Backscattering Mueller matrix polarimetry estimates microscale anisotropy and orientation in complex brain tissue structure.

Purpose: Diffusion magnetic resonance imaging (dMRI) quantitatively estimates brain microstructure, diffusion tractography being one clinically utilized framework. To advance such dMRI approaches, direct quantitative comparisons between microscale anisotropy and orientation are imperative. Complete backscattering Mueller matrix polarized light imaging (PLI) enables the imaging of thin and thick tissue specimens to acquire numerous optical metrics not possible through conventional transmission PLI methods.

Combined flat-field and frequency filter approach to correcting artifacts of multichannel two-photon microscopy.

Significance: Multiphoton microscopy (MPM) is a useful biomedical imaging tool for its ability to probe labeled and unlabeled depth-resolved tissue biomarkers at high resolution. Automated MPM tile scanning allows for whole-slide image acquisition but can suffer from tile-stitching artifacts that prevent accurate quantitative data analysis.

Aim: We have investigated postprocessing artifact correction methods using ImageJ macros and custom Python code.

Wide field-of-view fluorescence imaging for organ-level lineage tracing of rare intestinal stem cell populations.

Significance: Lineage tracing using fluorescent reporters is a common tool for monitoring the expression of genes and transcription factors in stem cell populations and their progeny. The zinc-binding protein 89 (ZBP-89/Zfp148 mouse gene) is a transcription factor that plays a role in gastrointestinal (GI) stem cell maintenance and cellular differentiation and has been linked to the progression of colon cancer. While lineage tracing is a useful tool, it is commonly performed with high-magnification microscopy on a small field of view within tissue sections, thereby limiting the ability to resolve reporter expression at the organ level.

Validation of a microwave energy meter to non-lethally estimate energetic reserves in adult sturgeon.

Whole-body (WB) energetic reserves influence fish survival, growth, and reproduction but are typically quantified using lethal methods ( proximate analyses) or interpreted through body condition indices. Energetic reserves can impact population dynamics through influences on growth rates, age-at-first-reproductive-maturity, and spawning periodicity at the individual-fish level, especially in long-lived sturgeon species. Therefore, a non-lethal tool to track the energetic reserves of endangered sturgeon populations could inform adaptive management and further our understanding of the sturgeon's biology.

Physical and cognitive impairments in people suffering from long COVID: protocol for a longitudinal population-based cohort study.

Introduction: Approximately 33% of people who contracted COVID-19 still experience symptoms 12 weeks after infection onset. This persistence of symptoms is now considered a syndrome itself called 'long COVID'. Evidence regarding long COVID and its cognitive and physical impacts is growing, but the literature is currently lacking objectively measured data to guide towards adapted healthcare trajectories.

Combined multiphoton microscopy and somatostatin receptor type 2 imaging of pancreatic neuroendocrine tumors.

Pancreatic neuroendocrine tumors (PNETs) are a rare but increasingly more prevalent cancer with heterogeneous clinical and pathological presentation. Surgery is the preferred treatment for all hormone-expressing PNETs and any PNET greater than 2 cm, but difficulties arise when tumors are multifocal, metastatic, or small in size due to lack of effective surgical localization. Existing techniques such as intraoperative ultrasound provide poor contrast and resolution, resulting in low sensitivity for such tumors.

Cognitive Performance Before and Following Habituation to Exercise-Induced Hypohydration of 2 and 4% Body Mass in Physically Active Individuals.

We investigated the effect of repeated exposures to hypohydration upon cognitive performance. In a randomized crossover design, ten physically active adults completed two 4-week training blocks, one where they maintained euhydration (EUH) and the other where they were water-restricted (DEH) during walking/running at 55% V.O2max, 40 °C.

Impact of Repeated Acute Exposures to Low and Moderate Exercise-Induced Hypohydration on Physiological and Subjective Responses and Endurance Performance.

This study aimed to examine whether repeated exposures to low (2%) and moderate (4%) exercise-induced hypohydration may reverse the potentially deleterious effect of hypohydration on endurance performance. Using a randomized crossover protocol, ten volunteers (23 years, V˙O: 54 mL∙kg∙min) completed two 4-week training blocks interspersed by a 5-week washout period. During one block, participants replaced all fluid losses (EUH) while in the other they were fluid restricted (DEH).

Gain of CTCF-Anchored Chromatin Loops Marks the Exit from Naive Pluripotency.

The genome of pluripotent stem cells adopts a unique three-dimensional architecture featuring weakly condensed heterochromatin and large nucleosome-free regions. Yet, it is unknown whether structural loops and contact domains display characteristics that distinguish embryonic stem cells (ESCs) from differentiated cell types. We used genome-wide chromosome conformation capture and super-resolution imaging to determine nuclear organization in mouse ESC and neural stem cell (NSC) derivatives.

Sex Differences in Mechanical Properties of the Achilles Tendon: Longitudinal Response to Repetitive Loading Exercise.

Lepley, AS, Joseph, MF, Daigle, NR, Digiacomo, JE, Galer, J, Rock, E, Rosier, SB, and Sureja, PB. Sex differences in mechanical properties of the Achilles tendon: Longitudinal response to repetitive loading exercise. J Strength Cond Res 32(11): 3070-3079, 2018-Sex differences have been observed in the mechanical properties of the Achilles tendon, which may help to explain the increased risk of injury in men.

Nuclear pore scaffold structure analyzed by super-resolution microscopy and particle averaging.

Much of life's essential molecular machinery consists of large protein assemblies that currently pose challenges for structure determination. A prominent example is the nuclear pore complex (NPC), for which the organization of its individual components remains unknown. By combining stochastic super-resolution microscopy, to directly resolve the ringlike structure of the NPC, with single particle averaging, to use information from thousands of pores, we determined the average positions of fluorescent molecular labels in the NPC with a precision well below 1 nanometer.

A system for imaging the regulatory noncoding Xist RNA in living mouse embryonic stem cells.

In mammals, silencing of one of the two X chromosomes in female cells compensates for the different number of X chromosomes between the sexes. The noncoding Xist RNA initiates X chromosome inactivation. Xist spreads from its transcription site over the X chromosome territory and triggers the formation of a repressive chromatin domain.

Intracellular transport by an anchored homogeneously contracting F-actin meshwork.

Actin-based contractility orchestrates changes in cell shape underlying cellular functions ranging from division to migration and wound healing. Actin also functions in intracellular transport, with the prevailing view that filamentous actin (F-actin) cables serve as tracks for motor-driven transport of cargo. We recently discovered an alternate mode of intracellular transport in starfish oocytes involving a contractile F-actin meshwork that mediates chromosome congression.

Normative data for the pyramids and palm trees test in the Quebec-French population.

Semantic memory tests assess long-term memory for facts, objects, and concepts as well as words and their meaning. Since it holds culturally shared information, the development of normative data adjusted to the cultural and linguistic reality of the target population is of particular importance. The present study aimed to establish normative data for the Pyramids and Palm Trees Test, a commonly used test of semantic memory, in the French-Quebec population.

Molecular crowding affects diffusion and binding of nuclear proteins in heterochromatin and reveals the fractal organization of chromatin.

The nucleus of eukaryotes is organized into functional compartments, the two most prominent being heterochromatin and nucleoli. These structures are highly enriched in DNA, proteins or RNA, and thus thought to be crowded. In vitro, molecular crowding induces volume exclusion, hinders diffusion and enhances association, but whether these effects are relevant in vivo remains unclear.

Nuclear import and assembly of influenza A virus RNA polymerase studied in live cells by fluorescence cross-correlation spectroscopy.

Intracellular transport and assembly of the subunits of the heterotrimeric RNA-dependent RNA polymerase constitute a key component of the replication cycle of influenza virus. Recent results suggest that efficient polymerase assembly is a limiting factor in the viability of reassortant viruses. The mechanism of nuclear import and assembly of the three polymerase subunits, PB1, PB2, and PA, is still controversial, yet it is clearly of great significance in understanding the emergence of new strains with pandemic potential.

Molecular characterization of a human cation-Cl- cotransporter (SLC12A8A, CCC9A) that promotes polyamine and amino acid transport.

Cation-Cl- cotransporters (CCCs) belong to a large family of proteins that includes 9 isoforms, two of which have still not been ascribed a transport function (CCC8 and CCC9) while the others are all known to promote Cl(-)-coupled Na+ and/or K+ movement at the cell surface. The CCCs are also included in a larger family termed amino acid-polyamine-organocation carriers (APCs). In contrast to the CCCs, however, polyamine (PA) transporters have thus far been isolated from unicellular species exclusively and do not all belong to the APC family.

LambdaN-GFP: an RNA reporter system for live-cell imaging.

We describe a GFP-based RNA reporter system (lambdaN-GFP) to visualize RNA molecules in live mammalian cells. It consists of GFP fused to an arginine-rich peptide derived from the phage lambda N protein, lambdaN22, which binds a unique minimal RNA motif and can be used to tag any RNA molecule. LambdaN-GFP uses a small and easy to engineer RNA tag, reducing the likelihood of perturbing the function of the tagged RNA molecule.

Structure and nuclear import function of the C-terminal domain of influenza virus polymerase PB2 subunit.

The trimeric influenza virus polymerase, comprising subunits PA, PB1 and PB2, is responsible for transcription and replication of the segmented viral RNA genome. Using a novel library-based screening technique called expression of soluble proteins by random incremental truncation (ESPRIT), we identified an independently folded C-terminal domain from PB2 and determined its solution structure by NMR. Using green fluorescent protein fusions, we show that both the domain and the full-length PB2 subunit are efficiently imported into the nucleus dependent on a previously overlooked bipartite nuclear localization sequence (NLS).

Dissecting the contribution of diffusion and interactions to the mobility of nuclear proteins.

Quantitative characterization of protein interactions under physiological conditions is vital for systems biology. Fluorescence photobleaching/activation experiments of GFP-tagged proteins are frequently used for this purpose, but robust analysis methods to extract physicochemical parameters from such data are lacking. Here, we implemented a reaction-diffusion model to determine the contributions of protein interaction and diffusion on fluorescence redistribution.

Novel insights regarding the operational characteristics and teleological purpose of the renal Na+-K+-Cl2 cotransporter (NKCC2s) splice variants.

The absorptive Na(+)-K(+)-Cl(-) cotransporter (NKCC2) is a polytopic protein that forms homooligomeric complexes in the apical membrane of the thick ascending loop of Henle (TAL). It occurs in at least four splice variants (called B, A, F, and AF) that are identical to one another except for a short region in the membrane-associated domain. Although each of these variants exhibits unique functional properties and distributions along the TAL, their teleological purpose and structural organization remain poorly defined.

Molecular mechanisms of cation transport by the renal Na+-K+-Cl- cotransporter: structural insight into the operating characteristics of the ion transport sites.

Two variants of the renal Na(+)-K(+)-Cl(-) cotransporter (NKCC2), called NKCC2A and NKCC2F, display marked differences in Na(+), Rb(+), and Cl(-) affinities, yet are identical to one another except for a 23-residue membrane-associated domain that is derived from alternatively spliced exons. The proximal portion of these exons is predicted to encode the second transmembrane domain (tm2) in the form of an alpha-helix, and the distal portion, part of the following connecting segment (cs1a). In recent studies, we have taken advantage of the A-F differences in kinetic behavior to determine which regions in tm2-cs1a are involved in ion transport.

A contractile nuclear actin network drives chromosome congression in oocytes.

Chromosome capture by microtubules is widely accepted as the universal mechanism of spindle assembly in dividing cells. However, the observed length of spindle microtubules and computer simulations of spindle assembly predict that chromosome capture is efficient in small cells, but may fail in cells with large nuclear volumes such as animal oocytes. Here we investigate chromosome congression during the first meiotic division in starfish oocytes.

LAP2alpha and BAF transiently localize to telomeres and specific regions on chromatin during nuclear assembly.

Lamina-associated polypeptide (LAP) 2alpha is a LEM (lamina-associated polypeptide emerin MAN1) family protein associated with nucleoplasmic A-type lamins and chromatin. Using live cell imaging and fluorescence microscopy we demonstrate that LAP2alpha was mostly cytoplasmic in metaphase and associated with telomeres in anaphase. Telomeric LAP2alpha clusters grew in size, formed 'core' structures on chromatin adjacent to the spindle in telophase, and translocated to the nucleoplasm in G1 phase.

Executive skills for medical faculty: a workshop description and evaluation.

As the healthcare system continues to change, healthcare professionals will need to assume an increasing number of administrative and management responsibilities. The goal of this article is to describe a two-day workshop on Executive Skills for Medical Faculty and the results of an evaluation conducted one year later. This workshop consisted of specific modules on analyzing time-management skills, determining goals and priorities, improving time-management strategies, assessing leadership styles and skills, and conducting effective meetings.