Gene expression during 3T3-L1 adipocyte differentiation. Characterization of initial responses to the inducing agents and changes during commitment to differentiation.

The mouse 3T3-L1 fibroblastic cell line rapidly differentiates to an adipocyte phenotype when post-confluent cells are treated for 48 h in fetal calf serum-containing medium supplemented with 1 microM dexamethasone (D), 0.5 mM methylisobutylxanthine (M) and 10 micrograms/ml insulin (I). D and I act synergistically to commit the cells to differentiate 24-48 h after initiating treatment, and this is blocked by the phorbol ester, 12-O-tetradecanoylphorbol-13-acetate. In order to identify cellular proteins involved in the differentiation process we analyzed differentiating 3T3-L1 cells using two-dimensional electrophoresis on large format gels. We observed changes in over 300 proteins during differentiation (over 100 within 5 h of initiating differentiation) and many of these are also changed at the level of mRNA (by analysis of in vitro translation products). About 75% of the initial changes were maximally induced by treatment with a combination of M and I, while no more than 10 proteins and their corresponding mRNAs were maximally induced by D within 3.5 h. Another 10 proteins were synergistically regulated by the combination of all three agents (DMI) within 3.5 h. Additional species were induced at later times. Five of these were synergistically induced by treatments that lead to differentiation, were first expressed at elevated levels during commitment and remained elevated in fully differentiated adipocytes. One or more of these proteins could well have a functional role in the commitment to and/or expression of the adipocyte differentiation program.     

Breast reconstruction in women treated with radiation therapy for breast cancer: cosmesis, complications, and tumor control.

The records of 55 patients who had breast cancer treated by mastectomy, irradiation, and breast reconstruction were reviewed for cosmetic outcome, complications, and tumor control. Median follow-up was 35 months. Local control rates were 95 percent in patients treated for high risk factors or breast conservation and 85 percent in patients treated for recurrent breast cancer. Acceptable cosmetic results were obtained in only 42 percent of patients. The incidence of complications was 55 percent. Transverse rectus abdominis muscle (TRAM) reconstructions gave superior cosmetic results compared with all other types of reconstructions. The timing of reconstruction in relation to mastectomy or radiation therapy did not significantly influence cosmetic outcome, although other factors suggest that delayed reconstruction may give better results. A majority of patients were satisfied with cosmetic outcome.     

Biodegradation of toxic and environmental pollutants.

Organic chemicals that are toxic to humans and to the environment can be transformed and metabolized by a variety of microorganisms. Such chemicals include trichloroethylene, chloroform, carbon tetrachloride, toluene, phenols, chlorinated phenols, polychlorinated biphenyls and polyaromatic hydrocarbons. This review focuses on some of the most important recent developments in the biodegradation of these toxic chemicals. Depending on the compound and the organism, the extent of our understanding ranges from the molecular level to the conceptual.     

Processing of precursor interleukin 1 beta and inflammatory disease

The processing of precursor interleukin 1 beta (IL1 beta) by elastase, cathepsin G, and collagenase, the major proteases released at sites of inflammation, was investigated using recombinant pro-IL1 beta. Each of these proteases cleaved the 31-kDa inactive precursor to a form similar in size and specific activity (greater than 10(8) units/mg) to the 17-kDa mature protein isolated from activated monocytes. Elastase, collagenase, and cathepsin G cleaved the IL1 beta precursor at distinct sites which are amino-terminal to the monocyte-processing site, Ala-117 (Cameron, P., Lumjuco, G., Rodkey, J., Bennett, C., and Schmidt, J. A. (1985) J. Exp. Med. 162, 790-801). Amino-terminal sequencing of the products of digestion by elastase and cathepsin G determined that resultant active IL1 beta proteins contained an additional 13 or 3 amino acids relative to mature IL1 beta. Synovial fluid collected from patients with inflammatory polyarthritis and bronchoalveolar lavage fluid from patients with sarcoidosis supplied similar processing activity(s). Control fluids from patients who had no symptoms of inflammatory disease did not exhibit processing activity. Lavage fluids that processed precursor IL1 beta were demonstrated to contain cathepsin G and/or elastase activity, whereas controls were negative. Because a significant fraction of IL1 beta may be secreted from monocytes as the inactive 31-kDa precursor (Hazuda, D. J., Lee, J. C., and Young, P. R. (1988) J. Biol. Chem. 263, 8473-8479, Bomford, R., Absull, E., Hughes-Jenkins, C., Simpkin, D., and Schmidt, J. (1987) Immunology 62, 543-549, and Mizel, S. B. (1988) in Cellular and Molecular Aspects of Inflammation Poste, G., and Crooke, S., eds) pp. 75-93, Plenum Publishing Corp., New York), these results suggest that in vivo the IL1 beta precursor can be processed after secretion by any of several proteases released at inflammatory sites.     

Attraction of the grasshopper, Melanoplus sanguinipes, to host plant odors and volatile components

The attraction of the polyphagous grasshopper, Melanoplus sanguinipes (F.), to odors from plant foliage or to chemical components of these odors was tested in a glass Y-tube olfactometer. Humidified air was passed at equal flow rates through a sample chamber containing plant material and an empty control chamber and then through the Y-tube to the holding chamber containing the test insect. Solutions of volatile chemicals in water were metered at a constant rate into the sample air stream with a syringe pump. Insects moving upwind were recorded after entering either the sample or control arm of the Y-tube. Both nymphs and adults were strongly attracted to the odor of chopped and intact seedling foliage of perennial ryegrass and wheat. Chopped leaves but not intact leaves of sorghum and alfalfa also were significantly attractive. The major components of ryegrass odor from both chopped and stem-cut leaves were Z-hex-3-en-1-yl acetate, Z-hex-3-en-1-ol, and pent-1-en-3-ol, in that order, with lesser amounts of E-hex-2-enal. Chopped leaves released many more minor components. The major components when tested individually or in binary and ternary mixtures were significantly attractive to grasshoppers compared to humidified air but not to the degree of whole plant odor. However, a quartenary mixture simulating the odor blend had levels of attractancy equal to that of chopped grass odor. Blends of these 5- and 6-carbon unsaturated alcohols, esters, and aldehydes volatilizing from green plants probably play an important role as olfactory cues for orientation of grasshoppers to food plants.

---

Résumé L'attraction de M. sanguinipes (F.) par l'odeur du feuillage et des composés chimiques de ces odeurs a été examinée dans un olfactomètre en tube de verre en Y. Des flux identiques d'air humidifié étaient introduits dans une enceinte contenant le matériel végétal et dans une enceinte témoin, et, via le tube en Y, dans l'enceinte contenant l'insecte à étudier. Des solutions aqueuses de substances volatiles étaient introduites à un taux constant dans le courant d'air grâce à une seringue. Les insectes se déplaçant contre le vent étaient enregistrés après pénétration dans une branche déterminée du tube en Y. Larves et adultes étaient fortement attirés par l'odeur de feuillage intact ou coupé de semis de Lolium perenne et de blé (Triticum aestivum). Les feuilles coupées, mais non les feuilles intactes de sorgho (Sorghum bicolor) et de luzerne (Medicago sativa) étaient, elles aussi, significativement attractives. Les principales substances constituant de l'odeur de feuilles de Lolium perenne, tant coupées qu'intactes, étaient dans l'ordre de Z-hex-3-en-1-yl acétate, le Z-hex-3-en-1-ol et le pent-1-en-3-ol, et dans une moindre mesure, E-hex-2-enal. Les feuilles coupées libéraient beaucoup plus de composés secondaires. Les principaux constituants testés, seuls ou en mélanges binaires ou ternaires, étaient significativement attractifs, mais moins que l'odeur totale de la plante. Toutefois, un mélange quaternaire, simulant le mélange odorant, avait le même pouvoir attractif que l'odeur de la plante entière. L'union de ces alcools, esters et aldéhydes à 5- et 6-carbones non saturés, volatilisés à partir des plantes vertes, jouent probablement un grand rôle comme signaux orientateurs lors de l'attraction des criquets par leur plantes alimentaires.

     

Evidence for nonclassical alpha adrenoceptor blockade by prazosin in isolated rabbit blood vessels

In isolated rabbit ear artery, prazosin shifted the concentration-response curve for norepinephrine (NE) to the right and increased the slope of the curve in the ED₅₀ region. Prazosin caused a smaller shift to the right of the concentration-response curve for NE in rabbit small saphenous vein and had no effect on the slope in the ED₅₀ region. The prazosin-induced increase in the slope of the NE concentration-response curve did not occur in the ear artery when neuronal and extraneuronal uptake were inhibited by desipramine (DMI) and deoxycorticosterone acetate (DOCA). Prazosin concentration ratios were significantly larger in the ear artery than in the small saphenous vein at all prazosin concentrations in the presence or absence of DMI and DOCA. Significant differences were found between prazosin dissociation constants (K_B) determined at three concentrations of prazosin in ear artery. No such differences were found between phentolamine dissociation constants determined in this vessel. Prazosin dissociation constants determined in small saphenous vein were higher than those determined in ear artery. These findings suggest that prazosin behaves as a noncompetitive antagonist in ear artery and that there are differences between the alpha adrenoceptors of the ear artery and those of the small saphenous vein.