Upregulation of MMP-13 and TIMP-1 expression in response to mechanical strain in MC3T3-E1 osteoblastic cells

<p>Abstract</p> <p>Background</p> <p>Mechanical strain plays a significant role in the regulation of bone matrix turnover, which is mediated in part by matrix metalloproteinase (MMP)-13 and tissue inhibitors of matrix metalloproteinase (TIMP)-1. However, little is known about the correlation between mechanical strain and osteoblastic cell activities, including extracellular matrix (ECM) metabolism. Herein, we determined the effect of different magnitudes of cyclic tensile strain (0%, 6%, 12%, and 18%) on MMP-13 and TIMP-1 mRNA and protein expression in MC3T3-E1 osteoblasts. Furthermore, we employed specific inhibitors to examine the role of distinct signal transduction pathways known to mediate cellular responses to mechanical strain.</p> <p>Results</p> <p>We identified a magnitude-dependent increase in MMP-13 and TIMP-1 mRNA and protein levels in response to mechanical strains corresponding to 6%, 12%, and 18% elongation. The strain-induced increases in MMP-13 and TIMP-1 mRNA expression were inhibited by PD098059 and cycloheximide, respectively.</p> <p>Conclusions</p> <p>Our results suggest a mechanism for the regulation of bone matrix metabolism mediated by the differential expression of MMP-13 and TIMP-1 in response to increasing magnitudes of mechanical strain.</p

Climatic and volcanic forcing of tropical belt northern boundary over the past 800 years

The position of the northern boundary of the tropical belt affects the hydroclimate of many arid and semi-arid regions in the Northern Hemisphere. Widening of the tropical belt since the 1970s has largely been attributed to anthropogenic forcing. However, the relative influence of natural drivers of tropical belt expansion and contraction before this time is poorly understood. Here we use data on tree-ring widths from five mid-latitude regions in the Northern Hemisphere to reconstruct the movement of the northern boundary of the early spring tropical belt over the past 800 years (ad 1203–2003). Our reconstruction explains 45% of the interannual variance in the latitudinal extent of the Hadley circulation, a metric of the position of the tropical belt boundary. We find that the tropical belt contracted (expanded) during positive (negative) phases of the El Niño Southern Oscillation and Pacific North American teleconnection patterns. The tropical belt also contracted significantly following major volcanic events that injected sulfur into the stratosphere. The longest period of persistent tropical belt expansion occurred in the late sixteenth century, during one of the coldest periods of the Little Ice Age. Our results warn of potential socio-economic consequences of future variations in tropical belt width driven by natural climate variability or stratospheric aerosol injections, whether volcanic or artificial.Fil: Alfaro Sánchez, R.. Consejo Superior de Investigaciones Científicas. Centre de Recerca Ecológica I Aplicacions Forestals; España. University of Arizona; Estados UnidosFil: Nguyen, H.. Bureau of Meteorology; AustraliaFil: Klesse, S.. University of Arizona; Estados UnidosFil: Hudson, A.. University of Arizona; Estados UnidosFil: Belmecheri, S.. University of Arizona; Estados UnidosFil: Köse, N.. Istanbul University; TurquíaFil: Diaz, H. F.. University of Hawaii at Manoa; Estados UnidosFil: Monson, R. K.. University of Arizona; Estados UnidosFil: Villalba, Ricardo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Argentino de Nivología, Glaciología y Ciencias Ambientales. Provincia de Mendoza. Instituto Argentino de Nivología, Glaciología y Ciencias Ambientales. Universidad Nacional de Cuyo. Instituto Argentino de Nivología, Glaciología y Ciencias Ambientales; ArgentinaFil: Trouet, V.. University of Arizona; Estados Unido

Enabling adoption of 2D-NMR for the higher order structure assessment of monoclonal antibody therapeutics

The increased interest in using monoclonal antibodies (mAbs) as a platform for biopharmaceuticals has led to the need for new analytical techniques that can precisely assess physicochemical properties of these large and very complex drugs for the purpose of correctly identifying quality attributes (QA). One QA, higher order structure (HOS), is unique to biopharmaceuticals and essential for establishing consistency in biopharmaceutical manufacturing, detecting process-related variations from manufacturing changes and establishing comparability between biologic products. To address this measurement challenge, two-dimensional nuclear magnetic resonance spectroscopy (2D-NMR) methods were introduced that allow for the precise atomic-level comparison of the HOS between two proteins, including mAbs. Here, an inter-laboratory comparison involving 26 industrial, government and academic laboratories worldwide was performed as a benchmark using the NISTmAb, from the National Institute of Standards and Technology (NIST), to facilitate the translation of the 2D-NMR method into routine use for biopharmaceutical product development. Two-dimensional 1H,15N and 1H,13C NMR spectra were acquired with harmonized experimental protocols on the unlabeled Fab domain and a uniformly enriched-15N, 20%-13C-enriched system suitability sample derived from the NISTmAb. Chemometric analyses from over 400 spectral maps acquired on 39 different NMR spectrometers ranging from 500 MHz to 900 MHz demonstrate spectral fingerprints that are fit-for-purpose for the assessment of HOS. The 2D-NMR method is shown to provide the measurement reliability needed to move the technique from an emerging technology to a harmonized, routine measurement that can be generally applied with great confidence to high precision assessments of the HOS of mAb-based biotherapeutics