Efficient regeneration of Brassica oleracea hypocotyl protoplasts and high frequency genetic transformation by direct DNA uptake

Summary Efficient regeneration (80%) and high frequency genetic transformation (10–33%) were achieved by culturing protoplasts isolated from hypocotyl tissues of six day old Brassica oleracea seedlings and by subjecting these protoplasts to PEG mediated direct plasmid uptake. Three different plasmid vectors carrying marker genes for resistance to methotrexate (dhfr), hygromycin (hpt) and phosphinotricin (bar) were constructed and used for transformation. Large number of normal, fertile transformants were obtained with vectors carrying hpt and bar genes. No transformants could be regenerated for resistance to methotrexate as it severely suppressed shoot differentiation.Abbreviations bar/PAT bialaphos resistance gene/phosphinotricin acetyltransferase - 2,4-D 2,4-di-chlorophenoxyacetic acid - dhfr/DHPR dihydrofolate reductase gene/enzyme - gus/GUS -glucuronidase gene/enzyme - hpt/HPT hygromycin phosphotransferase gene/enzyme - Kn kinetin - PEG polyethylene glycol - RH relative humidity     

Conservation biology: predicting birds' responses to forest fragmentation

Understanding species' ecological responses to habitat fragmentation is critical for biodiversity conservation, especially in tropical forests. A detailed recent study has shown that changes in the abundances of bird species following fragmentation may be dramatic and unpredictable.     

Somatic cell hybridization of ’oxy’ CMS Brassica juncea (AABB) with B. oleracea (CC) for correction of chlorosis and transfer of novel organelle combinations to allotetraploid brassicas

Alloplasmic lines of cultivated Brassica species with B. oxyrrhina cytoplasm are male-sterile and suffer from severe chlorosis. We developed male-sterile lines corrected for chlorosis by fusing protoplasts of CMS B. juncea (AABB) with ’oxy’ cytoplasm and normal B. oleracea (CC). A large number of male-sterile AABBCC somatic hybrids with desirable organelle combinations, i.e. chloroplasts of B. oleracea and mitochondria with recombinant genomes, were recovered. While no recombination was observed in the chloroplast genome, the mitochondrial genome showed extensive recombination that resulted in the appearance of totally novel banding patterns in some of the hybrids. Hybrids with a parental-type mitochondrial genome as well as recombinant patterns close to either of the parental types were also obtained. Using AABBCC somatic hybrids as bridging material, we transferred the desirable organelle combinations to B. juncea (AABB), B. napus (AACC), and B. carinata (BBCC). Many of these lines are now at advanced stages of backcrossing and show stable inheritance of the CMS character and do not suffer from chlorosis. Received: 9 August 1999 / Accepted: 14 September 1999     

Iodine-mediated one-pot intramolecular decarboxylation domino reaction for accessing functionalised 2-(1,3,4-oxadiazol-2-yl)anilines with carbonic anhydrase inhibitory action

A practical and transition metal-free one-pot domino synthesis of diversified (1,3,4-oxadiazol-2-yl)anilines has been developed employing isatins and hydrazides as the starting materials, in the presence of molecular iodine. The prominent feature of this domino process involves consecutive condensation, hydrolytic ring cleavage, and an intramolecular decarboxylation, in a one-pot process that leads to the oxidative formation of a C–O bond. Fluorescence properties of some of the representative molecules obtained in this way were studied. The synthesised 2-(1,3,4-oxadiazolo-2-yl)aniline-benzene sulphonamides (8a–o) were screened for their carbonic anhydrase (CA, EC 4.2.1.1) inhibitory activity. Most of the compounds exhibited low micromolar to nanomolar activity against human (h) isoforms hCA I, hCA II, hCA IV, and XII, with some compounds displaying selective CA inhibitory activity towards hCA II with K_Is of 6.4–17.6?nM.     

Effect of cisplatin, lipopolysaccharide, muramyl dipeptide and recombinant interferon-gamma on murine macrophages in vitro. I. Macrophage-mediated tumor cell lysis

Murine macrophage monolayers treated with cisplatin, lipopolysaccharide (LPS), muramyl dipeptide (MDP) or recombinant interferon-gamma (rIFN gamma) were observed to have significantly increased tumoricidal activity. rIFN gamma had synergistic effects with cisplatin, LPS or MDP in activating macrophages. However, MDP showed much more pronounced synergism with cisplatin and LPS than with rIFN gamma. Supernatants collected from these activated macrophage monolayers also showed increased tumoricidal activity. Tumor cell lysis mediated by cisplatin-treated macrophages did not require priming with rIFN gamma though it may be necessary as a first signal for the increased macrophage activation with LPS and MDP.     

Down's syndrome and cardiac tamponade with pulmonary tuberculosis in adults

We describe a combination case of Down''s syndrome and Cardiac Tamponade with Pulmonary Tuberculosis in an adult patient.     

Relative need for conservation assessments of vascular plant species among ecoregions

Aim (1) To determine the relative need for conservation assessments of vascular plant species among the world’s ecoregions given under‐assessed species distributions; (2) to evaluate the challenge posed by the lack of financial resources on species assessment efforts; and (3) to demonstrate the utility of nonlinear mixed‐effects models with both homoscedastic and heteroscedastic error structures in the identification of species‐rich ecoregions. Location Global. Methods We identified the world’s ecoregions that contain the highest vascular plant species richness after controlling for area using species–area relationship (SAR) models built within a mixed‐effects multi‐model framework. Using quantitative thresholds, ecoregions with the highest plant species richness, historical habitat loss and projected increase in human population density were deemed to be most in need of conservation assessments of plant species. We used generalized linear models to test if countries that overlap with highly important ecoregions are poorer compared with others. Results We classed ecoregions into nine categories based on the relative need for conservation assessments of vascular plant species. Ecoregions of highest relative need are found mostly in the tropics, particularly Southeast Asia, Central America, Tropical Andes and the Cerrado of South America, and the East African montane region and its surrounding areas. Countries overlapping with ecoregions deemed important for conservation assessments are poorer as measured by their capita gross national income than the other countries. The nonlinear mixed modelling framework was effective in reducing residual spatial autocorrelation compared with nonlinear models comprised of only fixed effects. In contrasting multiple SAR models to identify species‐rich ecoregions, there was not one SAR model that fitted best across all biomes. Not all SAR models displayed homoscedastic errors; therefore it is important to consider models with both homoscedastic and heteroscedastic error structures. Main conclusions We propose that conservation assessments should be conducted first in ecoregions with the greatest predicted species richness, historical habitat loss and future human population increase. As ecoregions deemed to be important for conservation assessments are located in the poorest countries, we urge international aid agencies and botanic gardens to cooperate with both local and international scientists to fund and implement conservation assessment programmes there.