Background and Aims One of the challenges of international alcohol research and policy is the variability in and lack of

The nucleus contains the genetic information of the cell and all of the regulatory factors that process the genome effectively. The genome is encapsulated by a dense, filamentous meshwork called the nucleoskeleton, which is located at the inner nuclear membrane. The components of the nucleoskeleton are involved in cellular signaling (Wilson and Berk, 2010), but they are also necessary for maintaining nuclear structure, preventing rupture of the nucleus under force and possibly assisting in force transduction (Wang et al., 2009). Here, we present the integrated mechanical structures of the nucleoskeleton, including lamin filaments, multisubunit proteins, short actin filaments and the genome. We also discuss the integration of mechanical elements from the cytoskeleton into the nucleoskeleton. Based on the mechanical contributions of the individual elements, we demonstrate how mutations in nuclear structures might impact force-dependent etiologies of disease. The accompanying poster aims to provide a translational overview between the cell biology of the nucleus and the biophysics of its underlying polymeric structures.

The authors demonstrate the operation of a nanoscale field-effect pH sensor engineered from a functionalized silicon nanowire. With this nanofabricated pH sensor, the change in the hydrogen ion concentration or the pH value of a solution can be detected by the corresponding change in the nanowire differential conductance with a resolution of ±5nS∕pH. Fabrication of selective side gates on the nanowire sensor allows field-effect control of the surface charge on the nanowire by controlling the accumulation of charge carriers with the side-gate voltage. A simple physical model is used to analyze the observed data and to quantify the dependence of the conductance on pH. The development of a nanoscale sensor with physically engineered gates offers the possibility of highly parallel labeling and detection of chemical and biological molecules with selective control of individual array elements.

Dynamical response of nanomechanical cantilever structures immersed in a viscous fluid is important to in vitro single-molecule force spectroscopy, biomolecular recognition of disease-specific proteins, and the study of microscopic protein dynamics. Here we study the stochastic response of biofunctionalized nanomechanical cantilever beams in a viscous fluid. Using the fluctuation-dissipation theorem we derive an exact expression for the spectral density of displacement and a linear approximation for resonance frequency shift. We find that in a viscous solution the frequency shift of the nanoscale cantilever is determined by surface stress generated by biomolecular interaction with negligible contributions from mass loading due to the biomolecules.

Hutchinson-Gilford progeria syndrome (HGPS) is a premature aging syndrome caused by the expression and accumulation of a mutant form of lamin A, Δ50 lamin A. As a component of the cell's nucleoskeleton, lamin A plays an important role in the mechanical stabilization of the nuclear envelope and in other nuclear functions. It is largely unknown how the characteristic 50 amino acid deletion affects the conformation of the mostly intrinsically disordered tail domain of lamin A. Here we perform replica exchange molecular dynamics simulations of the tail domain and determine an ensemble of semi-stable structures. Based on these structures we show that the ZMPSTE 24 cleavage site on the precursor form of the lamin A tail domain orients itself in such a way as to facilitate cleavage during the maturation process. We confirm our simulated structures by comparing the thermodynamic properties of the ensemble structures to in vitro stability measurements. Using this combination of techniques, we compare the size, heterogeneity of size, thermodynamic stability of the Ig-fold, as well as the mechanisms of force-induced denaturation. Our data shows that the Δ50 lamin A tail domain is more compact and displays less heterogeneity than the mature lamin A tail domain. Altogether these results suggest that the altered structure and stability of the tail domain can explain changed protein-protein and protein-DNA interactions and may represent an etiology of the disease. Also, this study provides the first molecular structure(s) of the lamin A tail domain, which is confirmed by thermodynamic tests.

BackgroundThe global prevalence of PASC is estimated to be present in 0·43 and based on the WHO estimation of 470 million worldwide COVID-19 infections, corresponds to around 200 million people experiencing long COVID symptoms. Despite this, its clinical features are not well-defined.MethodsWe collected retrospective data from 140 patients with PASC in a post-COVID-19 clinic on demographics, risk factors, illness severity (graded as one-mild to five-severe), functional status, and 29 symptoms and principal component symptoms cluster analysis. The Institute of Medicine (IOM) 2015 criteria were used to determine the Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) phenotype.FindingsThe median age was 47 years, 59.0% were female; 49.3% White, 17.2% Hispanic, 14.9% Asian, and 6.7% Black. Only 12.7% required hospitalization. Seventy-two (53.5%) patients had no known comorbid conditions. Forty-five (33.9%) were significantly debilitated. The median duration of symptoms was 285.5 days, and the number of symptoms was 12. The most common symptoms were fatigue (86.5%), post-exertional malaise (82.8%), brain fog (81.2%), unrefreshing sleep (76.7%), and lethargy (74.6%). Forty-three percent fit the criteria for ME/CFS, majority were female, and obesity (BMI > 30 Kg/m2) (P = 0.00377895) and worse functional status (P = 0.0110474) were significantly associated with ME/CFS.InterpretationsMost PASC patients evaluated at our clinic had no comorbid condition and were not hospitalized for acute COVID-19. One-third of patients experienced a severe decline in their functional status. About 43% had the ME/CFS subtype.

Endothelial cells are stimulated by shear stress throughout the vasculature and respond with changes in gene expression and by morphological reorganization. Mechanical sensors of the cell are varied and include cell surface sensors that activate intracellular chemical signaling pathways. Here, possible mechanical sensors of the cell including reorganization of the cytoskeleton and the nucleus are discussed in relation to shear flow. A mutation in the nuclear structural protein lamin A, related to Hutchinson Gilford progeria syndrome, is reviewed specifically since the mutation results in altered nuclear structure and stiffer nuclei; animal models also suggest significantly altered vascular structure. Nuclear and cellular deformation of endothelial cells in response to shear stress provides partial understanding of possible mechanical regulation in the microcirculation. Increasing sophistication of fluid flow simulations inside the vessel is also an emerging area relevant to the microcirculation since visualization in situ is difficult. This integrated approach to study -including medicine, molecular and cell biology, biophysics and engineering -provides a unique understanding of multi-scale interactions in the microcirculation.

Lamin proteins contribute to nuclear structure and function, primarily at the inner nuclear membrane. The posttranslational processing pathway of lamin A includes farnesylation of the C-terminus, likely to increase membrane association, and subsequent proteolytic cleavage of the C-terminus. Hutchinson Gilford progeria syndrome is a premature aging disorder wherein a mutant version of lamin A, Δ50 lamin A, retains its farnesylation. We report here that membrane association of farnesylated Δ50 lamin A tail domains requires calcium. Experimental evidence and molecular dynamics simulations collectively suggest that the farnesyl group is sequestered within a hydrophobic region in the tail domain in the absence of calcium. Calcium binds to the tail domain with an affinity KD ≈ 250 μM where it alters the structure of the Ig-fold and increases the solvent accessibility of the C-terminus. In 2 mM CaCl2, the affinity of the farnesylated protein to a synthetic membrane is KD ≈ 2 μM, as measured with surface plasmon resonance, but showed a combination of aggregation and binding. Membrane binding in the absence of calcium could not be detected. We suggest that a conformational change induced in Δ50 lamin A with divalent cations plays a regulatory role in the posttranslational processing of lamin A, which may be important in disease pathogenesis.