Matrigel® invasion by the prostate cancer cell lines,PC3 and DU145, and cathepsin L+B activity

Cathepsins L and B are lysosomal cysteine proteinases whose activities and cellular location are altered in many types of cancers and cancer cell lines. Cathepsins L and B play an unspecified role in cancer invasion and metastasis. The purpose of our study was to determine whether cathepsins L and B are important for the ability of two prostate cancer cell lines, PC3 and DU 145, to invade the basement membrane-like preparation, Matrigel®. Exposure of PC3 and DU145 to the irreversible cysteine proteinase inhibitor, E64, decreases the invasive ability of DU145, but not PC3. PC3 and DU145 were treated with the phorbol ester analogue, phorbol 12-myristate 13-acetate (PMA), a known tumor promoter that activates protein kinase C and contributes to the metastatic phenotype. PMA increased secreted cathepsin L+B activity and the invasive ability of PC3 and DU145; co-exposure to E64 and PMA decreased both cathepsin L+B activity and invasion. We conclude that DU145 requires cathepsin L+B activity more than PC3 for the invasion of the Matrigel®. When the amount of secreted cathepsin L+B activity is increased by PMA treatment, however, PC3 becomes dependent on cathepsin L+B for invasion. Our study demonstrates that modulation of the amount of secreted cathepsin L+B activity influences the invasive phenotype of PC3 and DU145.     

Frictional properties of contacting surfaces in the hemelytra-hindwing locking mechanism in the bug<Emphasis Type="Italic"> Coreus marginatus</Emphasis> (Heteroptera,Coreidae)

The structure and function of the hemelytra-to-hindwing locking mechanism of the bug Coreus marginatus were analysed. The system consists of a cuticular protrusion in the ventral side of the hemelytra, which locks the subcostal border of the hindwing in flight. The speed and distance slid by both surfaces against one another during flight were assessed using a combination of high-speed video recordings and a 2D geometrical model. The friction coefficient between sliding surfaces was assessed using a micromanipulator, coupled with force transducers. This was done under three experimental conditions: freshly dissected, air dried and rehydrated ethanol preserved samples. The results showed a high speed of sliding, approximately 0.18 m s^–1, with a relatively low friction coefficient (0.2 ). There was no evident difference in the friction measured under the various treatments, with the exception of the rehydrated condition, which was lower. The surface morphology of the wing locking mechanism, namely outgrowths of one part having rounded edges, and completely flat surface on the counterpart, effectively aids in the reduction of friction at the microscopic level. The structure is effective even dry, and after being preserved in ethanol, suggesting that no cuticle secreted lubrication substance is responsible for its effectiveness. The ultrastructure presumably confers mechanical stability to the system under the high load it is subjected to in flight.     

Water Striders:The Biomechanics of Water Locomotion and Functional Morphology of the Hydrophobic Surface(Insecta:Hemiptera-Heteroptera)

Water striders are insects living on the water surface, over which they can move very quickly and rarely get wetted. We measured the force of free walking in water striders, using a hair attached to their backs and a 3D strain gauge. The error was calculated by comparing force and data derived from geometry and was estimated as 13%. Females on average were stronger (1.32 mN) than males (0.87 mN), however, the ratio of force to weight was not significantly different. Compared with other lighter species, Aquarius paludum seems stronger, but the ratio of force to weight is actually lower. A. paludum applies about 0.3 mN·cm-1 to 0.4 mN·cm-1 with its mid-legs, thus avoiding penetrating the surface tension layer while propelling itself rapidly over the water surface.We also investigated the external morphology with SEM. The body is covered by effectively two layers of macro-and micro-hairs, which renders them hydrophobic. The setae are long (40 um-60 um) and stiff, being responsible for waterproofing, and the microtrichia are much smaller (<10 um), slender, and flexible, holding a bubble over the body when submerged.     

RETRACTED ARTICLE: An accurate method to directly measure water strider’s stroke force on the water (Aquarius paludum: Heteroptera: Gerridae)

We measured the force of free pulling water striders, using a hair attached to their backs and a 3D strain gauge force sensor. We showed the repeatability and accuracy of this method. The error of the method was estimated by comparing the projected angles of the force vector on each plane derived from the force data, with those angles derived from video recordings, and was estimated as 12.4%. Females on average were stronger (1.32 mN) than males (0.87 mN), however the ratio of force/weight was not significantly different. Compared with other lighter species, A. paludum seems to be stronger, but the force/weight ratio is actually lower as expected. A. paludum applies about 0.30–0.40 mN/cm with its mid-legs, thus avoiding penetrating the surface tension while propelling itself rapidly over the water. The corresponding author, Dr Pablo Perez Goodwyn, submitted this article [1] to Central European Journal of Biology (CEJB) shortly after submitting the article [2] to Journal of Bionic Engineering (JBE). JBE published it as a research article in June 2008, and in July 2008 the article was published as a communication in CEJB. Since there are significant unacknowledged similarities between the two papers, it has been brought to the attention of the authors that duplicate submission and publication have taken place. The editors of CEJB consider this an infringement of professional ethics and therefore the decision has been made to retract the article published in Central European Journal of Biology. [1] An accurate method to directly measure water strider’s stroke force on the water (Aquarius paludum: Heteroptera: Gerridae), Central European Journal of Biology, Vol. 3, No. 3, 2008, pp. 299–306 [2] Water striders: the biomechanics of water locomotion and functional morphology of the hydrophobic surface (Insecta: Hemiptera-Heteroptera), Journal of Bionic Engineering, Vol. 5, No. 2, 2008, pp. 121-126     

Attachment ability of the southern green stink bug, <Emphasis Type="Italic">Nezara viridula</Emphasis> (L.), on plant surfaces

Traction forces of male and female southern green stink bugs Nezara viridula (L.) were measured on adaxial leaves of green beans (Phaseolus vulgaris L. cv. ‘Nagauzura’), peas (Pisum sativum L. cv. ‘Hakuryu’), rice (Oryza sativa L. cv. ‘Mac hin sung’), and soybean (Glycine max [L.] Merr. cv. ‘Enrei’), as well as on glass as a control surface. Bugs attached well on three-dimensionally structured surfaces covered with anti-adhesive epicuticular wax crystals. Strongest pulls were generated on adaxial green bean leaves, corresponding to safety factors (traction force/body weight) of 11.1 and 11.6 in males and females, respectively. These values were slightly lower on soybean (males: 7.4, females: 8.0) and rice leaves (males: 8.9, females: 10.6). Trichomes and papillae are assumed to promote pentatomid bug’s attachment. On tabular, wax-covered pea leaves, safety factors decreased significantly to 1.7 and 1.6, in males and females, respectively. Differently, on non-structured glass, safety factors resembled those on rice and soybean leaves (males: 9.6, females: 8.0). No statistical differences in traction force and safety factor between sexes were detected on any substrate. Surface wettability did not significantly affect the results. Both robust claws and tough adhesive pads enable the N. viridula bugs to grasp and adhere to a wide range of various plant substrates, including such ones covered with anti-adhesive wax crystals. However, tabular, pruinose pea leaves were detected to prevent the foothold of the southern green stink bugs, although pea is known as one of their common host plants.     

Matrigel invasion by the prostate cancer cell lines, PC3 and DU145, and cathepsin L+B activity.

Cathepsins L and B are lysosomal cysteine proteinases whose activities and cellular location are altered in many types of cancers and cancer cell lines. Cathepsins L and B play an unspecified role in cancer invasion and metastasis. The purpose of our study was to determine whether cathepsins L and B are important for the ability of two prostate cancer cell lines, PC3 and DU 145, to invade the basement membrane-like preparation, Matrigel. Exposure of PC3 and DU145 to the irreversible cysteine proteinase inhibitor, E64, decreases the invasive ability of DU145, but not PC3. PC3 and DU145 were treated with the phorbol ester analogue, phorbol 12-myristate 13-acetate (PMA), a known tumor promoter that activates protein kinase C and contributes to the metastatic phenotype. PMA increased secreted cathepsin L+B activity and the invasive ability of PC3 and DU145; co-exposure to E64 and PMA decreased both cathepsin L+B activity and invasion. We conclude that DU145 requires cathepsin L+B activity more than PC3 for the invasion of the Matrigel. When the amount of secreted cathepsin L+B activity is increased by PMA treatment, however, PC3 becomes dependent on cathepsin L+B for invasion. Our study demonstrates that modulation of the amount of secreted cathepsin L+B activity influences the invasive phenotype of PC3 and DU145.     

Gripping ease in southern green stink bugs Nezara viridula L. (Heteroptera: Pentatomidae): Coping with geometry,orientation and surface wettability of substrate

The southern green stink bug Nezara viridula L. (Heteroptera, Pentatomidae) is highly polyphagous, preferring apically situated seeds and fruits on more than 150 plant species belonging to over 30 plant families all over the world. This forces them to move over highly variable terrains, including plant stems, leaves, pods and buds, which requires efficient attachment. Stink bugs have long slender legs and feet (tarsi) equipped with paired curved claws, paired soft adhesive pads (pulvilli), and flattened lanceolate hairs (setae), which arise ventrally on the first and second foot segments (tarsomeres). To characterize their attachment abilities on well‐defined test substrates, here we comparatively measured and analyzed the traction forces of bugs walking horizontally and vertically on hydrophilic (water attractive) and hydrophobic (water repellent) glass plates and rods. The latter correspond to the geometry of preferred feeding sites of stink bugs in the field. The results show a clear contribution of tarsal flattened lanceolate hairs to the stink bug's attachment. Higher traction forces are generated on a glass rod than on a glass plate, corresponding to up to individual maximum of 43 times the stink bug's body weight. Substrate hydrophobicity promotes the attachment, while the measured forces are up to eight times lower when tarsal hairs are disabled. The combination of smooth and hairy tarsal pads results in a remarkable attachment ability, which enables N. viridula to climb unstable apical plant parts, and supports their invasive behavior and global dispersion.     

Semiaquatic Heteroptera locomotion: coral treaders (Hermatobates weddi, Hermatobatidae), sea skaters (Halovelia septentrionalis, Veliidae), and water striders (Metrocoris histrio, Gerridae). Usual and unusual gaits

Using high-speed video recordings, we carried out an analysis of the locomotion gaits of the following aquatic Heteroptera: coral treaders Hermatobates weddi (Hermatobatidae), sea striders Halovelia septentrionalis (Veliidae), and water striders Metrocoris histrio (Gerridae), in the Island of Amami Oshima, Kagoshima Prefecture, Japan. Most insects use an alternating double tripod gait for walking, whereas species of Gerridae and some Veliidae use a synchronous rowing gait. We found that H. weddi used a peculiar locomotion gait, a modification of the double tripod gait. In this special gait, two alternating dipods (mid and hind legs) are used, while the forelegs remained inactive. Contralateral mid and hind stroked simultaneously. The mid leg recovered immediately after the stroke; however, the hind leg was delayed and remained extended after the stroke. Next, the following bipod stroked, and when that mid leg finished the stroke, both ipsilateral mid and hind (the one which did not recover after the stroke) legs recovered together. Turning is also unique in H. weddi because the body axis rotation and the course turning (deflection) were clearly separated in two phases. We compared the kinematics of H. weddi pattern with the synchronous rowing pattern found in H. septentrionalis and M. histrio and discussed some biomechanical consequences. We also analyzed phylogenetic implications of this gait, and we posit that the modified double dipod gait is a uniquely derived character of the family Hermatobatidae. The synchronous rowing gait would be an autapomorphy for the clade Gerridae + Veliidae. The modified thorax, with the meso and metacoxae horizontally directed, would be a synapomorphy for the superfamily Gerroidea (Hermatobatidae, Gerridae, and Veliidae). Handling editor: Koen Martens     

Reconstitution of membrane proteins: sequential incorporation of integral membrane proteins into preformed lipid bilayers

Several integral membrane proteins can be inserted sequentially into preformed unilamellar vesicles (ULV's) composed of dimyristoylphosphatidylcholine (DMPC) and cholesterol in a gel phase. Thus, proteoliposomes of DMPC, cholesterol, and bacteriorhodopsin from Halobacterium halobium rapidly incorporate UDPglucuronosyltransferase (EC 2.4.1.17) from pig liver microsomes, cytochrome oxidase from beef heart mitochondria, and additional bacteriorhodopsin, added sequentially. This process of spontaneous incorporation can be regulated to produce complex artificial membranes that contain phospholipids and proteins at ratios (mol/mol) equivalent to what is found in biological membranes. The ability of the lipid-protein bilayers to incorporate additional integral membrane proteins is not affected by annealing of the proteoliposomes at 37 degrees C nor by the order of addition of the proteins. Bacteriorhodopsin-containing vesicles formed by the sequential addition of integral membrane proteins demonstrate light-driven proton pumping. Therefore, they have retained a vesicular structure. Vesicles containing one or two different proteins will fuse with each other at 21 degrees C or with ULV's devoid of proteins. Incorporation of bacteriorhodopsin or UDPglucuronosyltransferase into proteoliposomes containing DMPC, with or without cholesterol as impurity, also occurs above the phase transition for DMPC. The presence of a protein in a liquid-crystalline bilayer provides the necessary condition for promoting the spontaneous incorporation of other membrane proteins into preformed bilayers.