Generating intravital super-resolution movies with conventional microscopy reveals actin dynamics that construct pioneer axons

Super-resolution microscopy is broadening our in-depth understanding of cellular structure. However, super-resolution approaches are limited, for numerous reasons, from utilization in longer-term intravital imaging. We devised a combinatorial imaging technique that combines deconvolution with stepwise optical saturation microscopy (DeSOS) to circumvent this issue and image cells in their native physiological environment. Other than a traditional confocal or two-photon microscope, this approach requires no additional hardware. Here, we provide an open-access application to obtain DeSOS images from conventional microscope images obtained at low excitation powers. We show that DeSOS can be used in time-lapse imaging to generate super-resolution movies in zebrafish. DeSOS was also validated in live mice. These movies uncover that actin structures dynamically remodel to produce a single pioneer axon in a 'top-down' scaffolding event. Further, we identify an F-actin population - stable base clusters - that orchestrate that scaffolding event. We then identify that activation of Rac1 in pioneer axons destabilizes stable base clusters and disrupts pioneer axon formation. The ease of acquisition and processing with this approach provides a universal technique for biologists to answer questions in living animals

Long-term outcome data from 121 patients treated with Gamma Knife stereotactic radiosurgery as salvage therapy for focally recurrent high-grade gliomas.

Introduction: We examined patient outcomes after Gamma Knife stereotactic radiosurgery (GKSRS) salvage therapy for recurrent high-grade gliomas (HGGs) to determine whether tumor grade or lesion size affected overall survival (OS) and progression-free survival (PFS). Methods: This single-center retrospective study assessed radiographic response and clinical outcomes following GKSRS salvage treatment of recurrent malignant gliomas (January 2005-March 2014). Results: A total of 121 patients (67 female) with 132 tumors were treated. Median (range) PFS was 4.7 (3.9-5.4) months for the cohort, 6.8 (4.6-8.9) months for initial grade 2 tumors, 4.2 (1.9-6.5) months for initial grade 3 tumors, and 4.3 (3.7-4.9) months for initial grade 4 tumors. Patients with small lesions (≤6.7 cm Conclusions: GKSRS offers meaningful salvage therapy with minimal morbidity in appropriately selected patients with focally recurrent HGGs

MicroRNA Function in Human Diseases

MicroRNAs are emerging as a hot topic in research, and rightfully so. They show great promise as targets of treatment and as markers for common human diseases, such as cancer and metabolic diseases. In this review, we address some of the basic questions regarding micro- RNA function in human disease and the clinical significance of microRNAs. Specifically, micro- RNAs in epigenetics, cancer, and metabolic diseases are discussed, with examples taken from cholangiocarcinoma and nonalcoholic fatty liver disease

MicroRNA Function in Human Diseases

MicroRNAs are emerging as a hot topic in research, and rightfully so. They show great promise as targets of treatment and as markers for common human diseases, such as cancer and metabolic diseases. In this review, we address some of the basic questions regarding micro- RNA function in human disease and the clinical significance of microRNAs. Specifically, micro- RNAs in epigenetics, cancer, and metabolic diseases are discussed, with examples taken from cholangiocarcinoma and nonalcoholic fatty liver disease

Observation of Quantum Effects in sub Kelvin Cold Reactions

There has been a long-standing quest to observe chemical reactions at low temperatures where reaction rates and pathways are governed by quantum mechanical effects. So far this field of Quantum Chemistry has been dominated by theory. The difficulty has been to realize in the laboratory low enough collisional velocities between neutral reactants, so that the quantum wave nature could be observed. We report here the first realization of merged neutral supersonic beams, and the observation of clear quantum effects in the resulting reactions. We observe orbiting resonances in the Penning ionization reaction of argon and molecular hydrogen with metastable helium leading to a sharp increase in the absolute reaction rate in the energy range corresponding to a few degrees kelvin down to 10 mK. Our method is widely applicable to many canonical chemical reactions, and will enable a breakthrough in the experimental study of Quantum Chemistry

Use of Complete-feed Diets RAMP™ and Test Starter for Receiving Cattle

Performance of newly arrived 576 lb steer calves, fed two complete feeds or a control ration was evaluated. Treatment diets were fed for 30 or 31 days and includeda control receiving diet consisting of alfalfa hay, Sweet Bran® , dry-rolled corn, and supplement or one of two complete feeds: RAMP and Test Starter which contained a high level of Sweet Bran and a minimal amount of forage. RAMP increased ADG when compared with the control diet. Cattle fed Test Starter had similar performance to the control receiving diet

High-speed, long-term, 4D in vivo lifetime imaging in intact and injured zebrafish and mouse brains by instant FLIM

Traditional fluorescence microscopy is blind to molecular microenvironment information that is present in the emission decay lifetime. With fluorescence lifetime imaging microscopy (FLIM), physiological parameters such as pH, refractive index, ion concentration, dissolved gas concentration, and fluorescence resonance energy transfer (FRET) can be measured. Despite these benefits, existing FLIM techniques are typically slow, noisy, and hard to implement due to expensive instrumentation and complex post-processing. To overcome these limitations, we present instant FLIM, a method that allows real-time acquisition and display of two-photon intensity, lifetime, and phasor imaging data. Using analog signal processing, we demonstrate in vivo four-dimensional (4D) FLIM movies by imaging mouse and zebrafish glial cell response to injury over 12 hours through intact skulls. Instant FLIM can be implemented as an upgrade to an existing multiphoton microscope using cost-effective off-the-shelf components, requires no data post-processing, and is demonstrated to be compatible with FD-FLIM super-resolution techniques

Concert recording 2021-11-15

[Track 1] Growth / Cole Powledge -- [Track 2] Day tripper ; Lady Madonna / Lennon/McCartney ; arranged by Tommy Emmanuel -- [Track 3] Disappear / Joshua Larson -- [Track 4] Sirabhorn / Pat Metheny -- [Track 5] Parisienne walkways / Gary Moore -- [Track 6] Songbird / Kenny G -- [Track 7] Drifting / Cody Lucas -- [Track 8] Slow dancing in a burning room / John Mayer -- [Track 9] Suites of morning

The Lamprey Genome: Illuminating Genomic Change across Eons and Embryogenesis

The lamprey genome provides unique insights into both the deep evolutionary history of vertebrate genomes and the maintenance of genome structure/integrity over development. The lamprey lineage diverged from all other vertebrates approximately 500 million years ago. As such, comparisons between lamprey and other vertebrates permit reconstruction of ancient duplication and rearrangement events that defined the fundamental architecture and gene content of all extant vertebrate genomes. Lamprey also undergoes programmatic changes genome structure that result in the physical elimination of ~20% of its genomic DNA (~0.5Gb from a ~2 Gb genome) from all somatic cell lineages during early embryonic development. Here, we outline recent progress in assembly and analysis of the lamprey germline genome, and progress in the development of methods for characterizing the cellular events that mediate DNA elimination. We have integrated information from several sampling approaches and sequencing technologies to achieve a highly contiguous assembly of lamprey genome (including: Illumina fragments/mate pairs, 20X coverage in Pacific Biosciences reads, dense meiotic maps and optical mapping data). This genome assembly has dramatically improved our ability to dissect the molecular basis and genetic outcomes of programmed genome rearrangements (PGRs), and has improved our understanding of the tempo and mode of large-­scale duplications and translocations within the ancestral vertebrate lineage. Analysis of the germline genome identifies several genes that are expressed in germline but physically eliminated from all somatic tissues. These eliminated genes correspond to several known oncogenes and appear to identify several other novel oncogene candidates. Complementing this assembly, the development of approaches to in situ analysis of 3D preserved cells has revealed that PGR unfolds through a series of dramatic cellular events that involve the programmatic alteration of several fundamental mechanisms of genome maintenance, including: alignment of chromosomes at metaphase, chromatid cohesion, separation and segregation, and nuclear envelope formation