Members of the cystatin superfamily interact with MMP-9 and protect it from autolytic degradation without affecting its gelatinolytic activities

Matrix metalloproteinases (MMPs), like other proteinases, can undergo autolytic degradation once activated in vivo. Whereas the activities of these enzymes are tightly regulated by tissue inhibitors of matrix metalloproteinases (TIMPs), it is not clear mechanistically how these enzymes are protected from autolysis in their active state. We previously reported that MMPs particularly MMP-9 and MMP-2 interact with the serum glycoprotein fetuin-A [Arch. Biochem. Biophys. (1995) 322, 250], a member of the cystatin superfamily. In the present analyses, we demonstrate that this interaction protects MMP-9 from autolytic degradation without interfering with its enzymatic activity, allowing it to efficiently digest gelatin. Our data demonstrate that MMP-9 binds to members of the cystatin family with K(diss) ranging from 25 to 58 nM for fetuin-A and 1.5-1.9 microM for cystatin C. The ability of fetuin-A to protect MMP-9 from autolysis requires a molar ratio of at least 8:1 (fetuin-A/MMP-9). More interestingly, our data show that the other members of the cystatin also have the ability to protect MMP-9 from autolysis, provided they are in molar excess relative to MMP-9. Taken together, our data suggest that cystatins, particularly fetuin-A, in any cellular compartment including the circulatory system, efficiently protect MMP-9 and possibly other MMPs from autolysis. This mechanism ensures the digestion of the preferred substrate for MMP-9 without sacrificing the enzyme in the process.     

Mechano-transduction mediated secretion and uptake of galectin-3 in breast carcinoma cells: implications in the extracellular functions of the lectin

In the following experiments, we sought to understand the triggering mechanism which propels galectin-3 to be secreted into the extracellular compartment from its intracellular stores in breast carcinoma cells. We also wanted to analyze in greater details the role of galectin-3 in cellular adhesion and spreading. To do this, we made use of two pairs of breast carcinoma cell lines where one of the pair has high expression of galectin-3 and the other low expression of the lectin. We determined that galectin-3 secreted into the conditioned medium of sub-confluent and spread cells in culture was quite low, almost negligible. However, once the cells were detached and rounded up, a mechano-sensing mechanism triggered the rapid secretion of galectin-3 into the conditioned medium. The secretion was constitutive as long as the cells remained detached. Galectin-3 was shown to be actively taken up from the conditioned medium by spreading cells. The cells which express and secrete high levels of galectin-3 adhered and spread much faster on plastic than those with reduced expression. The uptake of galectin-3 according to our data was important in cell spreading because if this process was compromised significantly, cells failed to spread. The data suggested that galectin-3 uptake modulates the adhesion plaques in that cells which express high levels of galectin-3 have thin-dot like plaques that may be suited for rapid adhesion and spreading while cells in which galectin-3 expression is reduced or knocked-down, have thick and elongated plaques which may be suited for a firmer adhesion to the substratum. Recombinant galectin-3 added exogenously reduced the thickness of the adhesion plaques of tumor cells with reduced galectin-3 expression. Taken together, the present data suggest that galectin-3 once externalized, is a powerful modulator of cellular adhesion and spreading in breast carcinoma cells.     

Status and distribution of mangrove forests of the world using earth observation satellite data

Aim Our scientific understanding of the extent and distribution of mangrove forests of the world is inadequate. The available global mangrove databases, compiled using disparate geospatial data sources and national statistics, need to be improved. Here, we mapped the status and distributions of global mangroves using recently available Global Land Survey (GLS) data and the Landsat archive. Methods We interpreted approximately 1000 Landsat scenes using hybrid supervised and unsupervised digital image classification techniques. Each image was normalized for variation in solar angle and earth–sun distance by converting the digital number values to the top‐of‐the‐atmosphere reflectance. Ground truth data and existing maps and databases were used to select training samples and also for iterative labelling. Results were validated using existing GIS data and the published literature to map ‘true mangroves’. Results The total area of mangroves in the year 2000 was 137,760 km² in 118 countries and territories in the tropical and subtropical regions of the world. Approximately 75% of world's mangroves are found in just 15 countries, and only 6.9% are protected under the existing protected areas network (IUCN I‐IV). Our study confirms earlier findings that the biogeographic distribution of mangroves is generally confined to the tropical and subtropical regions and the largest percentage of mangroves is found between 5° N and 5° S latitude. Main conclusions We report that the remaining area of mangrove forest in the world is less than previously thought. Our estimate is 12.3% smaller than the most recent estimate by the Food and Agriculture Organization (FAO) of the United Nations. We present the most comprehensive, globally consistent and highest resolution (30 m) global mangrove database ever created. We developed and used better mapping techniques and data sources and mapped mangroves with better spatial and thematic details than previous studies.     

Detachment of breast tumor cells induces rapid secretion of exosomes which subsequently mediate cellular adhesion and spreading

Exosomes are nano-vesicles secreted by a wide range of mammalian cell types. These vesicles are abundant in serum and other extracellular fluids and contain a large repertoire of proteins, mRNA and microRNA. Exosomes have been implicated in cell to cell communication, the transfer of infectious agents, and neurodegenerative diseases as well as tumor progression. However, the precise mechanisms by which they are internalized and/or secreted remain poorly understood. In order to follow their release and uptake in breast tumor cells in real time, cell-derived exosomes were tagged with green fluorescent protein (GFP)-CD63 while human serum exosomes were rhodamine isothiocynate-labeled. We show that detachment of adherent cells from various substrata induces a rapid and substantial secretion of exosomes, which then concentrate on the cell surfaces and mediate adhesion to various extracellular matrix proteins. We also demonstrate that disruption of lipid rafts with methyl-beta-cyclodextrin (MβCD) inhibits the internalization of exosomes and that annexins are essential for the exosomal uptake mechanisms. Taken together, these data suggest that cellular detachment is accompanied by significant release of exosomes while cellular adhesion and spreading are enhanced by rapid uptake and disposition of exosomes on the cell surface.     

Risk of Injection-Site Abscess among Infants Receiving a Preservative-Free,Two-Dose Vial Formulation of Pneumococcal Conjugate Vaccine in Kenya

There is a theoretical risk of adverse events following immunization with a preservative-free, 2-dose vial formulation of 10-valent-pneumococcal conjugate vaccine (PCV10). We set out to measure this risk. Four population-based surveillance sites in Kenya (total annual birth cohort of 11,500 infants) were used to conduct a 2-year post-introduction vaccine safety study of PCV10. Injection-site abscesses occurring within 7 days following vaccine administration were clinically diagnosed in all study sites (passive facility-based surveillance) and, also, detected by caregiver-reported symptoms of swelling plus discharge in two sites (active household-based surveillance). Abscess risk was expressed as the number of abscesses per 100,000 injections and was compared for the second vs first vial dose of PCV10 and for PCV10 vs pentavalent vaccine (comparator). A total of 58,288 PCV10 injections were recorded, including 24,054 and 19,702 identified as first and second vial doses, respectively (14,532 unknown vial dose). The risk ratio for abscess following injection with the second (41 per 100,000) vs first (33 per 100,000) vial dose of PCV10 was 1.22 (95% confidence interval [CI] 0.37–4.06). The comparator vaccine was changed from a 2-dose to 10-dose presentation midway through the study. The matched odds ratios for abscess following PCV10 were 1.00 (95% CI 0.12–8.56) and 0.27 (95% CI 0.14–0.54) when compared to the 2-dose and 10-dose pentavalent vaccine presentations, respectively. In Kenya immunization with PCV10 was not associated with an increased risk of injection site abscess, providing confidence that the vaccine may be safely used in Africa. The relatively higher risk of abscess following the 10-dose presentation of pentavalent vaccine merits further study.     

A simple framework for selection of water quality models

Water quality modeling is no longer just the preserve of specialists who seek to describe water quality processes but also for use by non specialists in everyday water quality management issues. With so many models already developed, it becomes prudent to adapt them to a situation than to develop a completely new model that would probably do the same simulations. The question is: which is the most appropriate model to apply to a situation? The specialist can always draw on past experiences to make a decision. However, this is not the case for the non specialist. A lot of different criteria can be used to decide which model to use for a particular situation based on some important factors. The objectives of modeling exercises differ and each water body is unique so there cannot be hard and fast rules on which is the best criteria for selecting the appropriate model. Furthermore, there is usually hardly any time on the project work plan allocated for model selection. Therefore there is need for a simple procedure to select the appropriate model. The objective of this paper was to develop a simple framework for selecting water quality models to aid the non specialist. The framework was then applied to a case study in order to evaluate its usefulness. The results from the case study show that after a thorough literature review, models can be evaluated against chosen criteria and the most appropriate model singled out. It was concluded that the framework is only effective if the research objective is adequately defined and the models are reviewed thoroughly, but it saves time for the actual modeling exercise.