Quantitative analysis of mannitol polymorphs: X-ray powder diffractometry—exploring preferred orientation effects

Mannitol is a polymorphic pharmaceutical excipient, which commonly exists in three forms: alpha, beta and delta. Each polymorph has a needle-like morphology, which can give preferred orientation effects when analysed by X-ray powder diffractometry (XRPD) thus providing difficulties for quantitative XRPD assessments. The occurrence of preferred orientation may be demonstrated by sample rotation and the consequent effects on X-ray data can be minimised by reducing the particle size. Using two particle size ranges (less than 125 and 125–500�microns), binary mixtures of beta and delta mannitol were prepared and the delta component was quantified. Samples were assayed in either a static or rotating sampling accessory. Rotation and reducing the particle size range to less than�125 microns halved the limits of detection and quantitation to 1 and 3.6%, respectively. Numerous potential sources of assay errors were investigated; sample packing and mixing errors contributed the greatest source of variation. However, the rotation of samples for both particle size ranges reduced the majority of assay errors examined. This study shows that coupling sample rotation with a particle size reduction minimises preferred orientation effects on assay accuracy, allowing discrimination of two very similar polymorphs at around the 1% level

Lipoma preferred partner is a mechanosensitive protein regulated by nitric oxide in the heart

Adaptor proteins play an important role in signaling pathways by providing a platform on which many other proteins can interact. Malfunction or mislocalization of these proteins may play a role in the development of disease. Lipoma preferred partner (LPP) is a nucleocytoplasmic shuttling adaptor protein. Previous work shows that LPP plays a role in the function of smooth muscle cells and in atherosclerosis. In this study we wanted to determine whether LPP has a role in the myocardium. LPP expression increased by 56% in hearts from pressure overload aortic-banded rats (p < 0.05 n = 4), but not after myocardial infarction, suggesting hemodynamic load regulates its expression. In vitro, LPP expression was 87% higher in cardiac fibroblasts than myocytes (p < 0.05 n = 3). LPP expression was downregulated in the absence of the actin cytoskeleton but not when microtubules were disassembled. We mechanically stretched cardiac fibroblasts using the Flexcell 4000 for 48 h (1 Hz, 5% maximum strain), which decreased total LPP total expression and membrane localization in subcellular fractions (p < 0.05, n = 5). However, L-NAME, an inhibitor of nitric oxide synthase (NOS), significantly upregulated LPP expression. These findings suggest that LPP is regulated by a complex interplay between NO and mechanical cues and may play a role in heart failure induced by increased hemodynamic load.

Preferred orientation in an angled intercalation site of a chloro-substituted lambda- [Ru(TAP)2(dppz)]2+ complex bound to d(TCGGCGCCGA)2

The crystal structure of the ruthenium DNA ‘light-switch’ complex -[Ru(TAP)2(11-Cl-dppz)]2+ (TAP = tetraazaphenanthrene, dppz = dipyrido[3,2-a':2',3'-c]phenazine)) bound to the oligonucleotide duplex d(TCGGCGCCGA)2 is reported. The synthesis of the racemic ruthenium complex is described for the first time, and the racemate was used in this study. The crystal structure, at atomic resolution (1.0 Å), shows one ligand as a wedge in the minor groove, resulting in the 51 kinking of the double helix, as with the parent lambda-[Ru(TAP)2(dppz)]2+. Each complex binds to one duplex by intercalation of the dppz ligand and also by semi-intercalation of one of the orthogonal TAP ligands into a second symmetrically equivalent duplex. The 11-Cl substituent binds with the major component (66%) oriented with the 11-chloro substituent on the purine side of the terminal step of the duplex.

Modulation of stretch-induced myocyte remodeling and gene expression by nitric oxide: a novel role for lipoma preferred partner in myofibrillogenesis

Prolonged hemodynamic load as a result of hypertension eventually leads to maladaptive cardiac adaptation and heart failure. The signalling pathways that underlie these changes are still poorly understood. The adaptive response to mechanical load is mediated by mechanosensors which convert the mechanical stimuli into a biological response. We examined the effect of cyclic mechanical stretch on myocyte adaptation using neonatal rat ventricular myocytes with 10% (adaptive) or 20% (maladaptive) maximum strain, 1Hz for 48 hours to mimic in vivo mechanical stress. Cells were also treated with and without L-NAME, a general nitric oxide synthase (NOS) inhibitor to suppress NO production. Maladaptive 20% mechanical stretch led to a significant loss of intact sarcomeres which was rescued by LNAME (P<0.05, n≥5 cultures). We hypothesized that the mechanism was through NOinduced alteration of myocyte gene expression. L-NAME up-regulated the mechanosensing proteins Muscle LIM protein (MLP (by 100%, p<0.05, n=4 cultures)) and lipoma preferred partner, a novel cardiac protein (LPP (by 80%, p<0.05, n=4 cultures)). L-NAME also significantly altered the subcellular localisation of LPP and MLP in a manner that favoured growth and adaptation. These findings suggest that NO participates in stretch-mediated adaptation. The use of isoform selective NOS inhibitors indicated a complex interaction between iNOS and nNOS isoforms regulate gene expression. LPP knockdown by siRNA led to formation of α-actinin aggregates and Z-bodies showing that myofibrillogenesis was impaired. There was an up-regulation of E3 ubiquitin ligase (MUL1) by 75% (P<0.05, n=5 cultures). This indicates that NO contributes to stretch-mediated adaptation via the upregulation of proteins associated mechansensing and myofibrillogenesis, thereby presenting potential therapeutic targets during the progression of heart failure. Keywords: Mechanotransduction, heart failure, stretch, heart, hypertrophy

Determination of orientations of aromatic groups in self-assembled peptide fibrils by polarised Raman spectroscopy

In this paper we describe a novel combination of Raman spectroscopy, isotope editing and X-ray scattering as a powerful approach to give detailed structural information on aromatic side chains in peptide fibrils. The orientation of the tyrosine residues in fibrils of the peptide YTIAALLSPYS with respect to the fibril axis has been determined from a combination of polarised Raman spectroscopy and X-ray diffraction measurements. The Raman intensity of selected tyrosine bands collected at different polarisation geometries is related to the values and orientation of the Raman tensor for those specific vibrations. Using published Raman tensor values we solved the relevant expressions for both of the two tyrosine residues present in this peptide. Ring deuteration in one of the two tyrosine side chains allowed for the calculation to be performed individually for both, by virtue of the isotopic shift that eliminates band overlapping. Sample disorder was taken into account by obtaining the distribution of orientations of the samples from X-ray diffraction experiments. The results provide previously unavailable details about the molecular conformation of this peptide, and demonstrate the value of this approach for the study of amyloid fibrils.

X‑ray crystal structure of rac-[Ru(phen)2dppz]2+ with d(ATGCAT)2 shows enantiomer orientations and water ordering

We report an atomic resolution X-ray crystal structure containing both enantiomers of rac-[Ru(phen)2dppz]2+ with the d-(ATGCAT)2 DNA duplex (phen = phenanthroline; dppz = dipyridophenazine). The first example of any enantiomeric pair crystallized with a DNA duplex shows different orientations of the Λ and Δ binding sites, separated by a clearly defined structured water monolayer. Job plots show that the same species is present in solution. Each enantiomer is bound at a TG/CA step and shows intercalation from the minor groove. One water molecule is directly located on one phenazine N atom in the Δ-enantiomer only.

Preferred response of the East Asian summer monsoon to local and non-local anthropogenic sulphur dioxide emissions

In this study, the atmospheric component of a state-of-the-art climate model (HadGEM2-ES) that includes earth system components such as interactive chemistry and eight species of tropospheric aerosols considering aerosol direct, indirect, and semi-direct effects, has been used to investigate the impacts of local and non-local emissions of anthropogenic sulphur dioxide on the East Asian summer monsoon (EASM). The study focuses on the fast responses (including land surface feedbacks, but without sea surface temperature feedbacks) to sudden changes in emissions from Asia and Europe. The initial responses, over days 1–40, to Asian and European emissions show large differences. The response to Asian emissions involves a direct impact on the sulphate burden over Asia, with immediate consequences for the shortwave energy budget through aerosol–radiation and aerosol–cloud interactions. These changes lead to cooling of East Asia and a weakening of the EASM. In contrast, European emissions have no significant impact on the sulphate burden over Asia, but they induce mid-tropospheric cooling and drying over the European sector. Subsequently, however, this cold and dry anomaly is advected into Asia, where it induces atmospheric and surface feedbacks over Asia and the Western North Pacific (WNP), which also weaken the EASM. In spite of very different perturbations to the local aerosol burden in response to Asian and European sulphur dioxide emissions, the large scale pattern of changes in land–sea thermal contrast, atmospheric circulation and local precipitation over East Asia from days 40 onward exhibits similar structures, indicating a preferred response, and suggesting that emissions from both regions likely contributed to the observed weakening of the EASM. Cooling and drying of the troposphere over Asia, together with warming and moistening over the WNP, reduces the land–sea thermal contrast between the Asian continent and surrounding oceans. This leads to high sea level pressure (SLP) anomalies over Asia and low SLP anomalies over the WNP, associated with a weakened EASM. In response to emissions from both regions warming and moistening over the WNP plays an important role and determines the time scale of the response.

Comparing detection and location performance of perpendicular and parallel broadside GPR antenna orientations

GPR (Ground Penetrating Radar) results are shown for perpendicular broadside and parallel broadside antenna orientations. Performance in detection and localization of concrete tubes and steel tanks is compared as a function of acquisition configuration. The comparison is done using 100 MHz and 200 MHz center frequency antennas. All tubes and tanks are buried at the geophysical test site of IAG/USP in Sao Paulo city, Brazil. The results show that the long steel pipe with a 38-mm diameter was well detected with the perpendicular broadside configuration. The concrete tubes were better detected with the parallel broadside configuration, clearly showing hyperbolic diffraction events from all targets up to 2-m depth. Steel tanks were detected with the two configurations. However, the parallel broadside configuration was generated to a much lesser extent an apparent hyperbolic reflection corresponding to constructive interference of diffraction hyperbolas of adjacent targets placed at the same depth. Vertical concrete tubes and steel tanks were better contained with parallel broadside antennas, where the apexes of the diffraction hyperbolas better corresponded to the horizontal location of the buried target disposition. The two configurations provide details about buried targets emphasizing how GPR multi-component configurations have the potential to improve the subsurface image quality as well as to discriminate different buried targets. It is judged that they hold some applicability in geotechnical and geoscientific studies. (C) 2009 Elsevier B.V. All rights reserved.

Intermolecular Contacts Influencing the Conformational and Geometric Features of the Pharmaceutically Preferred Mebendazole Polymorph C

Mebendazole (MBZ) is a common benzimidazole anthelmintic that exists in three different polymorphic forms, A, B, and C. Polymorph C is the pharmaceutically preferred form due to its adequated aqueous solubility. No single crystal structure determinations depicting the nature of the crystal packing and molecular conformation and geometry have been performed on this compound. The crystal structure of mebendazole form C is resolved for the first time. Mebendazole form C crystallizes in the triclinic centrosymmetric space group and this drug is practically planar, since the least-squares methyl benzimidazolylcarbamate plane is much fitted on the forming atoms. However, the benzoyl group is twisted by 31(1)degrees from the benzimidazole ring, likewise the torsional angle between the benzene and carbonyl moieties is 27(1)degrees. The formerly described bends and other interesting intramolecular geometry features were viewed as consequence of the intermolecular contacts occurring within mebendazole C structure. Among these features, a conjugation decreasing through the imine nitrogen atom of the benzimidazole core and a further resonance path crossing the carbamate one were described. At last, the X-ray powder diffractogram of a form C rich mebendazole mixture was overlaid to the calculated one with the mebendazole crystal structure. (C) 2008 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 98:2336-2344, 2009

Approach to culture through value orientations

What existential premises, what artifacts are walked through to make it possible for a person to step assuredly in reality? What scope exists for the frame shape? Are there doorways all peoples have to frame, and is there a limited stock to choose from? These were questions motivating the research which this paper reports.

Why did democracy become the preferred political regime only in the twentieth century?

A Democracia Tornou-Se a Forma Preferida de Governo Apenas no Século Vinte. a Busca por Motivos Racionais para Entendermos a Razão não é Suficiente. o Autor Procura por um Fato Histórico Novo que Tenha Levado a Essa Mudança de Preferência, Baseado Sucessivamente na Revolução Capitalista e na Perda Gradual do Medo da Expropriação Pela Burguesia. a Revolução Capitalista que Mudou o Modo de Apropriação do Excedente, da Violência para o Mercado, é a Primeira Condição Necessária. Representa Também a Transição do Estado Absoluto para o Estado Liberal. a Segunda Condição é o Desaparecimento do Medo da Expropriação, Permitindo a Transição do Regime Liberal para o Regime Liberal-Democrático. Depois de Estabelecer Estas Duas Condições, ou Estes Dois Fatos Históricos Novos, o Regime Democrático Tornou-Se Rational Choice para Todas as Classes. para os Capitalistas, Democracia é Agora a Forma de Governo que Melhor Assegura o Direito À Propriedade e Aos Contratos. para os Trabalhadores, é a Forma de Governo que Melhor Assegura o Aumento de Salários com Lucros

A relação gramatical objeto direto : implicações para o ensino de língua materna

This paper attempts to investigate the discourse manifestations of the grammatical relation direct object with respect to the syntactic, semantic and pragmatic properties that underlie this element. The research adopts theoretical orientation of the functionalism from North American and Brazilian schools inspired in Givón (1995, 2001), Hopper and Thompson (1980), Chafe (1979), Furtado da Cunha, Oliveira, Martelotta (2003) inter alia. From functionalism, the research uses principles of iconicity, markedness and informativity and it analize categories of transitivity, grounding and animacy. This research is anchored in prototype model (TAYLOR 1995); construction grammar model (GOLDBERG 1996, 2002). Both theoretical orientations share the view that language is a malleable living organism subject to socio-cultural context. Grammar is then the result of created, maintained, and systematized linguistic patterns developed from and used for language use. According to a functional linguistics and cognitivist linguistics verbs are stored in the speakers lexicon in syntactic-semantic frames which are more frequent. These frames carry information concerning obligatory and optional arguments and the semantic roles these arguments take in the clause. The analysis focuses on the semantic type of the verbs and its relationship with the argument encoded as a direct object observing the aspectual nature of verbs. Direct objects are classified according to their morphology (lexical or pronominal noun phrase), semantic role, informational content and animacy. This study discusses pedagogical implications with relation to how the grammatical concepts touched on this paper are treated in school textbooks. The empirical data come from Corpus Discurso & Gramática: a língua falada e escrita na cidade do Natal (FURTADO DA CUNHA, 1998). This corpus is composed of texts that contain spoken and written modalities. These modalities are in turn organized according to different types: personal narratives, retold narrative, description of preferred place, procedural place, procedural description and report on argumentation. The sample data totals 40 texts produced by four language consultants of the last graduation date. The paper shows that the same syntactic structures (formed through Subject-Verb-Object) correspond to different semantic-pragmatic structures in relation to specific communicative purposes even verb is an event, process or state. The argument structure are not aleatory but are related to experience; that is the way humans conceptualize the world and talk about it

EFFECTS of PREFERRED and NONPREFERRED MUSIC on CONTINUOUS CYCLING EXERCISE PERFORMANCE

The aim of this study was to investigate the effects of preferred and nonpreferred music on exercise distance, Heart Rate (HR), and Rating of Perceived Exertion (RPE) during continuous cycling exercise performed at high intensity Fifteen participants performed five test sessions During two sessions, they cycled with fixed workload on ergometer to determine the Critical Power (Cl') intensity Then, they performed three more sessions cycling at CP intensity listening to Preferred Music, listening to Nonpreferred Music, and No Music The HR responses in the exercise sessions did not differ among all conditions However, the RPE was higher for Nonpreferred Music than in the other conditions The performance under Preferred Music (9 8 +/- 4 6km) was greater than under Nonpreferred Music (7 1 +/- 3 5km) conditions Therefore, listening to Preferred Music during continuous cycling exercise at high intensity can Increase the exercise distance, and individuals listening to Nonpreferred Music can perceive more discomfort caused by the exercise

Attending to the non-preferred hand improves bimanual coordination in children

The effect of attentional focus in bimanual coordination was investigated from a developmental perspective by examining performance of right- and left-handed children, 5-8-years and 9-12-years old, on bimanual reciprocal tapping tasks. Attentional focus was either specified, by asking the children to attend to the preferred or to the non-preferred hand, or unspecified for the execution of the tasks. When attention was oriented to the non-preferred hand we found a reduced movement time and a lower frequency of errors. Performance differences for handedness and age-groups were observed when the children were oriented to attend to the preferred hand or when there was no instruction regarding attention. These differences in performance were eliminated when attention was oriented to the non-preferred hand. (C) 2004 Elsevier B.V. All rights reserved.