Foliar flavonoids of eastern North AmericanVitis (Vitaceae) north of Mexico

Twenty-one flavonoid glycosides were isolated from the leaves of 22 North AmericanVitis L. taxa, representing two subgenera and five series. Three chemical groups were evident: one producing flavonols, flavones, and C-glycosylflavones, a second producing flavonols and flavones, and a third producing only flavonols. These three chemical groups did not correspond to any of the subgeneric groupings based on morphology. However, flavonoid distributions within series in each subgenus correlate well with morphological data. Parallel flavonoid evolution within each series is thought to account for this lack of subgeneric and interserial flavonoid distinction. The flavonoid data indicate that seriesCordifoliae of subgenusVitis, particularlyV. vulpina L., is the most closely related group to subgenusMuscadinia (Planch.)Rehder, and represents an evolutionary link between the two subgenera.     

CHARACTERIZATION OF A BOTTLENOSE DOLPHIN (TURSIOPS TRUNCATUS) KIDNEY EPITHELIAL CELL LINE

Abstract: An epithelial cell line, Carvan dolphin kidney (CDK), isolated from a prematurely born female bottlenose dolphin, Tursiops truncatus , exhibited growth characteristics not previously reported for cetacean cells in culture. CDK cells were cytokeratin positive and demonstrated a maximum doubling time of 1.31 days, with plating and colony forming efficiencies approaching 100% for the early population doublings. Despite an unusually efficient colony-forming ability and rapid growth, these cells were neither transformed nor immortal, displaying normal contact inhibition, anchorage dependence, and the requirement for high concentrations of fetal bovine serum in the growth medium. CDK cells exhibited age-dependent changes in growth rate, colony-forming efficiency, and cytoplasmic profile, and showed a finite lifespan of about 50 population doublings and a stable 2N = 44 karyotype which correlates with previously reported cytogenetic analyses. Velocity sedimentation analysis showed that CDK cells contained nuclear aryl hydrocarbon receptor (Ah), indicating their potential to be induced for cytochrome P450. These data suggest that CDK cells may have utility as an in vitro toxicological model for evaluating hydrocarbon contaminant effects on Tursiops truncatus , a protected marine mammal.     

Isolation and characterization of a hemocyte aggregation inhibitor from hemolymph of Manduca sexta larvae

A protein that inhibits hemocyte aggregation has been isolated from hemolymph of Manduca sexta larvae and named hemocyte aggregation inhibitor protein (HAIP). HAIP has a M_r = 50,000, pI = 8.5, and contains 7% carbohydrate. It is present at 230 ± 20 μg/ml in hemolymph of day 3 fifth instar larvae. Antibodies to HAIP do not cross-react with M. sexta hemolin, which is similar in size and charge and also inhibits hemocyte aggregation. HAIP and hemolin have some similarity in amino acid composition and NH₂-terminal sequence, but are different in overall secondary structure, as determined by CD spectroscopy. The concentration of HAIP in hemolymph is not affected by injection of larvae with bacteria. A protein of approximately 50,000 daltons that reacts with antibody to M. sexta HAIP is present in hemolymph of Bombyx mori, Heliothis zea, and Galleria mellonella. Although the function of HAIP in vivo is not yet clear, it may have a role in modulating adhesion of hemocytes during defensive responses. © 1994 Wiley-Liss, Inc.     

A method for α-helical integral membrane protein fold prediction

Integral membrane proteins (of the α-helical class) are of central importance in a wide variety of vital cellular functions. Despite considerable effort on methods to predict the location of the helices, little attention has been directed toward developing an automatic method to pack the helices together. In principle, the prediction of membrane proteins should be easier than the prediction of globular proteins: there is only one type of secondary structure and all helices pack with a common alignment across the membrane. This allows all possible structures to be represented on a simple lattice and exhaustively enumerated. Prediction success lies not in generating many possible folds but in recognizing which corresponds to the native. Our evaluation of each fold is based on how well the exposed surface predicted from a multiple sequence alignment fits its allocated position. Just as exposure to solvent in globular proteins can be predicted from sequence variation, so exposure to lipid can be recognized by variable-hydrophobic (variphobic) positions. Application to both bacteriorhodopsin and the eukaryotic rhodopsin/opsin families revealed that the angular size of the lipid-exposed faces must be predicted accurately to allow selection of the correct fold. With the inherent uncertainties in helix prediction and parameter choice, this accuracy could not be guaranteed but the correct fold was typically found in the top six candidates. Our method provides the first completely automatic method that can proceed from a scan of the protein sequence databanks to a predicted three-dimensional structure with no intervention required from the investigator. Within the limited domain of the seven helix bundle proteins, a good chance can be given of selecting the correct structure. However, the limited number of sequences available with a corresponding known structure makes further characterization of the method difficult. © 1994 John Wiley & Sons, Inc.     

Crystallization and preliminary crystallographic studies of the precursor and mature forms of a neutral lipase from the fungus Rhizopus delemar

A neutral lipase from the filamentous fungus Rhizopus delemar has been crystallized in both its proenzyme and mature forms. Although the latter crystallizes readily and produces a variety of crystal forms, only one was found to be suitable for X-ray studies. It is monoclinic (C2, a = 92.8 Å, b = 128.9 Å, c = 78.3 Å, β = 135.8) with two molecules in the asymmetric unit related by a noncrystallographic diad. The prolipase crystals are orthorhombic (P2₁2₁2₁, with a = 79.8 Å, b = 115.2 Å, c = 73.0 Å) and also contain a pair of molecules in the asymmetric unit. Initial results of molecular replacement calculations using the refined coordinates of the related lipase from Rhizomucor miehei identified the correct orientations and positions of the protein molecules in the unit cells of crystals of both proenzyme and the mature form. © 1994 John Wiley & Sons, Inc.     

The molecular genetics of Alzheimer's disease

The major pathological characteristic of Alzheimer's disease (AD) is the abnormal deposition of β-amyloid peptide (Aβ) in the brain. In some early onset cases, the disease develops because of mutations in the gene coding for β-amyloid precursor protein (βAPP). However, the majority of AD families in the early onset subgroup are linked to a locus on chromosome 14. The genetic analysis and age of onset correlates of both the βAPP gene and the chromosome 14 locus are discussed. We speculate on the mechanisms by which the βAPP mutations cause the disease and discuss recent advances in βAPP processing that may be relevant to the pathogenesis of the late-onset (common) form of the disease. In addition, we review the association of theAPOE locus with late-onset familial and nonfamilial disease. Further work is required to establish the effects of this locus on disease occurrence, age of onset, and progression. The molecular pathology of ApoE in relation to AD development and the identification of the chromosome 14 gene will greatly contribute to a general pathogenic model of AD, and will clarify the role of βAPP and its derivatives.     

Seasonal growth activity of local and foreign gracilarioid strains in Israel

Six gracilarioid strains originating from different climatical environments were cultured in two cultivation systems: a short-term indoor one with a cross gradiant table, and a long-term outdoor one. Seasonal growth performances of the different strains were determined. The growth results in the two culture systems showed similar trends. The tropical species.Gracilaria cornea andG. cornea mutant, showed highest growth rates during summer and no growth at all during winter. The temperate species,Gracilaria verrucosa andGracilariopsis lemaneiformis, showed best growth performances during winter with small fluctuations between seasons. The subtropical speciesGracilaria conferta (local) showed seasonal growth fluctuations all over the year. The foreign species definitely did not acclimate under local conditions, but successfully preserved their original response to temperature. Regression equations confirmed that temperature was the dominant environmental variable in most of the gracilarioid strains. The growth rate results obtained showed encouraging prospects for high algal productivity as compared to other cultivation systems. Seasonal cultivation strategy ofGracilaria spp. in Israel is discussed.