At the Desert's Green Edge: An Ethnobotany of the Gila River Pima.

At the Desert's Green Edge: An Ethnobotany of the Gila River Pima. Amadeo M. Rea. Tucson: University of Arizona Press, 1997. 430 pp.     

A monoclonal antibody recognizes a 65 kDa higher plant membrane polypeptide which undergoes cation-dependent association with callose synthase in vitro and co-localizes with sites of high callose deposition in vivo

Summary A monoclonal antibody (MAb) capable of immobilizing detergent-solubilized UDP-glucose: (13)--glucan (callose) synthase activity from higher plants has been selected and characterized. On Western blots this MAb recognizes a polypeptide of about 65 kDa found in membranes isolated from a variety of plant sources. The polypeptide recognized by this MAb does not appear to bind the substrate UDP-glucose, and evidence is presented which indicates that this polypeptide associates with the enzyme complex in a cation-dependent manner under conditions where the callose synthase assumes a larger size. Indirect immunofluorescence localization with this MAb was positive with sieve plates of cucumber (Cucumis sativus) seedlings, and with plasmodesmata of onion (Allium cepa) epidermal cells, both being sites of localized, stress-induced callose deposition.Abbreviations BSA bovine serum albumine - DMSO dimethylsulf-oxide - DTT dithiothreitol - FITC fluorescein isothiocyanate - HB Hepes buffer - HBS Hepes buffer plus 0.15 M NaCl - IgG immuno-globulin G - MAb monoclonal antibody - MSB microtubule stabilizing buffer - NP 40 Nonidet P 40 - PBS phosphate buffered saline - SDS-PAGE sodium dodecylsulfate polyacrylamide gel electrophoresis - UDP uridine diphosphate - UV ultraviolet     

Plattenanomalien beiEchinocorys sulcata (Goldfuss 1826) aus dem Dan (Alttertiär) von Dalbyover,Dänemark

In a collection of 2000 specimens of the echinoidEchinocorys sulcata from the Danian of Dalbyover, northern Jutland, Denmark, two percent of the coronae investigated show characteristic coronal plate anomalies which are largely missing in other echinoid species. These anomalies are formed by split coronal plates or mosaic-like plate arrangements instead of regularly grown plates. Plate anomalies occurring in ambulacral and interambulacral zones, classified into 14 different types, have been used for a statistical analysis. Data on the relative age of the echinoid animals with anormal coronal plates, and on the most abundant types of plate anomalies are given. Two specimens ofEchinocorys sulcata from the same locality show deviations from the regular pentameric pattern and comparable finds from other localities and of otherEchinocorys species were incorporated in the analysis. The anomalies described here are named forma aegrainconstans n.f. Literature data on echinoid coronal plate anomalies, which have been related to coronal deformations caused by evironmental parameters, are discussed.     

Protein tracking and detection of protein motion using atomic force microscopy.

Height fluctuations over three different proteins, immunoglobulin G, urease, and microtubules, have been measured using an atomic force microscope (AFM) operating in fluid tapping mode. This was achieved by using a protein-tracking system, where the AFM tip was periodically repositioned above a single protein molecule (or structure) as thermal drifting occurred. Height (z-piezo signal) data were taken in 1 - or 2-s time slices with the tip over the molecule and compared to data taken on the support. The measured fluctuations were consistently higher when the tip was positioned over the protein, as opposed to the support the protein was adsorbed on. Similar measurements over patches of an amphiphile, where the noise was identical to that on the support, suggest that the noise increase is due to some intrinsic property of proteins and is not a result of different tip-sample interactions over soft samples. The orientation of the adsorbed proteins in these preliminary studies was not known; thus it was not possible to make correlations between the observed motion and specific protein structure or protein function beyond noting that the observed height fluctuations were greater for an antibody (anti-bovine IgG) and an enzyme (urease) than for microtubules.     

Interspecific and temporal variation in herbivore responses to hybrid willows

We studied herbivory of two species of willows (Salix sericea and S. eriocephala) and their interspecific hybrids to test alternative hypotheses concerning the effects of hybridization on plant resistance. Individually marked plants were identified using morphological traits in the field and random amplified polymorphic DNA (RAPD) band analysis was used to verify the genetic status of many parental and hybrid plants. The desities of 12 herbivore species on plants in the field were compared between two parents and their F₂-type hybrids. We found about equal support for the additive, dominance, and hybrid susceptibility hypotheses over 4 years. In one year, one species supported the hybrid resistance hypothesis. Guild membership was not a good predictor of similar responses of species to hybrid versus parental plants. There were marked differences in support for particular hypotheses among years for four herbivore species. This study demonstrates the diversity of responses of phytophages in response to interspecific hybridization, and indicates that year-to-year variation in relative resistance of hybrid plants can be important.     

Protease inhibitor localization in control and streptozotocin-diabetic skeletal muscles

Summary ₁-Antitrypsin and ₁-inhibitor-3 were localized for the first time inside skeletal muscle cells. Their content, especially that of ₁-inhibitor-3, was greatly reduced following streptozotocin-induced diabetes. ₁-Antitrypsin and ₁-inhibitor-3 were also observed in the vascular components and interstitial space surrounding both control and diabetic soleus muscles as revealed by immunofluorescence. In diabetic muscles, the non-myofibre locale of ₁-inhibitor-3 was reduced, and to a lesser extent, ₁-antitrypsin. Both myofibre and extracellular patterns were reversed to control levels by insulin replacement.     

Purification to homogeneity, characterization and monoclonal antibodies of phospholipid-sensitive Ca2+-dependent protein kinase from spleen.

A phospholipid-sensitive Ca2+-dependent protein kinase was purified to homogeneity, for the first time, from extracts of pig spleen, employing the steps of DEAE-cellulose, octyl-agarose, Sephacryl S-200 and phosphatidylserine-Affigel 10 affinity chromatographies. The purified enzyme appeared as a single protein band on both analytical (non-denaturing) and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, having a minimum mol.wt. of 68 000 +/- 200. The molecular weight of the enzyme was also determined to be 74 500 +/- 4600 by gel filtration and 80 000 based on its sedimentation coefficient (5.52 S) and Stokes radius (3.52 +/- 0.09 nm), indicating that the enzyme was a monomeric protein. The frictional ratio (f/f0) of the enzyme was 1.24, indicating it was non-globular in shape. The enzyme had a pI of 5.3, and a pH optimum of 6.5 for its reaction. Amino acid analysis indicated that the enzyme apparently was not similar to myosin light-chain kinase (a calmodulin-sensitive species of Ca2+-dependent protein kinase) or cyclic AMP-dependent and cyclic GMP-dependent protein kinases. The enzyme had an apparent Km for ATP of 7.5 microns. Histone H1 and myelin basic protein were effective substrates for the enzyme, with apparent Km values of 0.3 and 0.2 microns, and Vmax, values of 0.06 and 0.09 mumol/min per mg of enzyme respectively. The enzyme activity was dependent on both phosphatidylserine (apparent Ka = 6.25 micrograms/ml) and Ca2+ (apparent Ka = 160 microns). Calmodulin was unable to substitute for the phospholipid as a cofactor, nor was it a subunit of the enzyme. Sr2+ and Ba2+ could partially mimic Ca2+ to activate the enzyme in the presence of phosphatidylserine. An endogenous substrate protein (mol.wt. 41 000) for the enzyme was found in the total, solubilized fraction of pig spleen. Monoclonal antibodies against the enzyme interacted similarly with the homogeneous and impure enzyme; the antibodies, however, did not bind to cyclic nucleotide-dependent protein kinases.     

Evidence for a role of the monoclonal antibody E48 defined antigen in cell-cell adhesion in squamous epithelia and head and neck squamous cell carcinoma

Monoclonal antibody (MAb) E48 recognizes a 20- to 22-kDa antigen expressed by human squamous and transitional epithelia and their neoplastic counterparts. Histochemical examination of these tissues revealed distinct surface labeling with MAb E48. To investigate the subcellular localization of the E48 antigen we have performed electron microscopical analysis. In cells of normal oral mucosa, the E48 antigen was expressed on the plasmalemma, particularly associated with desmosomes, suggesting involvement of the E48 antigen in intercellular adhesion. Furthermore, the level of expression of the E48 antigen appeared to be influenced by the cellular organization. In squamous cell carcinoma (SCC) cell lines grown in vitro as subconfluent monolayer cultures, the E48 antigen expression was low. However, E48 antigen expression increased when SCC cells were grown to confluency. E48 antigen expression was similarly high when SCC cell lines were cultured under conditions promoting three-dimensional growth either as colonies within floating collagen gels or as xenograft in tumor-bearing nude mice. Further evidence for the involvement of the E48 antigen in cell-cell adhesion was found when SCC cells were grown within collagen gels in the presence of MAb E48: no spherical colonies were formed, but cells grew out to colonies composed of single cells. Moreover, in this culture system the percentage of SCC cells growing out to colonies was decreased by the presence of MAb E48. These findings indicate that the E48 antigen is involved in the structural organization of squamous tissue and might have a role in intercellular adhesion.