Reciprocal changes in gene expression profiles of cocultured breast epithelial cells and primary fibroblasts

The importance of epithelial-stroma interaction in normal breast development and tumor progression has been recognized. To identify genes that were regulated by these reciprocal interactions, we cocultured a nonmalignant (MCF10A) and a breast cancer derived (MDA-MB231) basal cell lines, with fibroblasts isolated from breast benign-disease adjacent tissues (NAF) or with carcinoma-associated fibroblasts (CAF), in a transwell system. Gene expression profiles of each coculture pair were compared with the correspondent monocultures, using a customized microarray. Contrariwise to large alterations in epithelial cells genomic profiles, fibroblasts were less affected. In MDA-MB231 highly represented genes downregulated by CAF derived factors coded for proteins important for the specificity of vectorial transport between ER and golgi, possibly affecting cell polarity whereas the response of MCF10A comprised an induction of genes coding for stress responsive proteins, representing a prosurvival effect. While NAF downregulated genes encoding proteins associated to glycolipid and fatty acid biosynthesis in MDA-MB231, potentially affecting membrane biogenesis, in MCF10A, genes critical for growth control and adhesion were altered. NAFs responded to coculture with MDA-MB231 by a decrease in the expression of genes induced by TGF beta 1 and associated to motility. However, there was little change in NAFs gene expression profile influenced by MCF10A. CAFs responded to the presence of both epithelial cells inducing genes implicated in cell proliferation. Our data indicate that interactions between breast fibroblasts and basal epithelial cells resulted in alterations in the genomic profiles of both cell types which may help to clarify some aspects of this heterotypic signaling. (C) 2009 UICC

Clinical correlates of eating disorder comorbidity in women with bipolar disorder type I

Objective: To report on the presence of current and lifetime eating disorders (ED) in a well-defined sample of 137 female individuals with bipolar disorder type I. Methods: Trained psychiatrists interviewed the patients, and the diagnoses of BD and comorbidities were confirmed using the Structured Clinical Interview for the Diagnostic and Statistical Manual of Mental Disorders, Axis I Disorders. Clinical and demographic characteristics of both groups (group with ED vs. group without ED) were compared. Results: Female patients with ED had an earlier onset of BD and an increased number of mood episodes, predominantly depressive. Women in the ED group also had higher rates of comorbidity with substance use disorders and anxiety disorders and reported a history of suicide attempts more frequently than women without ED. Conclusion: The presence of ED is a correlate of severity of BD type 1, and interventions should be developed to minimize distress and suicide risk and to improve treatment outcome. (C) 2010 Elsevier B.V. All rights reserved.

STANDARDIZATION OF SOME ELECTROCARDIOGRAPHIC PARAMETERS OF CAPTIVE LEOPARD CATS (LEOPARDUS TIGRINUS)

Thirty-three captive leopard cats, Leopardus tigrinus, were anesthetized with xylazine (1-2 mg/kg) and ketamine (10 mg/kg), and electrocardiograph (ECG) tests were recorded in all leads with 1 cm = 1 mV sensibility and 25 mm/sec speed repeating DII lead at 50 mm/sec speed with the same sensibility. Results expressed by mean and standard deviation were: heart rate (HR) = 107 +/- 17 (bpm); P-wave = 0.048 +/- 0.072 (s) x 0.128 +/- 0.048 (mV); PR interval = 0.101 +/- 0.081 (s); QRS compound = 0.053 +/- 0.012 (s) x 1.446 +/- 0.602 (mV); QT interval = 0.231 +/- 0.028 (s); R-wave (CV(6)LL) = 1.574 +/- 0.527 (mV); R-wave (CV(6)LU) = 1.583 +/- 0.818 (mV); heart rhythm: normal sinus rhythm (15.2%), sinus rhythm with wandering pacemaker (WPM) (60.6%), sinus arrhythmia with WPM (24.2%); electric axis: between +30 degrees and +60 degrees (6.1%), +60 (6.1%), between +60 degrees and +90 degrees (57.6%), +90 degrees (9%), between +90 degrees and +120 degrees (21.2%); ST segment: normal (75.7%), elevation (18.2%), depression (6.1%); T-wave polarity (DII): positive (100%); T-wave (V(10)): absent (6.1%), negative (63.6%), positive (18.2%), and with interference (12.1%). Through ECG data comparison with other species, unique features of Leopardus tigrinus` (leopard cat) ECG parameters were detected. Some of the study animals presented with an R-Wave amplitude that was indicative of left ventricle overload according to patterns for normal domestic cats (Felis cati). Echocardiographic exams revealed normal heart cavities` function and morphology. The aim of this study was to establish some electrocardiographic parameters of captive L. tigrinus.

Oral squamous cell carcinoma: status of tight junction claudins in the different histopathological patterns and relationship with clinical parameters. A tissue-microarray-based study of 136 cases

Aims Claudins are integral transmembrane proteins of the tight junctions, critical for maintaining cell adhesion and polarity. Alterations in the expression of individual claudins have been detected in carcinomas and appear to correlate with tumour progression. Methods In this study, a panel of anti-claudin antibodies (anti-claudins 1, 2, 3, 4, 5 and 7) was employed to map claudin expression in 136 cases of oral squamous cell carcinoma (OSCC) organised in a tissue microarray. Results Claudins were expressed in a reticular pattern up to the prickle layer in normal mucosal epithelium. In OSCC, claudins were strongly present in well-differentiated tumours, they presented mild and low expression in moderately differentiated OSCC, and were negative in poorly differentiated OSCC; the absences of claudin 1 (p = 0.002) and claudin 4 (p<0.001) were associated with moderately/poorly differentiated tumours. Strong expression of claudin 4 was associated with decreased perineural infiltration (p = 0.024). Claudins 5 and 7 were mostly negative or weakly expressed in all cases studied. Expression of claudin 7 was associated with the early clinical stages of the disease, whereas loss of claudin 7 tended to be more frequent in advanced stages of OSCC (p = 0.054). Absence of claudin 7 was also associated with absent vascular infiltration (p = 0.045) and with presence of recurrence (p = 0.052). Conclusions Claudin expression patterns showed a strong correlation with histological type of OSCC; claudin expression was decreased in areas of invasion, and negative in poorly differentiated tumours. This pattern may be related to evolution and prognosis of these tumours, especially in the case of claudin 7, which seems to be associated with a poor prognosis in OSCC.

Extracts from pitanga (Eugenia uniflora L.) leaves: Influence of extraction process on antioxidant properties and yield of phenolic compounds

Different extraction processes were employed to extract the polyphenolic compounds from pitanga (Eugenia uniflora L) leaves: a one-step process using water, ethanol or supercritical CO(2) as solvents, and a two-step process using supercritical CO(2) followed by either water or ethanol. The total polyphenolic compounds, total flavonoids and antioxidant activity were determined in all the extracts obtained. The process performance was evaluated with respect to three variables: global extraction yield, concentration and yield of both polyphenols and flavonoids in the extracts. For the one-step extraction, the results showed that the extraction yield increased with solvent polarity. For the two-step process, the results suggested that water was more efficient in extracting the phenolic compounds from E. uniflora when the matrix was previously extracted with scCO(2). With respect to the antioxidant activity, the ethanolic extracts obtained from both processes, using either the DPPH radical scavenging method or the beta-carotene bleaching method, presented high antioxidant activities. (C) 2010 Elsevier B.V. All rights reserved.

Paleomagnetism and rock magnetism of the Neoproterozoic Itajai Basin of the Rio de la Plata craton (Brazil): Cambrian to Cretaceous widespread remagnetizations of South America

A detailed rock magnetic and paleomagnetic study was performed on samples from the Neoproterozoic Itajai Basin in the state of Santa Catarina, Brazil, in order to better constrain the paleogeographic evolution of the Rio de la Plata craton between 600 and 550 Ma. However, rock magnetic properties typical of remagnetized rocks and negative response in the fold test indicated that these rocks carried a secondary chemical remanent magnetization. After detailed AF and thermal cleaning, almost all samples showed a normal polarity characteristic remanent magnetization component close to the present geomagnetic field. The main magnetic carriers are magnetite and hematite, probably of authigenic origin. The mean paleomagnetic pole of the ltajai Basin is located at Plat= -84 degrees, Plong = 97.5 degrees (A95 = 2 degrees) and overlaps the lower Cretaceous segment of the apparent polar wander path of South America, suggesting a cause and effect with the opening of the South Atlantic Ocean. A compilation of remagnetized paleomagnetic poles from South America is presented that highlights the superposition of several large-scale remagnetization events between the Cambrian and the Cretaceous. It is suggested that some paleomagnetic poles used to calibrate the APWP of Gondwana at Precambrian times need to be revised; the indication of remagnetized areas in southern South America may offer some help in the selection of sites for future paleomagnetic investigations in Precambrian rocks. (C) 2011 International Association for Gondwana Research. Published by Elsevier B.V. All rights reserved.

Palaeomagnetism, rock magnetism and AMS of the Cabo Magmatic Province, NE Brazil, and the opening of South Atlantic

P>Reconstruction of the South Atlantic opening has long been a matter of debate and several models have been proposed. One problem in tracing properly the Atlantic history arises from the existence of a long interval without geomagnetic reversals, the Cretaceous Normal Superchron, for which ages are difficult to assign. Palaeomagnetism may help in addressing this issue if high-quality palaeomagnetic poles are available for the two drifting continental blocks, and if precise absolute ages are available. In this work we have investigated the Cabo Magmatic Province, northeastern Brazil, recently dated at 102 +/- 1 Ma (zircon fission tracks, Ar39/Ar40). All volcanic and plutonic rocks showed stable thermal and AF demagnetization patterns, and exhibit primary magnetic signatures. AMS data also support a primary origin for the magnetic fabric and is interpreted to be contemporaneous of the rock formation. The obtained pole is located at 335.9 degrees E/87.9 degrees S (N = 24; A(95) = 2.5; K = 138) and satisfies modern quality criteria, resulting in a reference pole for South America at similar to 100 Ma. This new pole also gives an insight to test and discuss the kinematic models currently proposed for the South Atlantic opening during mid-Cretaceous.

Paleointensity data from Early Cretaceous Ponta Grossa dikes (Brazil) using a multisample method

Definition of the long-term variation of the geomagnetic virtual dipole moment requires more reliable paleointensity results. Here, we applied a multisample protocol to the study of the 130.5 Ma Ponta Grossa basaltic dikes (southern Brazil) that carry a very stable dual-polarity magnetic component. The magnetic stability of the samples wits checked using thermomagnetic curves and by monitoring the magnetic Susceptibility evolution through the paleointensity experiments. Twelve sites containing the least alterable samples were chosen for the paleointensity measurements. Although these rocks failed stepwise double-heating experiments, they yielded coherent results in the multisample method for all sites but one. The coherent sites show low to moderate field intensities between 5.7 +/- 0.2 and 26.4 +/- 0.7 mu T (average 13.4 +/- 1.9 mu T). Virtual dipole moments for these sites range from 1.3 +/- 0.04 to 6.0 +/- 0.2 x 10(22) A m(2) (average 2.9 +/- 0.5 x 10(22) A m(2)). Our results agree with the tendency for low dipole moments during the Early Cretaceous, immediately prior to the Cretaceous Normal Superchron (CNS). The available paleointensity database shows a strong variability of the field between 80 and 160 Ma. There seems to be no firm evidence for a Mesozoic Dipole Low, but a long-term tendency does emerge from the data with the highest dipole moments Occurring at the middle of the CNS.

A late Neoproterozoic paleomagnetic pole for the Congo craton: Tectonic setting, paleomagnetism and geochronology of the Nola dike swarm (Central African Republic)

New structural, geochronological and paleomagnetic data were obtained on dolerite dikes of the Nola region (Central African Republic) at the northern border of the Congo craton. In this region metavolcanic, successions were thrust southward onto the craton during the Panafrican orogenic events. Our structural data reveal at least two structural klippes south of the present-day limits of the Panafrican nappe suggesting that it has once covered the whole Nola region, promoting the pervasive hydrothermal green-schist metamorphism observed in the underlying cratonic basement and also in the intrusive dolerite dikes. Paleomagnetic measurements revealed a stable dual-polarity low-inclination magnetization component in nine dikes (47 samples), carried by pyrrhotite and magnetite. This component corresponds to a paleopole at 304.8 degrees E and 61.8 degrees S (dp = 5.4, dm = 10.7) graded at 2 = 6. Both metamorphism and magnetic resetting were dated by the Ar-40/Ar-39 method on amphibole grains separated from the dikes at 571 +/- 6 Ma. The Nola pole is the first well-dated paleomagnetic pole for the Congo craton between 580 and 550 Ma. It marks a sudden change in direction of the Congo craton apparent polar wander path at the waning stages of the Panafrican orogenic events. (c) 2008 Elsevier B.V. All rights reserved.

Programs to compute magnetization to density ratio and the magnetization inclination from 3-D gravity and magnetic anomalies

Vector field formulation based on the Poisson theorem allows an automatic determination of rock physical properties (magnetization to density ratio-MDR-and the magnetization inclination-MI) from combined processing of gravity and magnetic geophysical data. The basic assumptions (i.e., Poisson conditions) are: that gravity and magnetic fields share common sources, and that these sources have a uniform magnetization direction and MDR. In addition, the previously existing formulation was restricted to profile data, and assumed sufficiently elongated (2-D) sources. For sources that violate Poisson conditions or have a 3-D geometry, the apparent values of MDR and MI that are generated in this way have an unclear relationship to the actual properties in the subsurface. We present Fortran programs that estimate MDR and MI values for 3-D sources through processing of gridded gravity and magnetic data. Tests with simple geophysical models indicate that magnetization polarity can be successfully recovered by MDR-MI processing, even in cases where juxtaposed bodies cannot be clearly distinguished on the basis of anomaly data. These results may be useful in crustal studies, especially in mapping magnetization polarity from marine-based gravity and magnetic data. (c) 2007 Elsevier Ltd. All rights reserved.

Scale architecture of the palmar and plantar epidermis of Polychrus acutirostris Spix, 1825 (Iguania, Polychrotidae) and its relationship to arboreal locomotion

In tetrapod squamates, the diversity of micro-ornamentations of the epidermis of the contact areas of hands and feet is generally associated with constraints and modalities related to locomotion. Polychrus acutirostris is a medium-sized lizard that occurs in open heterogeneous habitats in South America, such as the cerrados, caatingas, and fallow lands. It progresses slowly on branches of various diameters in its arboreal environment. It can also move more rapidly on the ground. The hands and feet are prehensile and may be considered an adaptation for grasping and climbing. Epidermal surfaces from the palmar and plantar areas of the hands and feet of P. acutirostris were prepared for SEM examination, and studied at various magnifications. They show three major levels of complexity: (1) scale types, organized in gradients of size and imbrication, (2) scalar ornamentations, organized by increasing complexity and polarity, and (3) presence of Oberhautchen showing typically iguanian honeycomb micro-ornamentations. The shape and surface structure of the scales with their pattern of micro-ornamental peaks, which improve grip, and the grasping hands and feet indicate that P. acutirostris is morpho-functionally specialized for arboreality. (C) 2009 Elsevier GmbH. All rights reserved.

Histological and functional renal alterations caused by Bothrops alternatus snake venom: Expression and activity of Na(+)/K(+)-ATPase

Background: Acute renal failure is a serious complication of human envenoming by Bothrops snakes. The ion pump Na(+)/K(+)-ATPase has an important role in renal tubule function, where it modulates sodium reabsorption and homeostasis of the extracellular compartment. Here, we investigated the morphological and functional renal alterations and changes in Na(+)/K(+)-ATPase expression and activity in rats injected with Bothrops alternatus snake venom. Methods: Male Wistar rats were injected with venom (0.8 mg/kg, iv.) and renal function was assessed 6.24, 48 and 72 h and 7 days post-venom. The rats were then killed and renal Na(+)/K(+)-ATPase activity was assayed based on phosphate release from ATP; gene and protein expressions were assessed by real time PCR and immunofluorescence microscopy, respectively. Results: Venom caused lobulation of the capillary tufts, dilation of Bowman`s capsular space. F-actin disruption in Bowman`s capsule and renal tubule brush border, and deposition of collagen around glomeruli and proximal tubules that persisted seven days after envenoming. Enhanced sodium and potassium excretion, reduced proximal sodium reabsorption, and proteinuria were observed 6 h post-venom, followed by a transient decrease in the glomerular filtration rate. Gene and protein expressions of the Na(+)/K(+)-ATPase alpha(1) subunit were increased 6 h post-venom, whereas Na(+)/K(+)-ATPase activity increased 6 h and 24 h post-venom. Conclusions: Bothrops alternatus venom caused marked morphological and functional renal alterations with enhanced Na(+)/K(+)-ATPase expression and activity in the early phase of renal damage. General significance: Enhanced Na(+)/K(+)-ATPase activity in the early hours after envenoming may attenuate the renal dysfunction associated with venom-induced damage. (C) 2011 Elsevier B.V. All rights reserved.

Distribution of small Rho GTPases in the developing rat submandibular gland

During the rat submandibular gland (SMG) development, organogenesis and cytodifferentiation depend on the actin cytoskeleton, which is regulated by small Rho GTPases. These proteins link cell surface receptors to pathways that regulate cell motility, polarity, gene expression, vesicular trafficking, proliferation and apoptosis. The aim of this study was to evaluate, by immunohistochemistry, the distribution pattern of RhoA, RhoB, RhoC, Rac1 and Cdc42 during cytodifferentiation of the rat SMG and in male adults. All GTPases were found in epithelial and mesenchymal tissues throughout gland development. Rac1 appeared to be important for parenchyma expansion at the beginning of cytodifferentiation, while RhoC, Cdc42 and the inactive phosphorylated form of Rac1 seemed associated with lumen formation and cell polarization in terminal tubules. RhoA and RhoB labeling was evident throughout development. All GTPases were differentially expressed in the adult gland, suggesting that they play specific roles during differentiation and function of the rat SMG.

Structural characterization of the interaction of the polyene antibiotic Amphotericin B with DODAB bicelles and vesicles

Amphotericin B (AmB) is widely used in the treatment of systemic fungal infections, despite its toxic effects. Nephrotoxicity, ascribed as the most serious toxic effect, has been related to the state of aggregation of the antibiotic. In search of the increase in AmB antifungal activity associated with low toxicity, several AmB-amphiphile formulations have been proposed. This work focuses on the structural characterization of a specific AmB formulation: AmB associated with sonicated dioctadecyl dimethylammonium bromide (DODAB) aggregates. Here, it was confirmed that sonicated DODAB dispersion is constituted by DODAB bicelles, and that monomeric AmB is much more soluble in bicelles than in DODAB vesicles. A new optical parameter is proposed for the estimation of the relative amount of amphiphile-bound monomeric AmB. With theoretical simulations of the spectra of spin labels incorporated in DODAB bicelles it was possible to prove that monomeric AmB binds preferentially to lipids located at the edges of DODAB bicelles, rigidifying them, and decreasing the polarity of the region. That special binding of monomeric AmB along the borders of bicelles, where the lipids are highly disorganized, could be used in the formulation of other carriers for the antibiotic, including mixtures of natural lipids which are known to form bicelles. (C) 2011 Elsevier B.V. All rights reserved.

A Hopping Mechanism for Cargo Transport by Molecular Motors on Crowded Microtubules

Most models designed to study the bidirectional movement of cargos as they are driven by molecular motors rely on the idea that motors of different polarities can be coordinated by external agents if arranged into a motor-cargo complex to perform the necessary work Gross, Hither and yon: a review of bidirectional microtubule-based transport (Gross in Phys. Biol. 1:R1-R11, 2004). Although these models have provided us with important insights into these phenomena, there are still many unanswered questions regarding the mechanisms through which the movement of the complex takes place on crowded microtubules. For example (i) how does cargo-binding affect motor motility? and in connection with that-(ii) how does the presence of other motors (and also other cargos) on the microtubule affect the motility of the motor-cargo complex? We discuss these questions from a different perspective. The movement of a cargo is conceived here as a hopping process resulting from the transference of cargo between neighboring motors. In the light of this, we examine the conditions under which cargo might display bidirectional movement even if directed by motors of a single polarity. The global properties of the model in the long-time regime are obtained by mapping the dynamics of the collection of interacting motors and cargos into an asymmetric simple exclusion process (ASEP) which can be resolved using the matrix ansatz introduced by Derrida (Derrida and Evans in Nonequilibrium Statistical Mechanics in One Dimension, pp. 277-304, 1997; Derrida et al. in J. Phys. A 26: 1493-1517, 1993).