Form III-like conformation and Form I-like packing in a chloroform channel solvate of the diuretic drug chlortalidone

Chlortalidone (CTD) is an antihypertensive drug for which only two solid state phases have been structurally elucidated thus far. Here, we have prepared a chloroform solvate thereof, namely, CTD Form IV, and its structure was compared to those of Form I and Form III. Its two conformers exhibit a dual structural feature in relation to the antecedent polymorphs. Both CTD molecules of Form IV adopt a Form III-like conformation, which is featured, if the conformation of CTD Form I is used as a reference, by a rotation of about 90 degrees on the axis of the C-C bond bridging the substituted benzene and isoindolinyl rings. However, CTD Form IV assembles as in the Form I crystal packing despite the different stacking fashion of their centrosymmetric dimers. In contrast to Form I, there is no offset stacking in Form IV, which forces a bend of ca. 24 degrees between the planes passing through the isoindolinyl moieties of two [100]-stacked dimers. Chloroform molecules at a maximum stoichiometry of 0.25 mol per mol of the drug play a stabilizing role in the assembly of Form IV by filling the channels formed on the crystals.

The regulation of NHE₁ and NHE₃ activity by angiotensin II is mediated by the activation of the angiotensin II type I receptor/phospholipase C/calcium/calmodulin pathway in distal nephron cells

Angiotensin II (Ang II), acting via the AT1 receptor, induces an increase in intracellular calcium [Ca(2+)]i that then interacts with calmodulin (CaM). The Ca(2+)/CaM complex directly or indirectly activates sodium hydrogen exchanger 1 (NHE1) and phosphorylates calmodulin kinase II (CaMKII), which then regulates sodium hydrogen exchanger 3 (NHE3) activity. In this study, we investigated the cellular signaling pathways responsible for Ang II-mediated regulation of NHE1 and NHE3 in Madin-Darby canine kidney (MDCK) cells. The NHE1- and NHE3-dependent pHi recovery rates were evaluated by fluorescence microscopy using the fluorescent probe BCECF/AM, messenger RNA was evaluated with the reverse transcription polymerase chain reaction (RT-PCR), and protein expression was evaluated by immunoblot. We demonstrated that treatment with Ang II (1pM or 1 nM) for 30 min induced, via the AT1 but not the AT2 receptor, an equal increase in NHE1 and NHE3 activity that was reduced by the specific inhibitors HOE 694 and S3226, respectively. Ang II (1 nM) did not change the total expression of NHE1, NHE3 or calmodulin, but it induced CaMKII, cRaf-1, Erk1/2 and p90(RSK) phosphorylation. The stimulatory effects of Ang II (1 nM) on NHE1 or NHE3 activity or protein abundance was reduced by ophiobolin-A (CaM inhibitor), KN93 (CaMKII inhibitor) or PD98059 (Mek inhibitor). These results indicate that after 30 min, Ang II treatment may activate G protein-dependent pathways, including the AT1/PLC/Ca(2+)/CaM pathway, which induces CaMKII phosphorylation to stimulate NHE3 and induces cRaf-1/Mek/Erk1/2/p90(RSK) activity to stimulate NHE1

Influence of the network modifier on the characteristics of MSnO3 (M=Sr and Ca) thin films synthesized by chemical solution deposition

CaSnO3 and SrSnO3 alkaline earth stannate thin films were prepared by chemical solution deposition using the polymeric precursor method on various single crystal substrates (R- and C-sapphire and 100-SrTiO3) at different temperatures. The films were characterized by X-ray diffraction (θ-2θ, ω- and φ-scans), field emission scanning electron microscopy, atomic force microscopy, micro-Raman spectroscopy and photoluminescence. Epitaxial SrSnO3 and CaSnO3 thin films were obtained on SrTiO3 with a high crystalline quality. The long-range symmetry promoted a short-range disorder which led to photoluminescence in the epitaxial films. In contrast, the films deposited on sapphire exhibited a random polycrystalline growth with no meaningful emission regardless of the substrate orientation. The network modifier (Ca or Sr) and the substrate (sapphire or SrTiO3) influenced the crystallization process and/or the microstructure. Higher is the tilts of the SnO6 octahedra, as in CaSnO3, higher is the crystallization temperature, which changed also the nucleation/grain growth process.

Taxonomy of ""Mouse-colored Tapaculos"". I. On the application of the name Malacorhynchus speluncae Menetries, 1835 (Aves: Passeriformes: Rhinocryptidae)

The type specimen of Malacorhynchus speluncae was described and illustrated as being ""mouse gray with a bluish luster"" on the upperparts and as having a ""lighter color on the lower side of the body"" which ""becomes whitish towards the middle of the throat and breast"". It represents a taxon presently placed in the genus Scytalopus. Since 1907, the name Scytalopus speluncae has been attributed to the predominantly dark-gray species from the southeastern coastal Brazilian mountains. Recently, it was suggested that the name S. speluncae should be applied to a species that is light-gray with whitish belly and extensive barring on the flanks and that occurs predominantly in the Espinhaco Range, Minas Gerais state, to the west of the range of the dark-gray taxon. As a consequence, the dark-gray species, presumably without any available name, was described as a new species, S. notorius. However, on the basis of a critical analysis of the available information on the type specimen of S. speluncae, including the original description and illustration (Menetries 1835), and our examination of large series of museum specimens, we demonstrate that the type of S. speluncae falls within the known plumage variation of the dark-gray species and that it does not show the diagnostic characters of the light-gray form. Thus, we propose that the name S. speluncae be applied only to the dark-gray species. Consequently, S. notorius must be regarded a junior-synonym of S. speluncae. Because of problems related to the exact collecting sites of Menetries, and taking into consideration the distribution of the dark-gray species, we suggest ""Serra dos Orgaos"", in Rio de Janeiro state, as the type-locality of S. speluncae.

Mechanical loading induces the expression of a Pol I regulon at the onset of skeletal muscle hypertrophy

von Walden F, Casagrande V, Ostlund Farrants AK, Nader GA. Mechanical loading induces the expression of a Pol I regulon at the onset of skeletal muscle hypertrophy. Am J Physiol Cell Physiol 302: C1523-C1530, 2012. First published March 7, 2012; doi:10.1152/ajpcell.00460.2011.-The main goal of the present study was to investigate the regulation of ribosomal DNA (rDNA) gene transcription at the onset of skeletal muscle hypertrophy. Mice were subjected to functional overload of the plantaris by bilateral removal of the synergist muscles. Mechanical loading resulted in muscle hypertrophy with an increase in rRNA content. rDNA transcription, as determined by 45S pre-rRNA abundance, paralleled the increase in rRNA content and was consistent with the onset of the hypertrophic response. Increased transcription and protein expression of c-Myc and its downstream polymerase I (Pol I) regulon (POL1RB, TIF-1A, PAF53, TTF1, TAF1C) was also consistent with the increase in rRNA. Similarly, factors involved in rDNA transcription, such as the upstream binding factor and the Williams syndrome transcription factor, were induced by mechanical loading in a corresponding temporal fashion. Chromatin immunoprecipitation revealed that these factors, together with Pol I, were enriched at the rDNA promoter. This, in addition to an increase in histone H3 lysine 9 acetylation, demonstrates that mechanical loading regulates rRNA synthesis by inducing a gene expression program consisting of a Pol I regulon, together with accessory factors involved in transcription and chromatin remodeling at the rDNA promoter. Altogether, these data indicate that transcriptional and epigenetic mechanisms take place in the regulation of ribosome production at the onset of muscle hypertrophy.

Petrology and Sr-Nd characteristics of the Nova Lacerda dike swarm, SW Amazonian Craton: new insights regarding its subcontinental mantle source and Mesoproterozoic geodynamics

The NNW-trending Nova Lacerda tholeiitic dike swarm in Mato Grosso State, Central Brazil, intrudes the Nova Lacerda granite (1.46 Ga) and the Jauru granite-greenstone terrain (ca. 1.79-1.77 Ga). The swarm comprises diabases I and II and amphibolites emplaced at ca. 1.38 Ga. Geochemical data indicate that these are evolved tholeiites characterized by high LILE/HSFE and LREE/HSFE ratios. Isotopic modelling yields positive epsilon(Nd)(T) values (+0.86 to +2.65), whereas values for epsilon(Sr)(T) range from positive to negative (+1.96 to -5.56). Crustal contamination did not play a significant petrogenetic role, as indicated by a comparison of isotopic data (Sr-Nd) from both dikes and country rocks, and by the relationship between isotopic and geochemical parameters (SiO2, K2O, Rb/Sr, and La/Yb) of the dikes. We attribute the origin of these tholeiites to fractional crystallization of evolved melts derived from a heterogeneous mantle source. Comparison of the geochemical and isotopic data of the studied swarm and other tholeiitic Mesoproterozoic mafic intrusions of the SWAmazonian Craton the Serra da Providencia, Colorado, and Nova Brasilandia bimodal suites - indicates that parental melts of the Nova Lacerda swarm were derived from the most enriched mantle source. This enrichment was probably caused by the stronger influence of the EMI component on the DMM end-member. These data, coupled with trace element bulk-rock geochemistry of the country rocks, and comparisons with the Colorado Complex of similar age, suggest a continental-margin arc setting for the emplacement of the Nova Lacerda dikes.

Milky Way demographics with the VVV survey I. The 84-million star colour-magnitude diagram of the Galactic bulge

Context. The Milky Way (MW) bulge is a fundamental Galactic component for understanding the formation and evolution of galaxies, in particular our own. The ESO Public Survey VISTA Variables in the Via Lactea is a deep near-IR survey mapping the Galactic bulge and southern plane. Particularly for the bulge area, VVV is covering similar to 315 deg(2). Data taken during 2010 and 2011 covered the entire bulge area in the JHKs bands. Aims. We used VVV data for the whole bulge area as a single and homogeneous data set to build for the first time a single colour-magnitude diagram (CMD) for the entire Galactic bulge. Methods. Photometric data in the JHK(s) bands were combined to produce a single and huge data set containing 173 150 467 sources in the three bands, for the similar to 315 deg(2) covered by VVV in the bulge. Selecting only the data points flagged as stellar, the total number of sources is 84 095 284. Results. We built the largest colour-magnitude diagrams published up to date, containing 173.1+ million sources for all data points, and more than 84.0 million sources accounting for the stellar sources only. The CMD has a complex shape, mostly owing to the complexity of the stellar population and the effects of extinction and reddening towards the Galactic centre. The red clump (RC) giants are seen double in magnitude at b similar to -8 degrees-10 degrees, while in the inner part (b similar to -3 degrees) they appear to be spreading in colour, or even splitting into a secondary peak. Stellar population models show the predominance of main-sequence and giant stars. The analysis of the outermost bulge area reveals a well-defined sequence of late K and M dwarfs, seen at (J - K-s) similar to 0.7-0.9 mag and K-s greater than or similar to 14 mag. Conclusions. The interpretation of the CMD yields important information about the MW bulge, showing the fingerprint of its structure and content. We report a well-defined red dwarf sequence in the outermost bulge, which is important for the planetary transit searches of VVV. The double RC in magnitude seen in the outer bulge is the signature of the X-shaped MW bulge, while the spreading of the RC in colour, and even its splitting into a secondary peak, are caused by reddening effects. The region around the Galactic centre is harder to interpret because it is strongly affected by reddening and extinction.

CD44(+)/CD24(-) cells and lymph node metastasis in stage I and II invasive ductal carcinoma of the breast

The presence of tumor-initiating cells (CD44(+)/CD24(-)) in solid tumors has been reported as a possible cause of cancer metastasis and treatment failure. Nevertheless, little is know about the presence of CD44(+)/CD24(-) cells within the primary tumor and metastasis. The proportion of CD44(+)/CD24(-) cells was analyzed in 40 samples and in 10 lymph node metastases using flow cytometry phenotyping. Anti-human CD326 (EpCam; FITC), antihuman CD227 (MUC-1; FITC), anti-human CD44 (APC), and anti-human CD24 (PE), anti-ABCG2 (PE), and anti-CXCR4 (PeCy7) were used for phenotype analysis. The mean patient age was 60.5 years (range, 33-87 years); mean primary tumor size (pT) was 1.8 cm (0.5-3.5 cm). The Wilcoxon or Kruskal-Wallis test was used for univariate analyses. Logistic regression was used for multivariate analysis. The median percentage of CD44(+)/CD24(-) cells within primary invasive ductal carcinomas (IDC) was 2.7% (range, 0.2-71.2). In lymph node metastases, we observed a mean of 6.1% (range, 0.07-53.7). The percentage of CD44(+)/CD24(-) cells in IDCs was not associated with age, pT, tumor grade and HER2. We observed a significantly enrichment of CD44(+)/CD24(-) and ABCG2(+) cells in ESA(+) cell population in patients with positive lymph nodes (P = 0.02 and P = 0.04, respectively). Our data suggest that metastatic dissemination is associated with an increase in tumorinitiating cells in stage I and II breast cancer.

Synthesis and characterization of [Ru(eta(6)-C10H14)(dppf)X][PF6] (X = Cl, Br, I, SnF3) compounds: The X-ray structure of [Ru(eta(6)-C10H14)(dppf)Cl][SnCl3]center dot 0.45CH(2)Cl(2)

The arene-ruthenium complex [Ru(eta(6)-C10H14)(dppf)Cl]PF6 (1) was used as a precursor for the syntheses of the [Ru(eta(6)-C10H14)(dppf)Br]PF6 (2), [Ru(eta(6)-C10H14)(dppf)I]PF6 (3). [Ru(eta(6)-C10H14)(dppf)SnF3]PF6 (4) and [Ru(eta(6)-C10H14)(dppf)Cl][SnCl3]center dot 0.45CH(2)Cl(2) (5) complexes by its reactions with KBr, Kl, SnF2 and SnCl2. respectively. All of the compounds were characterized by NMR, IR, Fe-57 and Sn-119-Mossbauer spectroscopy, and cyclic voltammetry. The single-crystal X-ray structure analysis of the [Ru(eta(6)-C10H14)(dppf)Cl] [SnCl3]center dot 0.45CH(2)Cl(2) complex revealed the expected piano-stool geometry. Cyclic voltammograms of the complexes showed only one quasi-reversible electrochemical process, involving the oxidation of Fe(II) and Ru(II) at the same potential, which was confirmed by exhaustive electrolysis experiments. Fe-57-Mossbauer parameters obtained for the complexes (1-5) were fitted with one doublet corresponding to a site of one iron(II). The Sn-119-Mossbauer parameters of the complex (4) indicate that tin is tetra covalent. (c) 2012 Elsevier Ltd. All rights reserved.

The Banhado Alkaline Complex, Southeastern Brazil: source and evolution of potassic SiO2-undersaturated high-Ca and low-Ca magmatic series

The Cretaceous Banhado alkaline complex in southeastern Brazil presents two potassic SiO2-undersaturated series. The high-Ca magmatic series consist of initially fractionated olivine (Fo(92-91)) + diopside (Wo(48-43)En(49-35)Ae(0-7)), as evidenced by the presence of xenocrysts and xenoliths. In that sequence, diopside (Wo(47-38)En(46-37)Ae(0-8)) + phlogopite + apatite + perovskite (Prv(> 92)) crystallized to form the phlogopite melteigite and led to the Ca enrichment of the magma. Diopside (Wo(47-41)En(32-24) Ae(3-14)) continued to crystallize as an early mafic mineral, followed by nepheline (Ne(74.8-70.1)Ks(26.3-21.2)Qz(7.6-0.9)) and leucite (Lc(65-56)) and subsequently by melanite and potassic feldspar (Or(85-99)Ab(1-7)) to form melanite ijolites, wollastonite-melanite urtites and melanite-nepheline syenites. Melanite-pseudoleucite-nepheline syenites are interpreted to be a leucite accumulation. Melanite nephelinite dykes are believed to represent some of the magmatic differentiation steps. The low-Ca magmatic series is representative of a typical fractionation of aegirine-augite (Wo(36-29)En(25-4)Ae(39-18)) + alkali feldspar (Or(57-96)Ab(3-43)) + nepheline (Ne(76.5-69.0)Ks(19.9-14.4)Qz(15.1-7.7)) + titanite from phonolite magma. The evolution of this series from potassic nepheline syenites to sodic sodalite syenites and sodalitolites is attributed to an extensive fractionation of potassic feldspar, which led to an increase of the NaCl activity in the melt during the final stages forming sodalite-rich rocks. Phonolite dykes followed a similar evolutionary process and also registered some crustal assimilation. The mesocratic nepheline syenites showed interactions with phlogopite melteigites, such as compatible trace element enrichments and the presence of diopside xenocrysts, which were interpreted to be due to a mixing/mingling process of phonolite and nephelinite magmas. The geochemical data show higher TiO2 and P2O5 contents and lower SiO2 contents for the high-Ca series and different LILE evolution trends and REE chondrite-normalized patterns as compared to the low-Ca series. The Sr-87/Sr-86, Nd-143/Nd-144, Pb-206/Pb-204 and Pb-208/Pb-204 initial ratios for the high-Ca series (0.70407-0.70526, 0.51242-0.51251, 17.782-19.266 and 38.051-39.521, respectively) were slightly different from those of the low-Ca series (0.70542-0.70583, 0.51232-0.51240, 17.758-17.772 and 38.021-38.061, respectively). For both series, a CO2-rich potassic metasomatized lithospheric mantle enriched the source with rutile-bearing phlogopite clinopyroxenite veins. Kamafugite-like parental magma is attributed to the high-Ca series with major contributions from the melting of the veins. Potassic nephelinite-like parental magma is assigned to the low-Ca series, where the metasomatized wall-rock played a more significant role in the melting process.

Biomimetic in vitro oxidation of lapachol: A model to predict and analyse the in vivo phase I metabolism of bioactive compounds

The bioactive naphtoquinone lapachol was studied in vitro by a biomimetic model with Jacobsen catalyst (manganese(III) salen) and iodosylbenzene as oxidizing agent. Eleven oxidation derivatives were thus identified and two competitive oxidation pathways postulated. Similar to Mn(III) porphyrins, Jacobsen catalyst mainly induced the formation of para-naphtoquinone derivatives of lapachol, but also of two ortho-derivatives. The oxidation products were used to develop a GC MS (SIM mode) method for the identification of potential phase I metabolites in vivo. Plasma analysis of Wistar rats orally administered with lapachol revealed two metabolites, alpha-lapachone and dehydro-alpha-lapachone. Hence, the biomimetic model with a manganese salen complex has evidenced its use as a valuable tool to predict and elucidate the in vivo phase I metabolism of lapachol and possibly also of other bioactive natural compounds. (C) 2012 Elsevier Masson SAS. All rights reserved.

Comparative analysis of the anterior and posterior length and deflection angle of the cranial base, in individuals with facial pattern I, II and III

Objective: This study evaluated the variations in the anterior cranial base (S-N), posterior cranial base (S-Ba) and deflection of the cranial base (SNBa) among three different facial patterns (Pattern I, II and III). Method: A sample of 60 lateral cephalometric radiographs of Brazilian Caucasian patients, both genders, between 8 and 17 years of age was selected. The sample was divided into 3 groups (Pattern I, II and III) of 20 individuals each. The inclusion criteria for each group were the ANB angle, Wits appraisal and the facial profile angle (G’.Sn.Pg’). To compare the mean values obtained from (SNBa, S-N, S-Ba) each group measures, the ANOVA test and Scheffé’s Post-Hoc test were applied. Results and Conclusions: There was no statistically significant difference for the deflection angle of the cranial base among the different facial patterns (Patterns I, II and III). There was no significant difference for the measures of the anterior and posterior cranial base between the facial Patterns I and II. The mean values for S-Ba were lower in facial Pattern III with statistically significant difference. The mean values of S-N in the facial Pattern III were also reduced, but without showing statistically significant difference. This trend of lower values in the cranial base measurements would explain the maxillary deficiency and/or mandibular prognathism features that characterize the facial Pattern III.

Identification of SL addition trans-splicing acceptor sites in the internal transcribed spacer I region of pre-rRNA in Leishmania (Leishmania) amazonensis

Trypanosomatidae is a family of early branching eukaryotes harbouring a distinctive repertoire of gene expression strategies. Functional mature messenger RNA is generated via the trans-splicing and polyadenylation processing of constitutively transcribed polycistronic units. Recently, trans-splicing of pre-small subunit ribosomal RNA in the 5' external transcribed spacer region and of precursor tRNAsec have been described. Here, we used a previously validated semi-nested reverse transcription-polymerase chain reaction strategy to investigate internal transcribed spacer (ITS) I acceptor sites in total RNA from Leishmania (Leishmania) amazonensis. Two distinct spliced leader-containing RNAs were detected indicating that trans-splicing reactions occur at two AG acceptor sites mapped in this ITS region. These data provide further evidence of the wide spectrum of RNA molecules that act as trans-splicing acceptors in trypanosomatids.

Study of the number of occlusal contacts in maximum intercuspation before orthodontic treatment in subjects with Angle Class I and Class II Division 1 malocclusion

OBJECTIVE: Define and compare numbers and types of occlusal contacts in maximum intercuspation. METHODS: The study consisted of clinical and photographic analysis of occlusal contacts in maximum intercuspation. Twenty-six Caucasian Brazilian subjects were selected before orthodontic treatment, 20 males and 6 females, with ages ranging between 12 and 18 years. The subjects were diagnosed and grouped as follows: 13 with Angle Class I malocclusion and 13 with Angle Class II Division 1 malocclusion. After analysis, the occlusal contacts were classified according to the established criteria as: tripodism, bipodism, monopodism (respectively, three, two or one contact point with the slope of the fossa); cuspid to a marginal ridge; cuspid to two marginal ridges; cuspid tip to opposite inclined plane; surface to surface; and edge to edge. RESULTS: The mean number of occlusal contacts per subject in Class I malocclusion was 43.38 and for Class II Division 1 malocclusion it was 44.38, this difference was not statistically significant (p>0.05). CONCLUSIONS: There is a variety of factors that influence the number of occlusal contacts between a Class I and a Class II, Division 1 malocclusion. There is no standardization of occlusal contact type according to the studied malocclusions. A proper selection of occlusal contact types such as cuspid to fossa or cuspid to marginal ridge and its location in the teeth should be individually defined according to the demands of each case. The existence of an adequate occlusal contact leads to a correct distribution of forces, promoting periodontal health.