Adhesion of flowing monocytes to hypoxia-reoxygenation-exposed endothelial cells: role of Rac1, ROS,and VCAM-1

Production ofreactive oxygen species (ROS) by ischemic tissue afterischemia-reperfusion (I/RP) is an important factor that contributes to tissue injury. The small GTPase Rac1 mediates the oxidative burst, and ROS act on signaling pathways involved in expression of inflammatory genes. Because there is evidence implicating monocytes in the pathogenesis of I/RP injury, our objective was todetermine the molecular mechanisms that regulate adhesive interactions between monocytes and hypoxia-reoxygenation (H/RO)-exposed cultured endothelial cells (ECs). When U937 cells were perfused over human umbilical vein ECs at 1 dyn/cm², H (1 h at 1%O₂)/RO (13 h) significantly increased the fluxes of rollingand stably adherent U937 cells. Either EC treatment with theantioxidant pyrrolidine dithiocarbamate (PDTC) or infection withAdRac1N17, which results in expression of the dominant-negative form ofRac1, abolished H/RO-induced ROS production, attenuated rolling, andabolished stable adhesion of U937 cells to H/RO-exposed ECs. Infectionwith AdRac1N17 also abolished H/RO-induced upregulation of vascularcell adhesion molecule (VCAM)-1. In turn, blocking VCAM-1 abolishedU937 cell stable adhesion and slightly increased rolling. We concludedthat the Rac1-dependent ROS partially regulate rolling and exclusivelyregulate stable adhesion of monocytic cells to ECs after H/RO and thatstable adhesion, but not rolling, is mediated by ROS-induced expressionof VCAM-1.

     

The initiation factor eIF4A is involved in the response to lithium stress in Saccharomyces cerevisiae

A gene, TIF2, was identified as corresponding to the translation initiation factor eIF4A and when overexpressed it confers lithium tolerance in galactose medium to Saccharomyces cerevisiae. Incubation of yeast with 6 mm LiCl in galactose medium leads to inhibition of [(35)S]methionine incorporation. By polysome analysis we show that translation is inhibited by lithium at the initiation step, accumulating 80 S monosomes. We further show by immunoblot analysis that when cells are incubated with lithium eIF4A does not sediment with ribosomal subunits. Overexpression of TIF2 overcomes inhibition of protein synthesis and restores its sedimentation with the initiation complex. In vivo, eIF4A is induced by lithium stress. We have shown previously that lithium is highly toxic to yeast when grown in galactose medium mainly due to inhibition of phosphoglucomutase, an enzyme responsible for the entry of galactose into glycolysis. We show that conditions that revert inhibition of phosphoglucomutase also revert inhibition of protein synthesis. Interestingly, glucose starvation leads to loss of polysomes but not to dissociation of eIF4A from the preinitiation complexes. Overexpression of SIT4, a protein phosphatase related to the TOR kinase pathway, reverts inhibition of protein synthesis by lithium and association of eIF4A with the initiation complex.     

Combined effects of air and soil pollution by fluoride emissions on Tibouchina pulchra Cogn., at Cubatão,SE Brazil,and their relations with aluminium

High deposition of gaseous/particulate fluorides and other air pollutants has resulted in an acidification and probable formation of soluble AlF_x complexes in the soil in the vicinity of the industrial complex of Cubatão, SE Brazil. With the present field study we aimed at determining the contribution of F and Al uptake from fluoride-contaminated soil, supposedly as AlF_x complexes, to the increase of foliar F and Al contents in saplings of an Al-accumulator tree species (Tibouchina pulchra) which were concomitantly exposed to fluoride-contaminated air and also the proportional contribution of both air and soil contamination to the mentioned foliar accumulation of these elements. The seasonal variations in F and Al accumulation and possible metabolic changes in the plants due to F and Al accumulation were also investigated. The saplings were exposed during three consecutive periods of 16 weeks to: (a) air and soil from a reference site (PV_noF); (b) air or soil from two polluted sites (CM-high air pollution, low F and MV-high air pollution, high F); and (c) both air and soil from these polluted sites. After exposure, the changes in the foliar concentrations of F and Al, the relations between both element contents and their relationships with oxidative stress indicators were determined. The data were grouped in three matrices: PV_noF–CM_lwF and PV_noF–MV_hgF, taking in account the possible air/soil exposure combinations in each, and soil/air from all sites. The slight F accumulation in plants of PV_noF–CM_lwF matrix was a result of higher uptake from soil than from air (54 and 46%, respectively). At PV_noF–MV_hgF matrix, the extremely high F accumulation in leaves of T. pulchra could be attributed to the combination of both air and soil contamination (83 and 17%, respectively). T. pulchra always showed higher foliar Al concentrations than 1000 g g^–1 dry mass, mainly after exposure to air and soil of both polluted sites (CM_lwF and MV_hgF). A highly significant linear regression was estimated between molar Al and F contents, taking in account the data obtained for saplings of T. pulchra cultivated in the different soils and exposed to ambient air of PV_noF, suggesting that both elements were taken as Al–F complexes from soil. The uptake of fluorides from air and/or soil of MV_hgF caused significant metabolic changes in T. pulchra, but visible injury supposedly induced by fluorides were observed only when the foliar F contents surpassed 700 g g^–1 dry mass. On the contrary, Al did not cause any metabolic stress to the plants.     

Seasonal cycle of planktonic communities at Inhaca Island, southern Mozambique

Monthly plankton sampling was carried out at three stationson the west coast of Inhaca Island, southern Mozambique, fromAugust 1994 to August 1995. Sampling included water mass physicalparameters, nutrients, chlorophyll and zooplankton with netsof 125 and 330 µm pore aperture. Nutrient concentrationhas shown maxima during the summer months, where rain providesthe maximum outflow of rivers discharging into Maputo Bay. Followingthe nutrient peak, chlorophyll a has shown maxima around themonth of April, with another minor peak in September, when temperaturebegins to increase. Zooplankton densities followed closely thephytoplankton peaks, especially small herbivorous taxa and larvalstages, such as gastropod and bivalve larvae.     

Disentangling host,pathogen, and environmental determinants of a recently emerged wildlife disease: lessons from the first 15 years of amphibian chytridiomycosis research

The amphibian fungal disease chytridiomycosis, which affects species across all continents, recently emerged as one of the greatest threats to biodiversity. Yet, many aspects of the basic biology and epidemiology of the pathogen, Batrachochytrium dendrobatidis (Bd), are still unknown, such as when and from where did Bd emerge and what is its true ecological niche? Here, we review the ecology and evolution of Bd in the Americas and highlight controversies that make this disease so enigmatic. We explore factors associated with variance in severity of epizootics focusing on the disease triangle of host susceptibility, pathogen virulence, and environment. Reevaluating the causes of the panzootic is timely given the wealth of data on Bd prevalence across hosts and communities and the recent discoveries suggesting co‐evolutionary potential of hosts and Bd. We generate a new species distribution model for Bd in the Americas based on over 30,000 records and suggest a novel future research agenda. Instead of focusing on pathogen “hot spots,” we need to identify pathogen “cold spots” so that we can better understand what limits the pathogen''s distribution. Finally, we introduce the concept of “the Ghost of Epizootics Past” to discuss expected patterns in postepizootic host communities.     

Inaccuracy of Venous Point-of-Care Glucose Measurements in Critically Ill Patients: A Cross-Sectional Study

Introduction

Current guidelines and consensus recommend arterial and venous samples as equally acceptable for blood glucose assessment in point-of-care devices, but there is limited evidence to support this recommendation. We evaluated the accuracy of two devices for bedside point-of-care blood glucose measurements using arterial, fingerstick and catheter venous blood samples in ICU patients, and assessed which factors could impair their accuracy.

Methods

145 patients from a 41-bed adult mixed-ICU, in a tertiary care hospital were prospectively enrolled. Fingerstick, central venous (catheter) and arterial blood (indwelling catheter) samples were simultaneously collected, once per patient. Arterial measurements obtained with Precision PCx, and arterial, fingerstick and venous measurements obtained with Accu-chek Advantage II were compared to arterial central lab measurements. Agreement between point-of-care and laboratory measurements were evaluated with Bland-Altman, and multiple linear regression models were used to investigate interference of associated factors.

Results

Mean difference between Accu-chek arterial samples versus central lab was 10.7 mg/dL (95% LA -21.3 to 42.7 mg/dL), and between Precision PCx versus central lab was 18.6 mg/dL (95% LA -12.6 to 49.5 mg/dL). Accu-chek fingerstick versus central lab arterial samples presented a similar bias (10.0 mg/dL) but a wider 95% LA (-31.8 to 51.8 mg/dL). Agreement between venous samples with arterial central lab was the poorest (mean bias 15.1 mg/dL; 95% LA -51.7 to 81.9). Hyperglycemia, low hematocrit, and acidosis were associated with larger differences between arterial and venous blood measurements with the two glucometers and central lab. Vasopressor administration was associated with increased error for fingerstick measurements.

Conclusions

Sampling from central venous catheters should not be used for glycemic control in ICU patients. In addition, reliability of the two evaluated glucometers was insufficient. Error with Accu-chek Advantage II increases mostly with central venous samples. Hyperglycemia, lower hematocrit, acidosis, and vasopressor administration increase measurement error.     

Cytokine profile of autologous conditioned serum for treatment of osteoarthritis, <Emphasis Type="Italic">in vitro</Emphasis>effects on cartilage metabolism and intra-articular levels after injection

Introduction  

Intraarticular administration of autologous conditioned serum (ACS) recently demonstrated some clinical effectiveness in treatment of osteoarthritis (OA). The current study aims to evaluate the in vitro effects of ACS on cartilage proteoglycan (PG) metabolism, its composition and the effects on synovial fluid (SF) cytokine levels following intraarticular ACS administration.     

Lipid-based colloidal carriers for peptide and protein delivery--liposomes versus lipid nanoparticles

This paper highlights the importance of lipid-based colloidal carriers and their pharmaceutical implications in the delivery of peptides and proteins for oral and parenteral administration. There are several examples of biomacromolecules used nowadays in the therapeutics, which are promising candidates to be delivered by means of liposomes and lipid nanoparticles, such as solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC). Several production procedures can be applied to achieve a high association efficiency between the bioactives and the carrier, depending on the physicochemical properties of both, as well as on the production procedure applied. Generally, this can lead to improved bioavailability, or in case of oral administration a more consistent temporal profile of absorption from the gastrointestinal tract. Advantages and drawbacks of such colloidal carriers are also pointed out. This article describes strategies used for formulation of peptides and proteins, methods used for assessment of association efficiency and practical considerations regarding the toxicological concerns.