Extra-nuclear signaling of progesterone receptor to breast cancer cell movement and invasion through the actin cytoskeleton

Progesterone plays a role in breast cancer development and progression but the effects on breast cancer cell movement or invasion have not been fully explored. In this study, we investigate the actions of natural progesterone and of the synthetic progestin medroxyprogesterone acetate (MPA) on actin cytoskeleton remodeling and on breast cancer cell movement and invasion. In particular, we characterize the nongenomic signaling cascades implicated in these actions. T47-D breast cancer cells display enhanced horizontal migration and invasion of three-dimensional matrices in the presence of both progestins. Exposure to the hormones triggers a rapid remodeling of the actin cytoskeleton and the formation of membrane ruffles required for cell movement, which are dependent on the rapid phosphorylation of the actin-regulatory protein moesin. The extra-cellular small GTPase RhoA/Rho-associated kinase (ROCK-2) cascade plays central role in progesterone- and MPA-induced moesin activation, cell migration and invasion. In the presence of progesterone, progesterone receptor A (PRA) interacts with the G protein G alpha(13), while MPA drives PR to interact with tyrosine kinase c-Src and to activate phosphatidylinositol-3 kinase, leading to the activation of RhoA/ROCK-2. In conclusion, our findings manifest that progesterone and MPA promote breast cancer cell movement via rapid actin cytoskeleton remodeling, which are mediated by moesin activation. These events are triggered by RhoA/ROCK-2 cascade through partially differing pathways by the two compounds. These results provide original mechanistic explanations for the effects of progestins on breast cancer progression and highlight potential targets to treat endocrine-sensitive breast cancers.     

Adrenomedullin is increased by pulsatile shear stress on the vascular endothelium via periodic acceleration (pGz)

Periodic acceleration (pGz) is produced by a platform which moves the supine body repetitively in a headward to footward direction. The imparted motion produces pulsatile shear stress on the vascular endothelium. Pulsatile shear stress on the vascular endothelium has been shown to elicit production of a host of cardioprotective, cytoprotective mediators. The purpose of this study was to ascertain if pGz also enhances production of adrenomedullin (AM) in normal healthy swine. Twelve pigs (weight range 20-30 kg) were anesthetized, intubated and placed on conventional mechanical ventilation. All animals were secured to the motion platform. In one group (pGz) (n=7) was activated for 1h, and monitored for an additional 3h. A control group (CONT) (n=5) served as time control. Arterial blood gases, hemodynamic measurements, and serum for AM, interleukin 4, 6 and thromboxane B(2) (TBXB2) were measured at baseline, immediately after pGz, and 3h after pGz had been discontinued. There was no significant change from baseline value in IL-4, IL-6 or TBXB2. Mean arterial blood pressure decreased in pGz-treated animals from 115+/-10 at baseline to 90+/-8 after 60 min of pGz (p<0.01). AM levels increase from 776+/-176 pg/ml baseline to 1160+/-68 pg/ml immediately after pGz, and remained elevated to 1584+/-160 pg/ml, 3h after pGz (p<0.01 vs. BL). This is the first report of AM-enhanced production using a non-invasive method of increasing pulsatile shear stress on the vascular endothelium. pGz increases production of AM in normal healthy swine. These changes are independent of IL-4, IL-6 or TBXB2 production.     

Biological aspects of <Emphasis Type="Italic">Amblyomma brasiliense</Emphasis> (Acari: Ixodidae) under laboratory conditions

Although Amblyomma brasiliense Aragão 1908 has been reported as one of the most aggressive ticks to humans in Brazil, information about the biology of this tick species is virtually inexistent. This work reports data on the life cycle of A. brasiliense fed on rabbits and pigs and maintained in an incubator at 20°C, 90% RH and 12 h of light for off-host development. Tick yield of adult females fed on pigs and rabbits was 81.2% and 58.3%, respectively. Females fed on pigs had mean engorgement weight of 862.3 mg and egg mass of 208 mg, while females fed on rabbits had mean engorgement weight of 606.1 mg and egg mass of 160 mg; these values did not differ statistically between host species. Feeding period of female ticks fed on pigs (10 days) was significantly shorter than that on rabbits (17 days). Mean preoviposition period was slightly longer (35.9 days) for ticks fed on pigs than on rabbits (30 days). The minimum incubation period of eggs of ticks from both host species was similar and over 100 days. Egg production efficiency was low for females fed on both hosts (less than 30% and 20% for ticks from pigs and rabbits, respectively). More than 55% of larvae and 79% of nymphs fed on rabbits, set free inside the feeding chambers, engorged successfully. These ticks attained an engorgement weight of 1.3 and 18.2 mg, respectively, and fed for approximately 5 days. The minimum pre-molt period was 30 days for engorged larvae and over 44 days for nymphs. Molting success was low, less than 50% in the case of larvae and less than 20% for nymphs. Further studies are required to better determine the off-host requirements of this tick species.     

Structural characterization of the cell wall D-glucans isolated from the mycelium of Botryosphaeria rhodina MAMB-05

Three D-glucans were isolated from the mycelium of the fungus Botryosphaeria rhodina MAMB-05 by sequential extraction with hot-water and hot aqueous KOH (2% w/v) followed by ethanol precipitation. Following their purification by gel permeation chromatography on Sepharose CL-4B, the structural characteristics of the D-glucans were determined by FT-IR and 13C NMR spectroscopy and, after methylation, by GC-MS. The hot-water extract produced a fraction designated Q1A that was a beta-(1-->6)-D-glucan with the following structure: [Formula: see text] The alkaline extract, when subjected to repeated freeze-thawing, yielded two fractions: K1P (insoluble) that comprised a beta-(1-->3)-D-glucan with beta-D-glucose branches at C-6 with the structure: [Formula: see text] and K1SA (soluble) consisting of a backbone chain of alpha-(1-->4)-linked D-glucopyranosyl residues substituted at O-6 with alpha-D-glucopyranosyl residues: [Formula: see text]     

[NiFe] hydrogenase from Desulfovibrio desulfuricans ATCC 27774: gene sequencing, three-dimensional structure determination and refinement at 1.8 Å and modelling studies of its interaction with the tetrahaem cytochrome c 3

The primary and three-dimensional structures of a [NiFe] hydrogenase isolated from D. desulfitricans ATCC 27774 were determined, by nucleotide analysis and single-crystal X-ray crystallography. The three-dimensional structural model was refined to R=0.167 and Rfree=0.223 using data to 1.8 A resolution. Two unique structural features are observed: the [4Fe-4S] cluster nearest the [NiFe] centre has been modified [4Fe-3S-3O] by loss of one sulfur atom and inclusion of three oxygen atoms; a three-fold disorder was observed for Cys536 which binds to the nickel atom in the [NiFe] centre. Also, the bridging sulfur atom that caps the active site was found to have partial occupancy, thus corresponding to a partly activated enzyme. These structural features may have biological relevance. In particular, the two less-populated rotamers of Cys536 may be involved in the activation process of the enzyme, as well as in the catalytic cycle. Molecular modelling studies were carried out on the interaction between this [NiFe] hydrogenase and its physiological partner, the tetrahaem cytochrome c3 from the same organism. The lowest energy docking solutions were found to correspond to an interaction between the haem IV region in tetrahaem cytochrome c3 with the distal [4Fe-4S] cluster in [NiFe] hydrogenase. This interaction should correspond to efficient electron transfer and be physiologically relevant, given the proximity of the two redox centres and the fact that electron transfer decay coupling calculations show high coupling values and a short electron transfer pathway. On the other hand, other docking solutions have been found that, despite showing low electron transfer efficiency, may give clues on possible proton transfer mechanisms between the two molecules.     

Distribution and migration of Mississippi Kites in South America

Mississippi Kites (Ictinia mississippiensis) are trans‐equatorial, long‐distance migrants that breed in North America and overwinter in South America. Information about their migration routes and winter distribution in the Neotropics is mostly anecdotal. By compiling records of Mississippi Kites in South America from the literature and previously unpublished observations (1904–2010), we identified 96 locality records (a location where a flock or individual was recorded) and 146 independent records of flocks (observations of flocks irrespective of year, location, or time of year). Our locality records included 38 (39%) during southbound migration (1 September–30 November), 18 (19%) during northbound migration (15 February–30 April), 38 (39%) during austral summer (1 December–14 February), and two (3%) during austral winter (1 May–31 August). Most Mississippi Kites (84, 88%) were observed between the 11°S and 32°S latitudinal band in central South America. Of our independent records of flocks, 133 (92%) were observed between 11°S and 32°S, 12 (7%) between 11°N and 10°S, and a lone vagrant (1%) between 33°S and 43°S. Our data suggest that Mississippi Kites are common and widespread in the austral summer between 11°S and 32°S in central South America. On the basis of the number of locality records (N = 52, 54%) and number of flocks of Mississippi Kites observed between 22°S and 32°S (N = 61, 42%), the Chaco forest appears to be the main wintering grounds for the species. However, additional monitoring is needed to further test this hypothesis. A large portion of Chaco habitat is now under cultivation, and how this habitat transformation might influence the annual cycle of Mississippi Kites is unknown.     

Gender Differences in Emotional Responses to Cooperative and Competitive Game Play

Previous research indicates that males prefer competition over cooperation, and it is sometimes suggested that females show the opposite behavioral preference. In the present article, we investigate the emotions behind the preferences: Do males exhibit more positive emotions during competitive than cooperative activities, and do females show the opposite pattern? We conducted two experiments where we assessed the emotional responses of same-gender dyads (in total 130 participants, 50 female) during intrinsically motivating competitive and cooperative digital game play using facial electromyography (EMG), skin conductance, heart rate measures, and self-reported emotional experiences. We found higher positive emotional responses (as indexed by both physiological measures and self-reports) during competitive than cooperative play for males, but no differences for females. In addition, we found no differences in negative emotions, and heart rate, skin conductance, and self-reports yielded contradictory evidence for arousal. These results support the hypothesis that males not only prefer competitive over cooperative play, but they also exhibit more positive emotional responses during them. In contrast, the results suggest that the emotional experiences of females do not differ between cooperation and competition, which implies that less competitiveness does not mean more cooperativeness. Our results pertain to intrinsically motivated game play, but might be relevant also for other kinds of activities.