Laminin chain expression by chick chondrocytes and mouse cartilaginous tissues in vivo and in vitro

We have observed that laminins are expressed in the chondrocytes of chick embryo sternum, mouse limb bud, and adult mouse knee joint by the methods of in situ hybridization, immunohistochemistry, Western blotting, and immunoprecipitation. From in situ hybridization using similar sized RNA probes for different mouse laminin chains, mRNAs for the alpha 1, alpha 2, beta 1, beta 2, and gamma 1 chains were expressed in the chondrocytes of chick embryo sternum, mouse limb bud, and the articular cartilage cap and epiphyseal growth plate of adult mouse knee joint. Through the use of chain-specific antibodies, staining for laminins was observed in the cytoplasm of chondrocytes from chick embryo sternum, mouse limb bud, and adult mouse knee joint. Western blot analysis confirmed the presence of laminin chains in the cells and sternal tissues. Cultured chick embryonic sternal chondrocytes expressed laminin mRNAs in the proliferating stage (2-3 days of culture) but the level increased in the aggregated cells during the maturation stage (5-7 days of culture). Comparable data were also obtained after immunostaining the cells. Thus, laminins are expressed in significant amounts by chondrocytes and may have an important role in cartilage development.     

Chemical characterization of different typologies of mucilaginous aggregates in the Northern Adriatic Sea

The chemical composition of mucilage aggregates found during summer 2000, 2001 and 2002 in the North Adriatic Sea depends on the nature of the organic matter during aggregation, on the environmental conditions of the site of formation and on the transformations during ageing. The mucilages were composed of organic matter, together with a significant inorganic fraction. Elemental analysis revealed 12.5-32.2% of organic carbon, 0-7.3% of inorganic carbon and 1.0-3.7% of nitrogen. The C(org)/N ratios of most aggregates were between 7.5 and 12.6, values close to those found in the suspended matter; higher ratios were found in large-size (>5 m) aggregates which are probably older. The content of carbohydrates and proteins determined in the aggregates, respectively, 15.4+/-8.9% and 7.9+/-4.8%, w/w, showed a prevalence of carbohydrates over proteins. Neutral carbohydrate analysis of purified polysaccharides from mucilage samples showed very similar signatures with high relative abundance of galactose and glucose. Humic, fulvic and humin substances extracted from the mucilages constitute an important fraction of the organic matter in the aggregates. The humin (a fraction insoluble in acidic and basic media) was present in all mucilage samples, indicating the refractory nature of a part of the organic matter in the mucilage. The iron and calcium could play a role during the aggregation process to form a complex with polysaccharides and humic fractions. The C(org)/N ratio 10+/-2 found in the humic acids extracted from the Adriatic aggregates disclosed a marine origin. The low phosphorus content and the high C(org)/P ratio found in the aggregates might depend from high bacteria activity or from the aggregation of organic fractions depleted of phosphorus. The principal inorganic species contained aluminium and silicon, part of which was of biogenic origin and was more significant in the offshore mucilage aggregates than in the coastal ones. The Si(biog)/C(org) ratio showed that diatoms were always present in the aggregates, although it cannot be established whether these are the producers or these develop within the aggregates.     

Thermodynamic and kinetic aspects of the electron transfer reaction of bovine cytochrome c immobilized on 4-mercaptopyridine and 11-mercapto-1-undecanoic acid films

Bovine cytochrome c (cyt c) was adsorbed on a polycrystalline gold electrode coated with 4-mercaptopyridine and 11-mercapto-1-undecanoic acid self-assembled monolayers (SAMs) and the thermodynamics and kinetics of the heterogeneous protein-electrode electron transfer (ET) reaction were determined by cyclic voltammetry. The E°′ values for the immobilized protein were found to be lower than those for the corresponding diffusing species. The thermodynamic parameters for protein reduction ( and ) indicate that the stabilization of the ferric state due to protein–SAM interaction is enthalpic in origin. The kinetic data suggest that a tunneling mechanism is involved in the ET reaction: the distance between the redox center of the protein and the electrode surface can be efficiently evaluated using the Marcus equation.     

Review of highway runoff characteristics: Comparative analysis and universal implications

This review interprets highway runoff characterization studies performed on different continents. The results are synthesized to discuss the historical trends, first flush effects of pollutants, pollutant form as dissolved vs. particulate, and to identify surrogate water quality parameters. The information presented in this review showed that: (1) variability has been observed in all quality parameters from each continent and among continents; (2) with a few exceptions the variability seems to be within the expected range; (3) inconsistent monitoring data as well as inconsistent quality assurance and quality control measures were reported among studies, which may be partially responsible for variability of water quality results; (4) compared with historic data, the concentration of total Pb decreased exponentially, which can mostly be credited to leaded gasoline phase-out regulation; (5) first flush effects of pollutants based on concentration have been reported consistently (however, mass first flush effects for pollutants have been reported inconsistently compared with concentration first flush effect); (6) most metal pollutants and phosphorus are present in both the particulate and dissolved forms; and (7) strong correlations were observed between TSS, TDS, TOC and iron (Fe) and 13 other constituents and water quality parameters (turbidity, O&G, TPH, DOC, TKN, EC, Cl, Cd, Cr, Cu, Ni, Pb, Zn).     

An Efficient Aequorea victoria Green Fluorescent Protein for Stimulated Emission Depletion Super-Resolution Microscopy

In spite of their value as genetically encodable reporters for imaging in living systems, fluorescent proteins have been used sporadically for stimulated emission depletion (STED) super-resolution imaging, owing to their moderate photophysical resistance, which does not enable reaching resolutions as high as for synthetic dyes. By a rational approach combining steady-state and ultrafast spectroscopy with gated STED imaging in living and fixed cells, we here demonstrate that F99S/M153T/V163A GFP (c3GFP) represents an efficient genetic reporter for STED, on account of no excited state absorption at depletion wavelengths <600 nm and a long emission lifetime. This makes c3GFP a valuable alternative to more common, but less photostable, EGFP and YFP/Citrine mutants for STED imaging studies targeting the green-yellow region of the optical spectrum.     

A multi-level scheduler for batch jobs on grids

This paper proposes a two-level scheduler for dynamically scheduling a continuous stream of sequential and multi-threaded batch jobs on grids, made up of interconnected clusters of heterogeneous single-processor and/or symmetric multiprocessor machines. The scheduler aims to schedule arriving jobs respecting their computational and deadline requirements, and optimizing the hardware and software resource usage. At the top of the hierarchy a lightweight meta-scheduler (MS) classifies incoming jobs according to their requirements, and schedules them among the underlying resources balancing the workload. At cluster level a Flexible Backfilling algorithm carries out the job machine associations by exploiting dynamic information about the environment. Scheduling decisions at both levels are based on job priorities computed by using different sets of heuristics. The different proposals have been compared through simulations. Performance figures show the feasibility of our approach.     

Effects of positive end-expiratory pressure on right ventricular function in COPD patients during acute ventilatory failure

OBJECTIVE: To examine the effects of external positive end-expiratory pressure (PEEP) on right ventricular function in chronic obstructive pulmonary disease (COPD) patients with intrinsic PEEP (PEEPi). DESIGN: Prospective study. SETTING: General intensive care unit in a university teaching hospital. PATIENTS: Seven mechanically ventilated flow-limited COPD patients (PEEPi = 9.7 +/- 1.3 cmH2O, mean +/- SD) with acute respiratory failure. INTERVENTION: Hemodynamic and respiratory mechanic data were collected at four different levels of PEEP (0-5-10-15 cmH2O). MEASUREMENTS AND RESULTS: Hemodynamic parameters were obtained by a Swan-Ganz catheter with a fast response thermistor. Cardiac index (CI) and end-expiratory lung volume (EELV) reductions started simultaneously when the applied PEEP was approximately 90% of PEEPi measured on 0 cmH2O (ZEEP). Changes in transmural intrathoracic pressure (PEEPi,cw) started only at a PEEP value much higher (120%) than PEEPi. The reduction in CI was related to a decrease in the right end-diastolic ventricular volume index (RVEDVI) (r = 0.61; p < 0.001). No correlation between CI and transmural right atrial pressure was observed. The RVEDVI was inversely correlated with PEEP-induced changes in EELV (r = -55; p < 0.001), but no with PEEPi,cw (r = -0.08; NS). The relationship between RVEDVI and right ventricular stroke work index, considered an index of contractility, was significant in three patients, i.e., PEEP did not change contractility. In the other patients, an increase in contractility seemed to occur. CONCLUSIONS: In COPD patients an external PEEP exceeding 90% of PEEPi causes lung hyperinflation and reduces the CI due to a preload effect. The reduction in RVEDVI seems related to changes in EELV, rather than to changes in transmural pressures, suggesting a lung/heart volume interaction in the cardiac fossa. Thus, in COPD patients, application of an external PEEP level lower than PEEPi may affect right ventricular function.