Foster NYCETP Goals: Case Studies and Intercampsus Collaboration

This article describes the use of case studies as part of the formative evaluation conducted for the New York Collaborative for Excellence in Teacher Preparation (NYCETP). While case studies are often conducted for evaluations by outside experts, consultants, or evaluators themselves, we developed a strategy for case studies that used NYCETP faculty to case-study each other. This strategy involved cross-campus collaboration and cross-discipline (Arts & Science and Education) collaboration, and thus actively supported one of the NYCETP goals. The case study strategy also included the development of a faculty (peer) review form for evaluation of documentation of new and revised courses. Procedures for case studies and examples of case study benefits for faculty and evaluators are also described

Autonomous bioluminescence imaging of single mammalian cells with the bacterial bioluminescence system.

Bioluminescence-based imaging of living cells has become an important tool in biological and medical research. However, many bioluminescence imaging applications are limited by the requirement of an externally provided luciferin substrate and the low bioluminescence signal which restricts the sensitivity and spatiotemporal resolution. The bacterial bioluminescence system is fully genetically encodable and hence produces autonomous bioluminescence without an external luciferin, but its brightness in cell types other than bacteria has, so far, not been sufficient for imaging single cells. We coexpressed codon-optimized forms of the bacterial luxCDABE and frp genes from multiple plasmids in different mammalian cell lines. Our approach produces high luminescence levels that are comparable to firefly luciferase, thus enabling autonomous bioluminescence microscopy of mammalian cells

The Sensitivity of Auditory-Motor Representations to Subtle Changes in Auditory Feedback While Singing

Singing requires accurate control of the fundamental frequency (F0) of the voice. This study examined trained singers’ and untrained singers’ (nonsingers’) sensitivity to subtle manipulations in auditory feedback and the subsequent effect on the mapping between F0 feedback and vocal control. Participants produced the consonant-vowel /ta/ while receiving auditory feedback that was shifted up and down in frequency. Results showed that singers and nonsingers compensated to a similar degree when presented with frequency-altered feedback (FAF); however, singers’ F0 values were consistently closer to the intended pitch target. Moreover, singers initiated their compensatory responses when auditory feedback was shifted up or down 6 cents or more, compared to nonsingers who began compensating when feedback was shifted up 26 cents and down 22 cents. Additionally, examination of the first 50 ms of vocalization indicated that participants commenced subsequent vocal utterances, during FAF, near the F0 value on previous shift trials. Interestingly, nonsingers commenced F0 productions below the pitch target and increased their F0 until they matched the note. Thus, singers and nonsingers rely on an internal model to regulate voice F0, but singers’ models appear to be more sensitive in response to subtle discrepancies in auditory feedback

Time and duration of chondrule formation: Constraints from 26Al-26Mg ages of individual chondrules

Chondrules from unequilibrated ordinary and carbonaceous chondrites belong to the oldest and most primitive materials from the early solar system and record chemical and isotopic signatures relating to their formation and evolution. These signatures allow tracing protoplanetary disk processes that eventually led to the formation of planetary building blocks and rocky planets. 26Al-26Mg ages based on mineral-mesostasis isochrons of 31 porphyritic ferromagnesian chondrules, that belong mainly to type-II, constrain the time of chondrule melting prior to incorporation into the respective chondrite parent bodies. For this study chondrules from the unequilibrated L, L(LL) and LL ordinary chondrites (UOCs) NWA 5206, NWA 8276, MET 96503, MET 00452, MET 00526, NWA 7936 and QUE 97008 were selected, which are of petrologic types 3.00-3.15 and were thus least metamorphosed after formation. Magnesium and Al isotopes were measured in-situ by Secondary Ion Mass Spectrometry (SIMS) using a CAMECA 1280 ims. 26Mg excess from in-situ decay of 26Al correlating with 27Al/24Mg has been detected in the mesostasis of all but one chondrule. The initial Al isotopic compositions (26Al/27Al)0 and 26Mg/24Mg ratios (d26Mg*0) deduced from internal mineral isochron regressions range from (9.5 ± 2.8) × 10-6 to (3.1 ± 1.2) × 10-6 and -0.020 ± 0.028‰ to 0.011 ± 0.039‰, respectively. The corresponding chondrule ages (∆tCAI), calculated relative to calcium-aluminum-rich inclusions (CAIs) using the canonical 26Al/27Al = (5.23 ± 0.13) × 10-5, are between 1.76_(-0.27)^(+0.36) and 2.92_(-0.34)^(+0.51) Ma and date the melt formation and thus primary chondrule formation from dust-like precursors or reprocessing of older chondrules. The age range agrees with those acquired with different short-lived chronometers and with published 26Al-26Mg ages, the majority of which were obtained for chondrules from the Bishunpur and Semarkona meteorites, although no chondrule with (26Al/27Al)0 > 10-5 was found. Chondrules in single chondrite samples or between different chondrite groups show no distinct age distributions. The initial 26Al/27Al of the oldest chondrules in the L(LL)/LL and L chondrite samples are identical within their 1σ uncertainties and yield a mean age of 1.99_(-0.08)^(+0.08) Ma and 1.81_(-0.10)^(+0.11) Ma, respectively. The oldest chondrules from six of the seven studied samples record a mean age of 1.94_(-0.06)^(+0.07) Ma. Since heating events in the protoplanetary disk could have partially reset the Al-Mg systematics in pre-existing chondrules and this would have shifted recorded 26Al-26Mg ages toward younger dates, the oldest mean age of 1.81_(-0.10)^(+0.11) Ma recorded in L chondrite chondrules is interpreted to date the rapid and punctuated onset of chondrule formation. The density distribution of chondrule ages from this study, which comprises the largest single dataset of OC chondrule ages, combined with published ages for chondrules from ordinary and carbonaceous chondrites reveals major age peaks for OC chondrules at 2.0 and 2.3 Ma. Chondrules in ordinary and carbonaceous chondrites formed almost contemporaneously (with a possible distinction between CC groups) in two chemically distinct reservoirs, probably in density-enriched regions at the edges of Jupiter’s orbit. The young formation ages of chondrules suggest that they do not represent precursors but rather by-products of planetesimal accretion

Enhanced incorporation of subnanometer tags into cellular proteins for fluorescence nanoscopy via optimized genetic code expansion

With few-nanometer resolution recently achieved by a new generation of fluorescence nanoscopes (MINFLUX and MINSTED), the size of the tags used to label proteins will increasingly limit the ability to dissect nanoscopic biological structures. Bioorthogonal (click) chemical groups are powerful tools for the specific detection of biomolecules. Through the introduction of an engineered aminoacyl–tRNA synthetase/tRNA pair (tRNA: transfer ribonucleic acid), genetic code expansion allows for the site-specific introduction of amino acids with “clickable” side chains into proteins of interest. Welldefined label positions and the subnanometer scale of the protein modification provide unique advantages over other labeling approaches for imaging at molecular-scale resolution. We report that, by pairing a new N-terminally optimized pyrrolysyl–tRNA synthetase (chPylRS2020) with a previously engineered orthogonal tRNA, clickable amino acids are incorporated with improved efficiency into bacteria and into mammalian cells. The resulting enhanced genetic code expansion machinery was used to label β-actin in U2OS cell filopodia for MINFLUX imaging with minimal separation of fluorophores from the protein backbone. Selected data were found to be consistent with previously reported high-resolution information from cryoelectron tomography about the cross-sectional filament bundling architecture. Our study underscores the need for further improvements to the degree of labeling with minimal-offset methods in order to fully exploit molecularscale optical three-dimensional resolution

MINFLUX nanoscopy delivers 3D multicolor nanometer resolution in cells.

The ultimate goal of biological super-resolution fluorescence microscopy is to provide three-dimensional resolution at the size scale of a fluorescent marker. Here we show that by localizing individual switchable fluorophores with a probing donut-shaped excitation beam, MINFLUX nanoscopy can provide resolutions in the range of 1 to 3 nm for structures in fixed and living cells. This progress has been facilitated by approaching each fluorophore iteratively with the probing-donut minimum, making the resolution essentially uniform and isotropic over scalable fields of view. MINFLUX imaging of nuclear pore complexes of a mammalian cell shows that this true nanometer-scale resolution is obtained in three dimensions and in two color channels. Relying on fewer detected photons than standard camera-based localization, MINFLUX nanoscopy is poised to open a new chapter in the imaging of protein complexes and distributions in fixed and living cells

Neuromuscular Blockade with Rocuronium Bromide Increases the Tolerance of Acute Normovolemic Anemia in Anesthetized Pigs

Background: The patient's individual anemia tolerance is pivotal when blood transfusions become necessary, but are not feasible for some reason. To date, the effects of neuromuscular blockade (NMB) on anemia tolerance have not been investigated. Methods: 14 anesthetized and mechanically ventilated pigs were randomly assigned to the Roc group (3.78 mg/kg rocuronium bromide followed by continuous infusion of 1 mg/kg/min, n = 7) or to the Sal group (administration of the corresponding volume of normal saline, n = 7). Subsequently, acute normovolemic anemia was induced by simultaneous exchange of whole blood for a 6% hydroxyethyl starch solution (130/0.4) until a sudden decrease of total body O-2 consumption (VO2) indicated a critical limitation of O-2 transport capacity. The Hb concentration quantified at this time point (Hb(crit)) was the primary end-point of the protocol. Secondary endpoints were parameters of hemodynamics, O-2 transport and tissue oxygenation. Results: Hb(crit) was significantly lower in the Roc group (2.4 +/- 0.5 vs. 3.2 +/- 0.7 g/dl) reflecting increased anemia tolerance. NMB with rocuronium bromide reduced skeletal muscular VO2 and total body O-2 extraction rate. As the cardiac index increased simultaneously, total body VO2 only decreased marginally in the Roc group (change of VO2 relative to baseline -1.7 +/- 0.8 vs. 3.2 +/- 1.9% in the Sal group, p < 0.05). Conclusion: Deep NMB with rocuronium bromide increases the tolerance of acute normovolemic anemia. The underlying mechanism most likely involves a reduction of skeletal muscular VO2. During acellular treatment of an acute blood loss, NMB might play an adjuvant role in situations where profound stages of normovolemic anemia have to be tolerated (e.g. bridging an unexpected blood loss until blood products become available for transfusion). Copyright (C) 2011 S. Karger AG, Base

Integrated geophysical-petrological modeling of lithosphere-asthenosphere boundary in central Tibet using electromagnetic and seismic data

We undertake a petrologically driven approach to jointly model magnetotelluric (MT) and seismic surface wave dispersion (SW) data from central Tibet, constrained by topographic height. The approach derives realistic temperature and pressure distributions within the upper mantle and characterizes mineral assemblages of given bulk chemical compositions as well as water content. This allows us to define a bulk geophysical model of the upper mantle based on laboratory and xenolith data for the most relevant mantle mineral assemblages and to derive corresponding predicted geophysical observables. One-dimensional deep resistivity models were derived for two groups of MT stations. One group, located in the Lhasa Terrane, shows the existence of an electrically conductive upper mantle layer and shallower conductive upper mantle layer for the other group, located in the Qiangtang Terrane. The subsequent one-dimensional integrated petrological-geophysical modeling suggests a lithosphere-asthenosphere boundary (LAB) at a depth of 80¿120 km with a dry lithosphere for the Qiangtang Terrane. In contrast, for the Lhasa Terrane the LAB is located at about 180 km but the presence of a small amount of water in the lithospheric mantle (<0.02 wt%) is required to fit the longest period MT responses. Our results suggest two different lithospheric configurations beneath the southern and central Tibetan Plateau. The model for the Lhasa Terrane implies underthrusting of a moderately wet Indian plate. The model for the Qiangtang Terrane shows relatively thick and conductive crust and implies thin and dry Tibetan lithosphere.Peer Reviewe

Electromagnetic-field distribution measurements in the soft s-ray range: full characterization of a soft x-ray laser beam

Includes bibliographical references (pages 183901-4).We report direct measurement of the electromagnetic-field spatial distribution in a neonlike Ar capillary discharge-driven soft x-ray laser beam. The wave front was fully characterized in a single shot using a Shack-Hartmann diffractive optics sensor. The wave front was observed to be dependent on the discharge pressure and capillary length, as a result of beam refraction variations in the capillary plasma. The results predict ~70% of the laser beam energy can be focused into an area 4 times the size of the diffraction-limited spot, reaching intensities of ~4 × 1013 W/cm2

Mouse tissue harvest-induced hypoxia rapidly alters the in vivo metabolome, between-genotype metabolite level differences, and 13C-tracing enrichments

OBJECTIVE: Metabolomics as an approach to solve biological problems is exponentially increasing in use. Thus, this a pivotal time for the adoption of best practices. It is well known that disrupted tissue oxygen supply rapidly alters cellular energy charge. However, the speed and extent to which delayed mouse tissue freezing after dissection alters the broad metabolome is not well described. Furthermore, how tissue genotype may modulate such metabolomic drift and the degree to which traced METHODS: By combined liquid chromatography (LC)- and gas chromatography (GC)-mass spectrometry (MS), we measured how levels of 255 mouse liver metabolites changed following 30-second, 1-minute, 3-minute, and 10-minute freezing delays. We then performed test-of-concept delay-to-freeze experiments evaluating broad metabolomic drift in mouse heart and skeletal muscle, differential metabolomic change between wildtype (WT) and mitochondrial pyruvate carrier (MPC) knockout mouse livers, and shifts in RESULTS: Our data demonstrate that delayed mouse tissue freezing after dissection leads to rapid hypoxia-driven remodeling of the broad metabolome, induction of both false-negative and false-positive between-genotype differences, and restructuring of CONCLUSIONS: Our findings provide a previously absent, systematic illustration of the extensive, multi-domain metabolomic changes occurring within the early minutes of delayed tissue freezing. They also provide a novel, detailed resource of mouse liver ex vivo, hypoxic metabolomic remodeling