Development of an in vitro cell system from zebrafish suitable to study bone cell differentiation and extracellular matrix mineralization

Mechanisms of bone formation and skeletal development have been successfully investigated in zebrafish using a variety of in vivo approaches, but in vitro studies have been hindered due to a lack of homologous cell lines capable of producing an extracellular matrix (ECM) suitable for mineral deposition. Here we describe the development and characterization of a new cell line termed ZFB1, derived from zebrafish calcified tissues. ZFB1 cells have an epithelium-like phenotype, grow at 28 degrees C in a regular L-15 medium supplemented with 15% of fetal bovine serum, and are maintained and manipulated using standard methods (e.g., trypsinization, cryopreservation, and transfection). They can therefore be propagated and maintained easily in most cell culture facilities. ZFB1 cells show aneuploidy with 2n=78 chromosomes, indicative of cell transformation. Furthermore, because DNA can be efficiently delivered into their intracellular space by nucleofection, ZFB1 cells are suitable for gene targeting approaches and for assessing gene promoter activity. ZFB1 cells can also differentiate toward osteoblast or chondroblast lineages, as demonstrated by expression of osteoblast- and chondrocyte-specific markers, they exhibit an alkaline phosphatase activity, a marker of bone formation in vivo, and they can mineralize their ECM. Therefore, they represent a valuable zebrafish-derived in vitro system for investigating bone cell differentiation and extracellular matrix mineralization.FISHCELL project [PTDC/MAR/105313/2008]; Portuguese Science and Technology Foundation (FCT); European Regional Development Fund (ERDF) through the COMPETE Program; National Fund through FCT [PEst-C/MAR/LA0015/2011]; FCT [SFRH/BPD/39189/2007]; Association of European Marine Biological Laboratories through the ASSEMBLE project [FP7/227799]info:eu-repo/semantics/publishedVersio

Nations within a nation: variations in epidemiological transition across the states of India, 1990–2016 in the Global Burden of Disease Study

18% of the world's population lives in India, and many states of India have populations similar to those of large countries. Action to effectively improve population health in India requires availability of reliable and comprehensive state-level estimates of disease burden and risk factors over time. Such comprehensive estimates have not been available so far for all major diseases and risk factors. Thus, we aimed to estimate the disease burden and risk factors in every state of India as part of the Global Burden of Disease (GBD) Study 2016

Effect of atmospheric relative humidity on aerosol size distribution

175-188A method of accounting for the effect of variation in relative humidity on aerosol size distribution obtained from a low-pressure impactor (KPI) experiment is proposed. The size distribution of atmospheric aerosols under the prevailing relative humidities estimated by this method is compared with that obtained using a bistatic CW lidar. These results are used to study the effect of relative humidity on aerosol size distributions. At higher altitudes in the mixing region, increase in relative humidity causes a decrease in aerosol size index. The seasonal variation of aerosol number density near the surface is found to agree fairly well with the seasonal variation of the mixing region aerosol optical depth

A comparison of aerosol size distributions obtained from bistatic lidar and low-pressure impactor experiments at a coastal station

42-49Size distribution of aerosols in the atmospheric mixed region is studied using a bistatic CW lidar and a low-pressure impactor. Results obtained from these two experiments are compared. CW lidar observations showed that the size index (assuming the size distribution to follow a power law) generally lies in the range 3.5-5, whereas the size index obtained using the low-pressure impactor generally lies in the range 3.7-4.2

Isolation, Culture, and Differentiation of Blastema Cells from the Regenerating Caudal Fin of Zebrafish

The caudal fin of teleost fish has become an excellent system for investigating the mechanisms of epimorphic regeneration. Upon amputation of the caudal fin, a mass of undifferentiated cells, called blastema, proliferate beneath the wound-epidermis and differentiate into various cell types to faithfully restore the missing fin structures. Here we describe a protocol that can be used to isolate and culture blastema cells from zebrafish. Primary cultures were initiated from 36 h post-amputation (hpa) blastema and optimal cell growth was achieved using L-15 medium supplemented with 5% fetal bovine serum in plates either coated with fibronectin or uncoated. After seeding, zebrafish blastema cells formed a uniform culture and exhibited polygonal shapes with prominent nucleus, while various cell types were also observed after few days in culture indicating cell differentiation. Upon treatment with all-trans retinoic acid, zebrafish blastema cells differentiated into neuron-like and oligodendritic-like cells. Immunocytochemistry data also revealed the presence of mesenchymal and neuronal cells. The availability of blastema cell cultures could contribute to a better understanding of epimorphic regeneration by providing a mean to investigate the mechanisms underlying blastema cell differentiation. Furthermore, this protocol is simple, rapid, and cost-efficient, and can be virtually applied to the development of any fish blastema cell culture.</jats:p

Lidar observations on aerosol mixing height in a tropical coastal environment

15-21The aerosol mixing height is derived from the altitude distribution of aerosols in the lower atmosphere up to ~1.2 km during the post-sunset period (~2000 hrs IST) using a bi-static CW lidar at a tropical coastal station, Trivandrum. The mixing height shows significant seasonal variation with maxima in the dry months of March and November as well as in the south-west monsoon period. The maxima in March and November are attributed to strong convective mixing, whereas the maximum in the south-west monsoon is attributed to mechanical mixing due to strong wind shears