Expression profile analysis of colon cancer cells in response to sulindac or aspirin

Nonsteroidal anti-inflammatory drugs (NSAIDs) have a preventive effect against colorectal cancer. Although inhibition of cyclooxygenase-2 plays a crucial role in the suppression of tumors, precise mechanisms of their action remain to be disclosed. To identify genes involved in the growth-suppressive effect of NSAIDs, we utilized cDNA microarray containing 23,040 genes and analyzed time-dependent alteration of gene expression in response to sulindac or aspirin in NSAIDs-sensitive SW480 and SW948 colon-cancer cells as well as in relatively resistant SNU-C4 cells. Consequently we identified 112 genes with commonly altered expression by sulindac and 176 with commonly altered expression by aspirin in the three lines. Addition of sulindac and that of aspirin altered expression levels of 130 and 140 genes, respectively, in SW480 and SW948 cells but not in SNU-C4 cells. These data may lead to a better understanding of growth-suppressive effects on colonic epithelium, and may provide clues for identifying novel therapeutic and/or preventive molecular targets of colon cancer.     

Oxidation-state-dependent protein docking between cytochrome c and cytochrome b(5): high-pressure laser flash photolysis study

To characterize the protein-protein interaction during electron transfer, we used Zn-substituted cytochrome c (ZnCytc) as a model of ferrous Cytc and determined the volume change, DeltaV(d)(Zn), for the dissociation of its complex with ferric cytochrome b(5) (Cytb(5)) by the pressure dependence of its photoinduced electron-transfer kinetics. Under ambient pressure, the dissociation constant, K(d)(Zn), of the ZnCytc/Cytb(5) complex was dependent on the buffer concentration, 1.5 and 12 microM in 2 and 10 mM Tris-HCl, pH 7.4, respectively, which was consistent with formation of salt bridges in its complexation. The dissociation of one salt bridge is usually associated with large volume changes of -10 to -30 cm(3) mol(-1), while pressure dependence of K(d)(Zn) resulted in smaller value of DeltaV(d)(Zn), -8.5 cm(3) mol(-1). Therefore, the interaction between ZnCytc and Cytb(5) cannot be explained only by salt bridge interaction, and the partial cancellation by the positive volume change due to the additional hydrophobic interaction is a plausible explanation for the observed DeltaV(d)(Zn). In addition, DeltaV(d)(Zn) of -8.5 cm(3) mol(-1) was considerably smaller than the previously reported volume change, DeltaV(d)(Fe), of -122 cm(3) mol(-1) in the ferric Cytc/Cytb(5) complex dissociation [Rodgers and Sligar (1991) J. Mol. Biol. 221, 1453-1460]. ZnCytc used here has been assumed to be a reliable model of ferrous Cytc, and thus the discrepancy between our present DeltaV(d)(Zn) and the previous DeltaV(d)(Fe) is discussed on the basis of the protein docking dependent on the oxidation states of heme iron in Cytc.     

Preparation and diastereomeric separation of an (S)- and (R)-1-(methoxycarbonyl)tridec-10-yl glucoside derivative,a precursor for a monosaccharide constituent of resin glycosides

The 6-O-mesyl derivative of phenyl 1-thio-beta-D-glucopyranoside was prepared from D-glucose as a synthetic equivalent of a 6-deoxy-hexosyl donor. Racemic methyl 11-hydroxytetradecanoate (methyl convolvulinolate) was synthesized by Grignard reaction of propylmagnesium bromide with 10-undecenal followed by hydroboration. Both intermediates were coupled by NIS-TfOH-promoted glycosidation to give a mixture of two diasteromeric glucopyranosides, which were separated on a preparative scale by medium pressure chromatography. One of the products was identified as having the natural (S)-configuration by comparison of its 1H NMR spectrum with an authentic sample prepared from the corresponding chiral hydroxyfatty acid.     

Presenilins, the Endoplasmic Reticulum, and Neuronal Apoptosis in Alzheimer's Disease

Abstract: Many cases of autosomal dominant inherited forms of early-onset Alzheimer's disease are caused by mutations in the genes encoding presenilin-1 (PS-1; chromosome 14) and presenilin-2 (PS-2; chromosome 1). PSs are expressed in neurons throughout the brain wherein they appear to be localized primarily to the endoplasmic reticulum (ER) of cell bodies and dendrities. PS-1 and PS-2 show high homology and are predicted to have eight transmembrane domains with the C terminus, N terminus, and a loop domain all on the cytosolic side of the membrane; an enzymatic cleavage of PSs occurs at a site near the loop domain. The normal function of PSs is unknown, but data suggest roles in membrane trafficking, amyloid precursor protein processing, and regulation of ER calcium homeostasis. Homology of PSs to the C. elegans gene sel-12 , which is involved in Notch signaling, and phenotypic similarities of PS-1 and Notch knockout mice suggest a developmental role for PSs in the nervous system. When expressed in cultured cells and transgenic mice, mutant PSs promote increased production of a long form of amyloid β-peptide (Aβ1-42) that may possess enhanced amyloidogenic and neurotoxic properties. PS mutations sensitize cultured neural cells to apoptosis induced by trophic factor withdrawal, metabolic insults, and amyloid β-peptide. The mechanism responsible for the proapoptotic action of mutant PSs may involve perturbed calcium release from ER stores and increased levels of oxidative stress. Recent studies of apoptosis in many different cell types suggest that ER calcium signaling can modulate apoptosis. The evolving picture of PS roles in neuronal plasticity and Alzheimer's disease is bringing to the forefront the ER, an organelle increasingly recognized as a key regulator of neuronal plasticity and survival.     

Heterogeneity of Light Availability and its Effects on Simulated Carbon Gain of Tree Leaves in a Small Gap and the Understory in a Tropical Rain Forest1

To clarify the small-scale heterogeneity of light regimes in a rain forest, photosynthetic photon flux density (PFD) was measured at 1-min intervals during six days at 12 microsites in each of two plots, a small gap and an understory in Pasoh Forest Reserve, Peninsular Malaysia. Frequency distribution of microsite PFD was unimodal with the peak value between 16 and 32 μmol/m²/sec in the small gap, but between 8 and 16 μmol/m²/sec in the understory. In the small gap, PFD was more variable among microsites; total daily PFD and daily sunfleck PFD exceeding 10 μmol/ m²/sec tended to be higher (P <0.05; t-test) compared to those in the understory. Sunfleck PFD exceeding 50 μmol/ m²/sec, however, showed no difference between the two plots. Diffuse PFD transmittance, defined as the ratio of PFD in the forest to that measured at 43 m above ground during the periods 0800-0810 and 1750-1800 h, was significantly higher in the small gap than in the understory plot. Diffuse PFD transmittance was also positively correlated with microsite total daily PFD. To examine the effects of the subtle heterogeneity of light regimes on leaf carbon gain, we simulated carbon gain by sun and shade leaves in a typical shade-tolerant species, Brosimum aticastrum Sw. (Moraceae). Despite the similarity in total daily PFD, total daily carbon gain was considerably higher in the gap than in the understory for both sun and shade leaves. This study suggests that frequency distribution of PFD is critical in describing microsite PFD regimes and determining leaf carbon gain in the tropical forest floor.     

EMG-angle relationship of the hamstring muscles during maximum knee flexion.

The aim of the present study was to investigate the EMG-joint angle relationship during voluntary contraction with maximum effort and the differences in activity among three hamstring muscles during knee flexion. Ten healthy subjects performed maximum voluntary isometric and isokinetic knee flexion. The isometric tests were performed for 5 s at knee angles of 60 and 90 degrees. The isokinetic test, which consisted of knee flexion from 0 to 120 degrees in the prone position, was performed at an angular velocity of 30 degrees /s (0.523 rad/s). The knee flexion torque was measured using a KIN-COM isokinetic dynamometer. The individual EMG activity of the hamstrings, i.e. the semitendinosus, semimembranosus, long head of the biceps femoris and short head of the biceps femoris muscles, was detected using a bipolar fine wire electrode. With isometric testing, the knee flexion torque at 60 degrees knee flexion was greater than that at 90 degrees. The mean peak isokinetic torque occurred from 15 to 30 degrees knee flexion angle and then the torque decreased as the knee angle increased (p<0.01). The EMG activity of the hamstring muscles varied with the change in knee flexion angle except for the short head of the biceps femoris muscle under isometric condition. With isometric contraction, the integrated EMGs of the semitendinosus and semimembranosus muscles at a knee flexion angle of 60 degrees were significantly lower than that at 90 degrees. During maximum isokinetic contraction, the integrated EMGs of the semitendinosus, semimembranosus and short head of the biceps femoris muscles increased significantly as the knee angle increased from 0 to 105 degrees of knee flexion (p<0.05). On the other hand, the integrated EMG of the long head of the biceps femoris muscle at a knee angle of 60 degrees was significantly greater than that at 90 degrees knee flexion with isometric testing (p<0.01). During maximum isokinetic contraction, the integrated EMG was the greatest at a knee angle between 15 and 30 degrees, and then significantly decreased as the knee angle increased from 30 to 120 degrees (p<0.01). These results demonstrate that the EMG activity of hamstring muscles during maximum isometric and isokinetic knee flexion varies with change in muscle length or joint angle, and that the activity of the long head of the biceps femoris muscle differs considerably from the other three heads of hamstrings.     

Immunological characteristics of Hatano high-and low-avoidance rats.

Hatano high- and low-avoidance (HAA and LAA) rats have been genetically selected on the basis of their two-way active avoidance behavior, and have been shown to differ in other behavioral and hormonal parameters. Since close interconnections among the nervous, endocrine and immune systems have been well documented, these two strains might possess differences in aspects of immunological action. In Experiment 1, plasma levels of IgG, IgM, complement 3 (C3), classical pathway hemolytic complement (CH50) and beta(2)-microglobulin were compared between males of the two strains at 5 and 24 weeks of age. Plasma levels of IgG and CH50 were lower in LAA than HAA rats at 5 weeks of age, whereas those differences disappeared at 24 weeks of age. There were no differences between the two strains in plasma levels of IgM, C3 and beta(2)-microglobulin. In Experiment 2, antibody production to sheep red blood cells (SRBC) and mitogen-induced lymphocyte proliferation were compared between 12-week-old males of the two strains. Antibody responses in the PFC assay, plasma anti-SRBC-IgM levels and spleen weights were higher in LAA than HAA rats. LPS-induced lymphocyte proliferation was greater in LAA than HAA rats. It was concluded that HAA rats show earlier development of immunological development, but that antibody production and mitotic response of B lymphocytes may be more pronounced in adult LAA than HAA rats. The strain differences observed in the immunological response may indicate the usefulness of using Hatano rats in studies of behavioral-immunological relationships.     

Identification and Cloning of an mRNA Coding for a Germ Cell-Specific A-Type Lamin in Mice

The nuclear lamins are karyoskeletal proteins which have important functions, such as maintaining nuclear envelope integrity and organizing high order nuclear structure during mitosis in higher eukaryotes. In somatic mammalian cells, the A-type and B-type lamins, composed of lamins A and C and lamins B1 and B2, are major components of the nuclear lamina. However, A-type lamins have as yet not been identified in germ cells and undifferentiated embryonic cells. Here we report the cloning of a new 52-kDa A-type lamin from mouse pachytene spermatocytes, termed lamin C2 because of its similarities with lamin C. It has a sequence identical to that of lamin C except that the N -terminal segment, containing the head and the α-helical coil 1A domains, is replaced with a short non-α-helical stretch of amino acids. In mice, lamin C2 was found to be specifically expressed in germ cells. This specific expression and unique structure suggests a role for lamin C2 in determining the organization of nuclear and chromosomal structures during spermatogenesis.     

Starch-dependent sodium accumulation in the leaves of Vigna riukiuensis

Journal of Plant Research - This research provides insight into a unique salt tolerance mechanism of Vigna riukiuensis. V. riukiuensis is one of the salt-tolerant species identified from the genus...