Using Penalized Regression to Uncover Peer Effects in the Spatial Autoregressive Model

We propose a technique for estimating the spatial weights matrix (SWM) of the spatial autoregressive model (SAR) using the least absolute shrinkage and selection operator (Lasso), first proposed by Tibshirani (1996). The SWM is typically assumed a priori as a known matrix of correlations among spatially correlated data, as it cannot be estimated using standard techniques due to overfitting. However, we use the Lasso to discover the most prominent spatial coefficients in the SWM and estimate them using feasible generalized least squares, while setting the relatively unimportant effects to zero. Furthermore, the Lasso solutions are optimized to minimize mean squared error over a grid of possible spatial lag parameters. Finally, we use the LARS-Lasso algorithm to compute an illustrative example of the technique

Cold dispase digestion of murine lungs improves recovery and culture of airway epithelial cells

Airway epithelial cells (AECs) play a key role in maintaining lung homeostasis, epithelium regeneration and the initiation of pulmonary immune responses. To isolate and study murine AECs investigators have classically used short and hot (1h 37°C) digestion protocols. Here, we present a workflow for efficient AECs isolation and culture, utilizing long and cold (20h 4°C) dispase II digestion of murine lungs. This protocol yields a greater number of viable AECs compared to an established 1h 37°C dispase II digestion. Using a combination of flow cytometry and immunofluorescent microscopy, we demonstrate that compared to the established method, the cold digestion allows for recovery of a 3-fold higher number of CD45-CD31-EpCAM+ cells from murine lungs. Their viability is increased compared to established protocols, they can be isolated in larger numbers by magnetic-activated cell sorting (MACS), and they result in greater numbers of distal airway stem cell (DASC) KRT5+p63+ colonies in vitro. Our findings demonstrate that temperature and duration of murine lung enzymatic digestion have a considerable impact on AEC yield, viability, and ability to form colonies in vitro. We believe this workflow will be helpful for studying lung AECs and their role in the biology of lung.</p

Prevalence of extracranial venous abnormalities: results from a sample of 586 multiple sclerosis patients

The aim of this study was to assess the prevalence of chronic cerebrospinal venous insufficiency in an unselected cohort of multiple sclerosis (MS) patients. A total of 586 patients with clinically defined MS underwent catheter venography of the internal jugular veins, brachiocephalic veins and azygos vein. The following findings were regarded as pathologic: no outflow, slowed outflow, reversal of flow direction, prestenotic dilation accompanied by impaired outflow, outflow through collaterals, intraluminal structures obstructing the vein, hypoplasia, agenesia or significant narrowing of the vein. Venous abnormalities were found in 563 patients (96.1%). Lesions in one vein were found in 43.5%, in two veins in 49.5%, and in three veins in 3.1% of patients. Venous pathologies in the right internal jugular vein were found in 64.0% of patients, in the left internal jugular vein in 81.7%, in the left brachiocephalic vein in 1.0%, and in the azygos vein in 4.9%. Venous pathologies were found to be highly associated with MS, yet the clinical relevance of this phenomenon remains to be establishe

The use of modern telemedicine technologies in an innovative optimal cardiac rehabilitation program for patients after myocardial revascularization: Concept and design of RESTORE, a randomized clinical trial

Despite proven efficacy of cardiac rehabilitation (CR) in reducing the all-cause mortality in patients after myocardial revascularization, the penetration of CR, due to patient-related factors and referral rates remains limited. To improve the outcomes, home-based tele-rehabilitation (TR) has been proposed recently. In theory TR enhances the effects of standard CR procedures due to implementation of an intelligent monitoring system designed to ensure optimal training through on-demand transmission of vital signs, aimed at motivating the patients through daily schedule reminders, setting daily goals and creating a platform for mutual feedback. Several meta-analyses assessing various studies comparing these two methods (CR and TR) have proven that they are at least equally effective, with some of the research showing superiority of TR. Although there was a small sample size, lack of long-term follow-up, reporting effects of TR itself, no integration with tools designed for coaching, motivating and promoting a healthy lifestyle constitutes an important limitation. The latter carries a hopeful prognosis for improvement when utilizing a broad-spectrum approach, especially with use of dedicated technological solutions exploiting the fact of a large and yet rapidly increasing penetration of smartphones, mobile PCs and tablets in the population. The above-mentioned findings worked as the basis and rationale for commencing the RESTORE project aimed at developing and delivering state-of-the-art, comprehensive TR for patients after myocardial revascularization and evaluating its molecular aspect in view of how it influences the atherosclerosis progression attenuation. This paper presents the current state and rationale behind the project based on up-to-date TR efficacy data

Binding of RNA Aptamers to Membrane Lipid Rafts: Implications for Exosomal miRNAs Transfer from Cancer to Immune Cells

Intraluminal vesicles (ILVs) are released into the extracellular space as exosomes after the fusion of multivesicular bodies (MVBs) with the plasma membrane. miRNAs are delivered to the raft-like region of MVB by RNA-binding proteins (RBPs). RNA loading into exosomes can be either through direct interaction between RNA and the raft-like region of the MVB membrane, or through interaction between an RBP&ndash;RNA complex with this raft-like region. Selection of RNA aptamers that bind to lipid raft region of liposomal membranes was performed using the selection-amplification (SELEX) method. The pool of RNA aptamers was isolated, and the binding of this pool to lipid-raft regions was demonstrated. Sequencing of clones from rafted liposome-eluted RNAs showed sequences apparently of independent origin. Bioinformatics analysis revealed the most frequent raft-motifs present within these sequences. Four raft RNA motifs, one of them an EXO motif, have been identified. These motifs appear to be most frequent both in the case of raft RNA aptamers and in the case of exosomal pro-tumoral miRNAs transferred from cancer cells to macrophages, natural killer cells and dendritic cells, thus suggesting that the selection for incorporation of these miRNAs into ILVs is based on their affinity to the raft-like region of the MVB membrane

Anaerobinių pokyčių slenkstis lyginant su galutiniais maksimalaus 2000 m irklavimo ergometru testo rezultatais – Lenkijos nacionalinės irkluotojų komandos 1997–2005 metais atvejis

Ciklinėse ištvermės sporto šakose, prie kurių priskiriamas ir irklavimas, anaerobinio slenksčio (AT 4) indeksas gali būti laikomas patikimu pasirengimo irklavimo sezonui lygio indikatoriumi (Beneke, 2000; Foster, 1995; Messonnier, 1997). Sisteminė indekso kontrolė padeda pastebėti pirmuosius nukrypimo nuo užsibrėžtų pasirengimo tikslų požymius, tai leistų daryti treniruotės plano pakeitimus (Fitts ir kt., 1996; Kraemer ir kt., 2004; Sharkey ir kt., 2006). Darbo tikslas – aptarti anaerobinio slenksčio lygį, kuris buvo nustatomas 1997–2005 metais irkluotojams (n = 16), besirengiantiems pasaulio čempionatui ir olimpinėms žaidynėms. Autoriai gautus duomenis taip pat palygino su 2000 m irklavimo ergometru „Concept II“ simuliacinio testo rezultatais. Straipsnyje pateikti rezultatai rodo, kad tarp AT 4, pasiekto didėjančiomis fizinėmis pastangomis ir 2000 m specialiojo simuliacinio testo metu, yra stiprus tarpusavio ryšysIn cyclical endurance sports disciplines, and rowing is numbered among them, the index of anaerobic threshold (AT 4) may be considered to be a reliable indicator of the level of preparations for a rowing season (Beneke 2000; Foster 1995; Messonnier 1997). Systematic control of the index helps to notice first signs of a deviation from the defined goal of preparations thus enabling to make adjustments to a training plan (Fitts et al., 1996; Kraemer et al., 2004; Sharkey et al., 2006). The purpose of the work is to present the level of anaerobic threshold observed during rowers’ (n=16) preparations for World Championship and the Olympic Games in years 1997-2005. The authors have also compared the above thresholds with the results obtained during the simulation test of 2000 meters on a rowing ergometer “Concept II”. Results presented in the article show that there is a strong correlation between AT 4 power achieved during progressive physical effort and the results of the simulation specialist test of 2000 metersVytauto Didžiojo universitetasŠvietimo akademij

Digestion type does not affect cellular oxidative stress based on LDHa (lactate dehydrogenase) levels.

Following either hot or cold digestion CD45-CD31-EpCAM+ cells were MACS sorted and RNA isolated using Trizol and chloroform-based RNA isolation. SYBR green qPCR was performed, with Rpl37a as endogenous control. N = 3–4. (TIF)</p

Cold dispase II digest improves the viability and yield of highly pure MACS sorted CD45^-CD31^-EpCAM⁺ cells.

(a) Representative gating strategy for hot and cold digestion following debris exclusion (FSC/SSC) and singlets (FSC-H/FSC-A) for analysis of viability of CD45-CD31-EpCAM+ AECs. (b) Comparison of frequency of viable FACS-sorted CD45-CD31-EpCAM+ through live/dead near-IR fixable dye. Unpaired t-test (n = 21–33), median ± min/max. (c) Comparison of cell yield between the digest methods after CD45, CD31 MACS depletion and EpCAM+ MACS selection. Quantification using haemocytometer and trypan blue live/dead exclusion. Unpaired t-test (n = 8), median ± min/max. (d) Evaluation of cell suspension purity after MACS sorting CD45-CD31-EpCAM+ cells using flow cytometry. Each data point corresponds to a single murine lung. (e) Validating the CD45-CD31-EpCAM+ cells identity. CD45-CD31-EpCAM+ cells also express CD49f and CD24. Presented gating followed debris exclusion (FSC/SSC), singlets gating (FSC-H/FSC-A), live cells gating (LIVE/DEAD fixable near-IR/FSC-H), CD45 and CD31 exclusion (CD45/CD31) and EpCAM+ gating (EpCAM/FSC-H).</p

Murine AECs can be passaged (P0-2) following cold digestion and CD45^-CD31^-EpCAM⁺ MACS sorting.

(a) Following cold digestion, murine AECs (CD34-CD31-EpCAM+) were MACS sorted and 2x105 cells were seeded into a 24-well plate and cultured in a Stemcell PneumaCult Ex-Plus medium with 10μM Y-27632, 3μM CHIR99021, 1μM A 83–01 for 14 days. Cells were passaged twice. Each time cells were detached using TrypLE and split 1:10 into a larger well plate starting from 24-well plate (P0), followed by 12-well plate (P1) and lastly a 6-well plate (P2). Once cells reached confluency at the end of passage 2, cells were lifted up using TrypLE and 1.4x106 live cells (trypan blue exclusion) were counted using haemocytometer. Six 10x objective phase-contrast single fields of view (3x2) were stitched together. Scale bar 400μm. N>2. (b) All cells express KRT5 (green) after three passages of CD45-CD31-EpCAM+ MACS sorted AECs. Scale bar, 300μm. (TIF)</p