Role of TAP-1 and/or TAP-2 antigen presentation defects in tumorigenicity of mouse melanoma

Mutations in transporters associated with antigen processing (TAP-1 and -2) required for the transport of cytosolic endogenous peptides to the endoplasmic reticulum correlate with increased metastatic potential and reduced host survival in several malignancies. To address the possible function of TAP as a "tumor suppressor" gene, we show that correction of TAP-1 and/or TAP-2 defects in B16 mouse melanoma enhanced the cell surface expression of MHC class I molecules and significantly reduced the rate of subcutaneous tumor growth and pulmonary metastatic burden. Cytotoxic assays confirmed increased sensitivity of TAP-1 and/or TAP-2 transfected clones of B16 melanoma to cytotoxic T lymphocytes. These results indicate that the expression of TAP limits the malignant potential of tumors with implications for CD8(+) T cell-based immunotherapy in controlling growth of certain TAP-deficient malignancies.     

Vegetative survival and reproduction under submerged and air-exposed conditions and vegetative survival as affected by salts,pesticides, and metals in aerial green alga<Emphasis Type="Italic">Trentepohlia aurea</Emphasis>

Trentepohlia aurea vegetative cells do not survive submerged conditions for more than 5 months, but can survive air-exposed conditions for more than 1 year. Disintegration and rapid death of algal cells was observed to a higher extent under submerged than air-exposed condition. Under submerged conditionsT. aurea did not form any sporangium while prolific formation occurred under air-exposed conditions. Under submerged conditions algal cells formed few-celled, filamentous, cytoplasmic type setae. Vegetative cells were resistant to some extent to various levels of salt (NaCl, ≤0.8 mol/L), pesticides (DDT, 2,4-D or captan, 2000 ppm) and ‘heavy’ metals (zinc or nickel, 200 ppm; cobalt, ≤100 ppm.)     

Polymorphic buttonwood: effects of disturbance on resistance to herbivores in green and silver morphs of a Bahamian shrub

We studied consequences of storm damage on buttonwood (Conocarpus erectus) shrubs and their herbivores in the Bahamian islands. Buttonwood is polymorphic, with green shrubs producing few leaf trichomes and silver shrubs covered in dense trichomes. We first characterize traits of green vs. silver shrubs relevant for herbivores, and then assay damage by two prominent insects. Next, on replicated islands, we experimentally address how different types of storm damage (simulated hurricane surge damage vs. simulated intense wind) affected phenotypic traits of both buttonwood morphs and subsequent herbivory over a one-year sampling period. Our results show that although leaves produced by green shrubs are 21% tougher than leaves produced by silver shrubs, green leaves have 16% higher nitrogen concentrations and greater levels of herbivory. Consistent with previous observational studies of a natural hurricane at our study site, we found stronger effects of simulated surge damage than simulated wind damage. Experimental pruning of shrubs resulted in reduced toughness, higher nitrogen concentration, fewer trichomes, and greater herbivory compared to controls and compared to shrubs with their leaves stripped. The results were stronger for the silver compared to the green morph. Morph differences in buttonwood have strong consequences for herbivores, and these effects are modified by disturbance.     

The bridging function of hepatic lipase clears plasma cholesterol in LDL receptor-deficient "apoB-48-only" and "apoB-100-only" mice

Hepatic lipase clears plasma cholesterol by lipolytic and nonlipolytic processing of lipoproteins. We hypothesized that the nonlipolytic processing (known as the bridging function) clears cholesterol by removing apoB-48- and apoB-100-containing lipoproteins by whole particle uptake. To test our hypotheses, we expressed catalytically inactive human HL (ciHL) in LDL receptor deficient "apoB-48-only" and "apoB-100-only" mice. Expression of ciHL in "apoB-48-only" mice reduced cholesterol by reducing LDL-C (by 54%, 46 +/- 6 vs. 19 +/- 8 mg/dl, P < 0.001). ApoB-48 was similarly reduced (by 60%). The similar reductions in LDL-C and apoB-48 indicate cholesterol removal by whole particle uptake. Expression of ciHL in "apoB-100-only" mice reduced cholesterol by reducing IDL-C (by 37%, 61 +/- 19 vs. 38 +/- 12 mg/dl, P < 0.003). Apo-B100 was also reduced (by 27%). The contribution of nutritional influences was examined with a high-fat diet challenge in the "apoB-100-only" background. On the high fat diet, ciHL reduced IDL-C (by 30%, 355 +/- 72 vs. 257 +/- 64 mg/dl, P < 0.04) but did not reduce apoB-100. The reduction in IDL-C in excess of apoB-100 suggests removal either by selective cholesteryl ester uptake, or by selective removal of larger, cholesteryl ester-enriched particles. Our results demonstrate that the bridging function removes apoB-48- and apoB-100-containing lipoproteins by whole particle uptake and other mechanisms.     

Large scale identification and quantitative profiling of phosphoproteins expressed during seed filling in oilseed rape

Seed filling is a dynamic, temporally regulated phase of seed development that determines the composition of storage reserves in mature seeds. Although the metabolic pathways responsible for storage reserve synthesis such as carbohydrates, oils, and proteins are known, little is known about their regulation. Protein phosphorylation is a ubiquitous form of regulation that influences many aspects of dynamic cellular behavior in plant biology. Here a systematic study has been conducted on five sequential stages (2, 3, 4, 5, and 6 weeks after flowering) of seed development in oilseed rape (Brassica napus L. Reston) to survey the presence and dynamics of phosphoproteins. High resolution two-dimensional gel electrophoresis in combination with a phosphoprotein-specific Pro-Q Diamond phosphoprotein fluorescence stain revealed approximately 300 phosphoprotein spots. Of these, quantitative expression profiles for 234 high quality spots were established, and hierarchical cluster analyses revealed the occurrence of six principal expression trends during seed filling. The identity of 103 spots was determined using LC-MS/MS. The identified spots represented 70 non-redundant phosphoproteins belonging to 10 major functional categories including energy, metabolism, protein destination, and signal transduction. Furthermore phosphorylation within 16 non-redundant phosphoproteins was verified by mapping the phosphorylation sites by LC-MS/MS. Although one of these sites was postulated previously, the remaining sites have not yet been reported in plants. Phosphoprotein data were assembled into a web database. Together this study provides evidence for the presence of a large number of functionally diverse phosphoproteins, including global regulatory factors like 14-3-3 proteins, within developing B. napus seed.     

Synthesis, biological evaluation and molecular docking of aryl hydrazines and hydrazides for anticancer activity

Aryl hydrazine and hydrazide analogues were synthesized based on p-tolyl hydrazine, isolated as a breakdown product of a secondary metabolite from the mushroom, Agaricus bisporus, and tested to be highly active molecule than 5-fluorouracil in in vitro anticancer studies. The synthesized analogues were tested for anticancer activity using NCI protocol. Anolgues 12 and 15 emerged as molecules with significant in vitro anticancer activity. Molecular docking study revealed the binding orientations of aryl hydrazines and hydrazides analogues in the active sites of thymidylate synthase.