Opposing effects on modulation of angiogenesis by protein kinase C and cAMP-mediated pathways

The role of cAMP-mediated pathway in modulating angiogenesis was investigated. We have shown previously that activators of protein kinase C (PKC) caused a marked increase in angiogenesis, while the specific inhibitor of PKC, Ro 318220 suppressed angiogenesis. Here we show that forskolin, which activates adenylate cyclase and elevates the intracellular levels of cAMP, and the Sp-diastereomer of adenosine cyclic-3',5'-monophosphothioate (Sp-cAMPS), caused a dose-dependent suppression of collagenous protein biosynthesis and angiogenesis in the chick chorioallantoic membrane system (CAM). The opposite modulation of angiogenesis by activators of PKC and elevated cAMP levels was further confirmed by the suppression of 4 beta-phorbol-12-myristate-13-acetate (4 beta-PMA)-stimulated angiogenesis by either forskolin or Sp-cAMPS. On the contrary, the Rp-diastereomer of adenosine cyclic-3',5'-monophosphothioate (Rp-cAMPS), which antagonises endogenous cAMP biochemical actions, had no effect on angiogenesis alone and did not suppress 4 beta-PMA stimulated angiogenesis. However, Rp-cAMPS antagonised the effect of forskolin and Sp-cAMPS on 4 beta-PMA induced angiogenesis. Similar results were obtained in the human umbilical vein endothelial cell tube formation assay. In this system, the PKC inhibitor, Ro 318220, caused a dose-dependent inhibition of 4 beta-PMA reversed this effect. Also, forskolin and Sp-cAMPS caused an inhibition in tube formation. These results indicate that increased levels of intracellular cAMP have a negative effect in normal angiogenesis and cause a large reduction of the promotion of angiogenesis resulting from PKC activation.     

Photoreceptor loss in age-related macular degeneration

PURPOSE: The authors showed previously that parafoveal rods, but not cones, decrease during the course of adulthood in donor eyes that were screened to exclude the grossly visible macular drusen and pigmentary disturbances typical of age-related macular degeneration (AMD). Because AMD begins in the parafovea, this selective loss of rods actually may be subclinical AMD not yet visible in the fundus. If so, AMD must have a predilection for rods over cones. The authors tested this hypothesis by determining the relative numbers of cones and rods in donor eyes with mid-to late-stage AMD and in age-matched controls. METHODS: Thirteen eyes (from seven donors) with grossly visible macular drusen and pigmentary disturbances were either wholemounted for photoreceptor counts or sectioned through the fovea for histopathology and carbonic anhydrase histochemistry to label red-green cones. Eyes were assigned to AMD or control groups on the basis of histopathology and clinical history. RESULTS: Five nonexudative AMD (NE-AMD) eyes from three donors showed sparing of foveal cones and loss of rods and cones in the parafovea. In two donors, rod loss exceeded cone loss at most parafoveal locations, and in one donor, rod density was normal and cone density was reduced. In eight exudative AMD (EX-AMD) eyes from five donors, photoreceptors surviving along the margins of and overlying disciform scars were largely cones. CONCLUSIONS: Photoreceptors are lost in NE-AMD as well as in the more severe exudative form, consistent with functional and clinical studies. The authors propose that rods die in older eyes without evidence of overt retinal pigment epithelial disease. In persons susceptible to AMD, the retinal pigment epithelium becomes dysfunctional. Secondarily, rod loss continues and cones begin to degenerate. Eventually, only degenerate cones remain; ultimately, all photoreceptors may disappear.     

Management of a pool of self-service cars

This paper deals with the problem of self-service cars arising in large companies where cars are at the employees' disposal to travel between different company sites. This problem also occurs in some big cities where electric cars are available for customers to make local journeys. The employee (or the customer) uses the car to travel from one site to another, and leaves it there so that other employees (or customers) can use it. This customer will use another car to travel to another site, if necessary. In this paper, we restrict ourselves to the case where the random variables which generate car requirements in a given site are independent identically distributed (i.i.d.) random variables, and we study the behaviour of the system according to a balancing strategy.     

Minor groove-directed and intercalative ligand-DNA interactions in the poisoning of human DNA topoisomerase I by protoberberine analogs

Spectroscopic, calorimetric, DNA cleavage, electrophoretic, and computer modeling techniques have been employed to characterize the DNA binding and topoisomerase poisoning properties of three protoberberine analogs, 8-desmethylcoralyne (DMC), 5,6-dihydro-8-desmethylcoralyne (DHDMC), and palmatine, which differ in the chemical structures of their B- and/or D-rings. DNA topoisomerase-mediated cleavage assays revealed that these compounds were unable to poison mammalian type II topoisomerase. By contrast, the three protoberberine analogs poisoned human topoisomerase I according to the following hierarchy: DHDMC > DMC > palmatine. DNA binding by all three protoberberine analogs induced negative flow linear dichroism signals as well as unwinding of the host duplex. These two observations are consistent with an intercalative mode of protoberberine binding to duplex DNA. However, a comparison of the DNA binding properties for DMC and DHDMC, which differ only by the state of saturation at the 5,6 positions of the B-ring, revealed that the protoberberine analogs do not "behave" like classic DNA intercalators. Specifically, saturation of the 5-6 double bond in the B-ring of DMC, thereby converting it to the DHDMC molecule, was associated with enhanced DNA unwinding as well as a reversal of DNA binding preference from a DNA duplex with an inaccessible or occluded minor groove {poly[d(G-C)]2} to DNA duplexes with accessible or unobstructed minor grooves {poly[d(A-T)]2 and poly[d(I-C)]2}. In addition, a comparison of the DNA binding properties for DHDMC and palmatine revealed that transferring the 11-methoxy moiety on the D-ring of DHDMC to the 9 position, thereby converting it to palmatine, was associated with a reduction in binding affinity for both duplexes with unobstructed minor grooves as well as for duplexes with occluded minor grooves. These DNA binding properties are consistent with a "mixed-mode" DNA binding model for protoberberines in which a portion of the ligand molecule intercalates into the double helix, while the nonintercalated portion of the ligand molecule protrudes into the minor groove of the host duplex, where it is thereby available for interactions with atoms lining the floor and/or walls of the minor groove. Furthermore, saturation at the 5,6 positions of the B-ring, which causes the A-ring to be tilted relative to the plane formed by rings C and D, appears to stabilize the interaction between the host duplex and the minor groove-directed portion of the protoberberine ligand. Computer modeling studies on the DHDMC-poly[d(A-T)]2 complex suggest that this interaction may involve van der Waals contacts between the ligand A-ring and backbone sugar atoms lining the minor groove of the host duplex. The hierarchy of topoisomerase I poisoning noted above suggests that this minor groove-directed interaction may play an important role in topoisomerase I poisoning by protoberberine analogs. In the aggregate, our results presented here, coupled with the recent demonstration of topoisomerase I poisoning by minor groove-binding terbenzimidazoles [Sun, Q., Gatto, B., Yu, C., Liu, A. , Liu, L. F., & LaVoie, E. J. (1995) J. Med. Chem. 38, 3638-3644], suggest that minor groove-directed ligand-DNA interactions may be of general importance in the poisoning of topoisomerase I.     

Cultural/ethnicity issues in the classroom

Discussing sensitive racial and ethnicity issues in the classroom may predispose to heated confrontation if not a legal tangle between the professor and student(s). Students and faculty alike experience uncomfortable feelings in discussing or giving opinions are expressed concerning "isms" (e.g., sexism, socialism, paternalism, feminism, religiosity/spiritualism practices). To minimize the occurrence of this likely situation, implementing a reasoned, systematic mode of discourse is suggested. As a foundation for this approach, modern and contemporary ethical theories are reviewed, and Baldonado's Ethical Decision-Making Paradigm is used to illustrate choosing a morally accepted course of action (act). Implications of culturally sensitivity issues in the classroom, curriculum and nursing practice are presented.     

12-O-tetradecanoylphorbol-13-acetate induces clonal expansion of potentially malignant keratinocytes in a tissue model of early neoplastic progression

Tumor promoters stimulate the selective expansion of initiated mouse keratinocytes in the two-stage model of skin carcinogenesis. However, it is not clear whether these promoters directly modulate the growth of initiated cells or rather permit clonal expansion of initiated cells by modifying the environment of adjacent normal cells. The goal of this study was to further understand the mechanism of action of tumor promotion during early neoplastic progression of human stratified epithelium. To accomplish this, we have established an organotypic culture model that mimics a preneoplastic tissue and contains mixtures of genetically marked (beta-galactosidase), low-grade malignant keratinocytes (HaCaT-ras II-4) and normal human keratinocytes (NHKs) to monitor the fate and phenotype of these cells after treatment with 12-O-tetradecanoylphorbol-13-acetate (TPA). In submerged culture, concentrations of 0.001-1 microg/ml TPA were shown to limit the growth of NHKs yet had no effect on growth of II-4 cells. TPA (0.001 microg/ml) was then added to organotypic cultures containing mixtures of NHK:II-4 cells at varying ratios to determine whether this agent could selectively stimulate clonal expansion of II-4 cells in a normal epidermal background. Immunofluorescence for beta-galactosidase demonstrated that TPA caused a significant increase in the percentage of beta-galactosidase-positive areas in 12:1 and 4:1 mixtures. This TPA-induced expansion of II-4 cells was associated with a marked decrease in proliferation of NHKs, suggesting that II-4 could selectively expand because of its growth advantage relative to NHKs. Clonal expansion of tumor cells was temporally linked to the decreased expression of filaggrin and keratin 1 expression in adjacent NHKs. These findings indicate that TPA may enable expansion of potentially malignant cells through the epigenetic modification of proliferation in NHKs and differentiation of NHK and II-4 cells.     

A chromodomain protein encoded by the arabidopsis CAO gene is a plant-specific component of the chloroplast signal recognition particle pathway that is involved in LHCP targeting

Adjustment outcomes of 224 transracial and inracial adoptees were investigated using data collected over 17 years. Findings reveal an association between adoptees' outcomes and their race, gender, and adoptive family structure. Placement history was not significant. Implications for policy and practice are discussed, as are future directions for research.     

Use of intrinsic modes in biology: examples of indicial response of pulmonary blood pressure to +/- step hypoxia

Recently, a new method to analyze biological nonstationary stochastic variables has been presented. The method is especially suitable to analyze the variation of one biological variable with respect to changes of another variable. Here, it is illustrated by the change of the pulmonary blood pressure in response to a step change of oxygen concentration in the gas that an animal breathes. The pressure signal is resolved into the sum of a set of oscillatory intrinsic mode functions, which have zero "local mean," and a final nonoscillatory mode. With this device, we obtain a set of "mean trends," each of which represents a "mean" in a definitive sense, and together they represent the mean trend systematically with different degrees of oscillatory content. Correspondingly, the oscillatory content of the signal about any mean trend can be represented by a set of partial sums of intrinsic mode functions. When the concept of "indicial response function" is used to describe the change of one variable in response to a step change of another variable, we now have a set of indicial response functions of the mean trends and another set of indicial response functions to describe the energy or intensity of oscillations about each mean trend. Each of these can be represented by an analytic function whose coefficients can be determined by a least-squares curve-fitting procedure. In this way, experimental results are stated sharply by analytic functions.     

Conformational changes due to vicinal glycosylation: the branched alpha-L-Rhap(1-2)[beta-D-Galp(1-3)]-beta-D-Glc1-OMe trisaccharide compared with its parent disaccharides

Conformations of the alpha-L-Rhap(1-2)-beta-D-Glc1-OMe and beta-D-Galp(1-3)-beta-D-Glc1-OMe disaccharides and the branched title trisaccharide were examined in DMSO-d6 solution by 1H-nmr. The distance mapping procedure was based on rotating frame nuclear Overhauser effect (NOE) constraints involving C- and O-linked protons, and hydrogen-bond constraints manifested by the splitting of the OH nmr signals for partially deuteriated samples. An "isotopomer-selected NOE" method for the unequivocal identification of mutually hydrogen-bonded hydroxyl groups was suggested. The length of hydrogen bonds thus detected is considered the only one motionally nonaveraged nmr-derived constraint. Molecular mechanics and molecular dynamics methods were used to model the conformational properties of the studied oligosaccharides. Complex conformational search, relying on a regular phi, psi-grid based scanning of the conformational space of the selected glycosidic linkage, combined with simultaneous modeling of different allowed orientations of the pendant groups and the third, neighboring sugar residue, has been carried out. Energy minimizations were performed for each member of the phi, psi grid generated set of conformations. Conformational clustering has been done to group the minimized conformations into families with similar values of glycosidic torsion angles. Several stable syn and anti conformations were found for the 1-->2 and 1-->3 bonds in the studied disaccharides. Vicinal glycosylation affected strongly the occupancy of conformational states in both branches of the title trisaccharide. The preferred conformational family of the trisaccharide (with average phi, psi values of 38 degrees, 17 degrees for the 1-->2 and 48 degrees, 1 degree for the 1-->3 bond, respectively) was shown by nmr to be stabilized by intramolecular hydrogen bonding between the nonbonded Rha and Gal residues.     

Determining the imaging quality of intraocular lenses

This paper describes research work motivated by an innovative medical application: computer-assisted transbronchial biopsy. This project involves the registration, with no external localization device, of a preoperative three-dimensional (3-D) computed tomography (CT) scan of the thoracic cavity (showing a tumor that requires a needle biopsy), and an intraoperative endoscopic two-dimensional (2-D) image sequence, in order to provide assistance in transbronchial puncture of the tumor. Because of the specific difficulties resulting from the data being processed, a multilevel strategy was introduced. For each analysis level, the relevant information to process and the corresponding algorithms were defined. This multilevel strategy, thus, provides the best possible accuracy. Original image processing methods were elaborated, dealing with segmentation, registration and 3-D reconstruction of the bronchoscopic images. In particular, these methods involve adapted mathematical morphology tools, a "daemon-based" registration algorithm, and a model-based shape-from-shading algorithm. This pilot study presents the application of these algorithms to recorded bronchoscopic video sequences for five patients. The preliminary results presented here demonstrate that it is possible to precisely localize the endoscopic camera within the CT data coordinate system. The computer can thus synthesize in near real-time the CT-derived virtual view that corresponds to the actual endoscopic view.