Effects of different forms of chitosan on intercellular junctions of mouse fibroblasts in vitro

Chitosan is a linear polysaccharide that has many biomedical applications. We compared the effects of chitosan, in both solution and membranous form, on intercellular adhesion of Swiss 3T3 mouse fibroblasts. Cells were grown as spheroidal cell cultures. Some control cell spheroids were cultured without chitosan and two experimental groups were cultured with chitosan. Chitosan in solution was used for one experimental group and chitosan in membranous form was used for the other. For each group, intercellular adhesion was investigated on days 5 and 10 of culture. Transmission electron microscopy revealed well-defined cellular projections that were more prominent in cells exposed to either membranous or solution forms of chitosan than to the chitosan-free control. Immunocytochemical staining of ICAM-1 and e-cadherin was used to determine the development of intercellular junctions. Compared to the weakly stained control, strong reactions were observed in both chitosan exposed groups at both 5 and 10 days. Cells were treated with 5-bromo-2-deoxyuridine (BrdU) and incubated with anti-BrdU primary antibody to assess proliferation. Both the solution and membranous forms of chitosan increased proliferation at both 5 and 10 days. Cellular viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). The MTT assay indicated high cell viability; maximum viability was obtained with the solution form of chitosan at day 5. Chitosan exposure increased the number of intercellular junctions and showed a significant proliferative effect on 3T3 mouse fibroblasts.     

Indole-based hydrazide-hydrazones and 4-thiazolidinones: synthesis and evaluation as antitubercular and anticancer agents

A new series of indolylhydrazones (6) and indole-based 4-thiazolidinones (7, 8) have been designed, synthesized and screened for in vitro antitubercular activity against Mycobacterium tuberculosis H37Rv. 4-Thiazolidinone derivatives 7g–7j, 8g, 8h and 8j displayed notable antituberculosis (anti-TB) activity showing 99% inhibition at MIC values ranging from 6.25 to 25.0?µg/ml. Compounds 7g, 7h, 7i, 8h and 8j demonstrated anti-TB activity at concentrations 10-fold lower than those cytotoxic for the mammalian cell lines. The indolylhydrazone derivative 6b has also been evaluated for antiproliferative activity against human cancer cell lines at the National Cancer Institute (USA). Compound 6b showed an interesting anticancer profile against different human tumor-derived cell lines at sub-micromolar concentrations with obvious selectivity toward colon cancer cell line COLO 205.     

Assignment of biological functions to specific plasmids in a local isolate ofRhizobium cicer

The genetic basis of symbiosis, mucoid structure and melanin biosynthesis in a local isolate ofRhizobium cicer was investigated. The strain was a very effective symbiont and produced substantial amounts of exopolysaccharides and melanin. It harbored three high-molar-mass plasmids of 55, 80 and 130 kb, respectively. Thirty-one melanin production-negative (Mep⁻) derivatives were obtained through plasmid curing. The plasmid patterns of cured derivatives indicated that the loss of 55 and/or 80 kb plasmids was sufficient for obtaining Mep⁻ phenotype. The specific involvement of 55-kb plasmid in melanin production was confirmed upon re-introduction of this plasmid into one of the Mep⁻ derivatives. Further investigation also indicated that the 130-kb plasmid might be necessary for both mucoidal appearance and symbiotic functions.     

Biodiversity of palm fungi in the tropics: are global fungal diversity estimates realistic?

Two questions are addressed: 'How many species of fungi can occur on a single host palm?' and 'What are the implications of this for global estimates of fungal diversity?' Fungal diversity estimates found in the literature are reviewed. Data on the numbers of fungi occurring on the above-ground tissues of six individual palms in the genus Licuala in Australia and Brunei Darussalam (Borneo) are provided. A total of 189 species of fungi were isolated and/or collected from the six palms. In addition, 53 'morphospecies' of mycelia sterilia were isolated, giving a total of 242 taxa from the 2672 isolates/collections made. The three palms in Australia (sampled once) yielded 100 species (each palm supporting an average of 54.7 taxa), while the three palms in Brunei Darussalam (sampled three times) yielded 172 species in total (approximately 111.3 taxa each). The magnitude of global fungal diversity, estimated at 1.5 million species, is discussed. Our results indicate that 33 to 1 would be a more accurate estimate (than 5.7 to 1) of the ratio of host specific fungal to palm species in the tropics. We therefore propose that global estimates of fungal diversity, based on temperate studies, require revision upwards.     

Isolation and identification of alkaline protease producer halotolerantBacillus licheniformis strain BA17

An alkaline protease producerBacillus licheniformis strain was isolated from Van Lake in Turkey. The strain is Gram positive, aerobic, motile, sporulating rod-shaped bacterium. Spores were ellipsoidal and positioned central in nonswollen sporangium. The cells were able to grow well at a pH range of 5.7–10. The optimal growth temperature was found to be 37 °C. Growth at a wide range of NaCl concentration (from 0 to 20%) showed that BA17 is halotolerant. Main fatty acid composition of BA17 was anteiso-C15:0 and iso-C15∶0. The strain was presumptively identified asB. licheniformis according to 16S rDNA gene sequence analysis. The most appropriate medium for the growth and protease production is composed of 0.5% yeast extract, 0.5% NaNO₃, 0.02% MgSO₄\7H₂O, 0.1% K₂HPO₄ and 0.5% maltose. The optimum temperature and pH of the alkaline protease of strain BA17 were found to be 60 °C and pH 11, respectively. The activity was completely lost in the presence of PMSF, suggesting that the preparation contains serine-alkaline protease(s).     

Screening Radiolocation Datasets for Movement Strategies With Time Series Segmentation

ABSTRACT Classical home range analysis is tailored to meet requirements of data with few points per individual with relatively large intervals between observations. The swift rise in Global Positioning System (GPS)-based studies requires the development of new analytical approaches because GPS data allow for more detailed analysis in time and space. The amount of data derived from GPS studies enhances the potential to more accurately separate movement strategies. We present a general, simple, conceptual approach to using large movement datasets to automatically screen and delimit spatial and temporal home ranges of individuals and movement strategies using time series segmentation. We used GPS data for moose (Alces alces) from a boreal Swedish population as an example. We tested predictions that our screening method could separate seasonal migration from dispersal and nomadic strategies by the movement profile, which includes several dimensions. Our analysis showed that broad strategies were detected using our simple analytical approach, which speeds up use of GPS data for management and research because the method can be used to calculate more objective spatial and temporal activity ranges in relation to movement strategies. Our examples illustrate the importance of using the time stamp on location data in describing home ranges and movements.     

Nisin production in a chitin-including continuous fermentation system with Lactococcus lactis displaying a cell wall chitin-binding domain

The limiting factors in the continuous production of nisin are high amount of biomass loss and low dilution rate application. In this study, a chitin-including continuous nisin fermentation system (CICON-FER) was constructed for high volumetric nisin production using nisin producer L. lactis displaying cell wall chitin-binding domain (ChBD) together with chitin in the reactor. In this respect, the highest binding conditions of relevant L. lactis cells to chitin were determined. Then the chitin flakes carrying nisin-producing L. lactis cells were used within the CICON-FER system at different dilution rates (0.1–0.9 h^?1) and initial glucose concentrations (20–60 g l^?1). The results revealed that the pH 7 conditions and the use of 100 mM sodium phosphate buffer with 0.1 % Tween 20 and Triton X-100 significantly increased the binding capacity of ChBD displaying L. lactis cells to chitin. The constructed CICON-FER system maintained the presence of the ChBD surface displaying L. lactis cells in the reactor system until 0.9 h^?1 dilution rate that resulted in a considerably high level of volumetric nisin production and productivity (10,500 IU ml^?1 and 9,450 IU ml^?1 h^?1, respectively) with the combination of a 0.9-h^?1 dilution rate and a 40-g l^?1 initial glucose concentration. In conclusion, an innovative nisin fermentation system that yielded the highest nisin production thus far and that was feasible for industrial application was created.     

Analysis of the fruit surfaces in Bupleurum L. (Umbelliferae) with SEM

20 Bupleurum L. taxa were investigated for micromorphological typology of the fruit surfaces with SEM. Considerably different types of sculpturing at species and subspecies level and similarities between related taxa were observed. A dichotomic key was prepared using the diagnostic surface patterns. Reticulate (sect. Bupleurum), rugose (sect. Aristata Godron subsect. Aristata) and rugulose-colliculate-verrucate (subsect. Juncea Briq.) types of surface structures were distinguished. Micromorphological characteristics may be valuable additional taxonomical criteria for the delimitation of infrageneric groups of species.