Co-Expression of Leptin and Oestrogen Receptors in the Preoptic-Hypothalamic Area

The interaction between the reproductive axis and energy balance suggests that leptin acts as a possible mediator. This hormone acts in the regulation of metabolism, feeding behaviour and reproduction. Animals homozygous for the gene `ob` (ob/ob) are obese and infertile, and these effects are reversed after systemic administration of leptin. Thus, the present study aimed to determine: (i) whether cells that express leptin also express oestrogen receptors of type-alpha (ER-alpha) or -beta (ER-beta) in the medial preoptic area (MPOA) and in the arcuate (ARC), dorsomedial (DMH) and ventromedial hypothalamic nucleus and (ii) whether there is change in the gene and protein expression of leptin in these brain areas in ovariectomised (OVX) animals when oestrogen-primed. Wistar female rats with normal oestrous cycles or ovariectomised oestrogen-primed or vehicle (oil)-primed were utilised. To determine whether there was a co-expression, immunofluorescence was utilised for double staining. Confocal microscopy was used to confirm the co-expression. The technique of real-time polymerase chain reaction and western blotting were employed to analyse gene and protein expression, respectively. The results obtained showed co-expression of leptin and ER-alpha in the MPOA and in the DMH, as well as leptin and ER-beta in the MPOA, DMH and ARC. However, we did not detect leptin in the MPOA, ARC and DMH using western blotting and there was no statistical difference in leptin gene expression in the MPOA, DMH, ARC, pituitary or adipose tissue between OVX rats treated with oestrogen or vehicle. In conclusion, the results obtained in the present study confirm that the brain is also a source of leptin and reveal co-expression of oestrogen receptors and leptin in the same cells from areas related to reproductive function and feeding behaviour. Although these data corroborate the previous evidence obtained concerning the interaction between the action of brain leptin and reproductive function, the physiological relevance of this interaction remains uncertain and additional studies are necessary to elucidate the exact role of central leptin.

In vitro chemosensitivity of canine mast cell tumors grades II and III to all-trans-retinoic acid (ATRA).

Mast cell tumor (MCT) is one of the most prevalent neoplasms that affect the skin and soft tissue of dogs. Because mast cell tumors present a great variety of clinical appearance and behavior, their treatment becomes a challenge. While retinoids are well recognized as promising antitumor agents, there have been only a few reports about retinoids` effect on canine cancers. The aim of this study was to investigate the chemosensitivity of MCT grades II and III to all-trans retinoic acid (ATRA). Immediately after surgical resection, MCT were prepared for primary culture. Samples of MCTs were also fixed in formalin for histopathology and grading according to the classification of Patnaik et al. (Veterinary Pathology 21(5):469-474, 1984). The best results were obtained when neoplastic mast cells were co-cultivated with fibroblasts. Cultured mast cells were, then, treated with concentrations of 10(-4) to 10(-7) M of ATRA, in order to evaluate their chemosensitivity to this retinoid. MTT assay was performed to estimate cell growth and death. The highest level of mast cell chemosensivity was obtained at the dose of 10(-4) M (p < 0,002). MCT of grades II or III were equally susceptible to the treatment with ATRA. Cell death was observed on the first 24 h until 48 h. According to these results, ATRA may be a potential chemotherapeutic agent for the treatment of canine MCT.

Rehardening of acid-softened enamel and prevention of enamel softening through CO(2) laser irradiation

Objectives: The aims of the present study were to investigate whether irradiation with a CO(2) laser could prevent surface softening (i) in sound and (ii) in already softened enamel in vitro. Methods: 130 human enamel samples were obtained and polished with silicon carbide papers. They were divided into 10 groups (n = 13) receiving 5 different surface treatments: laser irradiation (L), fluoride (AmF/NaF gel) application (F), laser prior to fluoride (LF), fluoride prior to laser (FL), non-treated control (C); and submitted to 2 different procedures: half of the groups was acid-softened before surface treatment and the other half after. Immersion in 1% citric acid was the acid challenge. Surface microhardness (SMH) was measured at baseline, after softening and after treatment. Additionally, fluoride uptake in the enamel was quantified. The data were statistically analysed by two-way repeated measurements ANOVA and post hoc comparisons at 5% significance level. Results: When softening was performed either before or after laser treatment, the L group presented at the end of the experiments SMH means that were not significantly different from baseline (p = 0.8432, p = 0.4620). Treatment after softening resulted for all laser groups in statistically significant increase in SMH means as compared to values after softening (p < 0.0001). Enamel fluoride uptake was significantly higher for combined laser-fluoride treatment than in control (p < 0.0001). Conclusion: Irradiation of dental enamel with a CO(2) laser at 0.3J/cm(2) (5 mu s, 226 Hz) not only significantly decreased erosive mineral loss (97%) but also rehardened previously softened enamel in vitro. (C) 2011 Elsevier Ltd. All rights reserved.

Effects of concentrated viral communities on photosynthesis and community composition of co-occurring benthic microalgae and phytoplankton

Marine viruses have been shown to affect phytoplankton productivity; however, there are no reports on the effect of viruses on benthic microalgae (microphytobenthos). Hence, this study investigated the effects of elevated concentrations of virus-like particles on the photosynthetic physiology and community composition of benthic microalgae and phytoplankton. Virus populations were collected near the sediment surface and concentrated by tangential flow ultrafiltration, and the concentrate was added to benthic and water column samples that were obtained along a eutrophication gradient in the Brisbane River/Moreton Bay estuary, Australia. Photosynthetic and community responses of benthic microalgae, phytoplankton and bacteria were monitored over 7 d in aquaria and in situ. Benthic microalgal communities responded to viral enrichment in both eutrophic and oligotrophic sediments. In eutrophic sediments, Euglenophytes (Euglena sp.) and bacteria decreased in abundance by 20 to 60 and 26 to 66%, respectively, from seawater controls. In oligotrophic sediments, bacteria decreased in abundance by 30 to 42% from seawater controls but the dinoflagellate Gymnodinium sp. increased in abundance by 270 to 3600% from seawater controls, The increased abundance of Gymnodinium sp. may be related to increased availability of dissolved organic matter released from lysed bacteria. Increased (140 to 190% from seawater controls) initial chlorophyll a fluorescence measured with a pulse-amplitude modulated fluorometer was observed in eutrophic benthic microalgal incubations following virus enrichment, consistent with photosystem II damage. Virus enrichment in oligotrophic water significantly stimulated carbon fixation rates, perhaps due to increased nutrient availability by bacterial lysis. The interpretation of data from virus amendment experiments is difficult due to potential interaction with unidentified bioactive compounds within seawater concentrates. However, these results show that viruses are capable of influencing microbial dynamics in sediments.

Synthesis and structural properties of patellamide a derivatives and their copper(II) compounds

The synthesis, characterization and copper(II) coordination chemistry of three new cyclic peptide ligands, PatJ(1) (cyclo-(Ile -Thr- (Gly)Thz-lle-Thr(Gly)Thz)), PatJ(2) (cyclo-(Ile-Thr(Gly)Thz-(D)-Ile-Thr-(Gly)Thz)), and PatL (cyclo-(Ile-Ser-(Gly)Thz-Ile-Ser(Gly)Thz)) are reported. All of these cyclic peptides and PatN (cyclo-(Ile-Ser(Gly)Thz-Ile-Thr-(Gly)Thz)) are derivatives of patellamide A and have a [24]azacrown-8 macrocyclic structure. All four synthetic cyclic peptides have two thiazole rings but, in contrast to patellamide A, no oxazoline rings. The molecular structure of PatJ1, determined by X-ray crystallography, has a saddle conformation with two close-to-co-parallel thiazole rings, very similar to the geometry of patellamide D. The two coordination sites of PatJ1 with thiazole-N and amide-N donors are each well preorganized for transition metal ion binding. The coordination of copper(II) was monitored by UV/Vis spectroscopy, and this reveals various (meta)stable mono- and dinuclear copper(II) complexes whose stoichiometry was confirmed by mass spectra. Two types of dinuclear copper(II) complexes, [Cu-2(H4L)(OH2)(n)](2+) (n = 6, 8) and [Cu-2(H4L)(OH2)(n)] (n=4, 6; L=PatN, PatL, PatJ1, PatJ2) have been identified and analyzed structurally by EPR spectroscopy and a combination of spectra simulations and molecular mechanics calculations (MM-EPR). The four structures are similar to each other and have a saddle conformation, that is, derived from the crystal structure of PatJ(1) by a twist of the two thiozole rings. The small but significant structural differences are characterized by the EPR simulations.

Caveolin interacts with the angiotensin II type 1 receptor during exocytic transport but not at the plasma membrane

The mechanisms involved in angiotensin II type 1 receptor (AT(1)-R) trafficking and membrane localization are largely unknown. In this study, we examined the role of caveolin in these processes. Electron microscopy of plasma membrane sheets shows that the AT(1)-R is not concentrated in caveolae but is clustered in cholesterol-independent microdomains; upon activation, it partially redistributes to lipid rafts. Despite the lack of AT(1)-R in caveolae, AT(1)-R. caveolin complexes are readily detectable in cells co-expressing both proteins. This interaction requires an intact caveolin scaffolding domain because mutant caveolins that lack a functional caveolin scaffolding domain do not interact with AT(1)-R. Expression of an N-terminally truncated caveolin-3, CavDGV, that localizes to lipid bodies, or a point mutant, Cav3-P104L, that accumulates in the Golgi mislocalizes AT(1)-R to lipid bodies and Golgi, respectively. Mislocalization results in aberrant maturation and surface expression of AT(1)-R, effects that are not reversed by supplementing cells with cholesterol. Similarly mutation of aromatic residues in the caveolin-binding site abrogates AT(1)-R cell surface expression. In cells lacking caveolin-1 or caveolin-3, AT(1)-R does not traffic to the cell surface unless caveolin is ectopically expressed. This observation is recapitulated in caveolin-1 null mice that have a 55% reduction in renal AT(1)-R levels compared with controls. Taken together our results indicate that a direct interaction with caveolin is required to traffic the AT(1)-R through the exocytic pathway, but this does not result in AT(1)-R sequestration in caveolae. Caveolin therefore acts as a molecular chaperone rather than a plasma membrane scaffold for AT(1)-R.

Geometric isomers of chloro(6-methyl-1,4,8,11-tetraazacyclotetradecane-6-amine)cobalt(III) tetrachlorozincate(II)

The crystal structures of a pair of cis and trans isomers of the macrocyclic chloropentaamine title complex, as their tetrachlorozincate(II) salts, [CoCl(C11H27N5)][ZnCl4], are reported. The two distinct isomeric forms lead to significant variations in the Co-N bond lengths and, furthermore, hydrogen bonding between the complex ions is influenced by the folded (cis) or planar (trans) conformations of the coordinated ligand.

Avaliação de três cepas de vírus rábico, antigenicamente distintas, em camundongos: II - Estudo da disseminação viral por diferentes órgãos

Estudou-se, comparativamente, o grau de disseminação de três cepas de vírus rábico, duas de origem de cão, Jales e Nigéria, e uma de origem de morcego, DR 19, com perfis antigênicos do nucleocapside distintos. Estas cepas foram inoculadas por via intramuscular, na face interna da coxa, em dois grupos de camundongos, com 21 e 28 dias de idade. Os animais foram mantidos em observação por um período total de 30 dias, e dos animais vitimados pela infecção, foram coletados diferentes órgãos, músculo lingual, coração, pulmão, rim e fígado, além do cérebro e da medula espinal, para avaliar-se o grau de disseminação de cada cepa viral, através da prova de imunofluorescência direta (IFD). Os resultados obtidos evidenciaram que os decalques de cérebro e de medula espinal apresentaram total concordância na prova de IFD, constatando-se as maiores diferenças com as cepas Jales e Nigéria, situando-se a cepa DR 19, intermediariamente, a estas duas. O músculo lingual foi o órgão que apresentou maior freqüência de positividade para ambos os grupos etários e para as três cepas virais.

Síntese de complexos de ferro (II) e Ruténio (II) com potenciais aplicações como condutores moleculares

De sua definição, um condutor molecular é “ uma molécula dimensional” que permite a troca de um electrão entre as extremidades através de uma ponte, sendo elas mesmas capazes de trocar electrões com o exterior. Os condutores moleculares envolvendo centros metálicos podem ser constituídos por um sistema orgânico deslocalizado que estabelece a ponte entre os centros metálicos terminais, sendo ambas as partes responsáveis pelo desempenho final do condutor molecular. Os espaçadores orgânicos são responsáveis pelas propriedades electrónicas e pelo comprimento do condutor, enquanto que os centros metálicos, ao introduzirem electrões no sistema, aumentam a condutividade. No capítulo 1 faz-se uma breve introdução aos condutores moleculares, como podem ser avaliados segundo a sua estrutura. Outro assunto abordado é a influência de compostos, mostrando deste modo a interferência do tipo de espaçadores utilizados. Faz-se ainda uma breve apresentação sobre os compostos homo e heterobimetálicos. Finalmente, apresenta-se uma pequena revisão sobre os compostos organometálicos σ-metálicos. No capítulo 2 apresenta-se a síntese e caracterização dos compostos orgânicos, nomeadamente ρ-NΞCC6H4CCSiMe3, ρ-NΞCC6H4CCH e ρ-HCΞCC6H4CCH. No capítulo 3 descreve-se a síntese de diversos complexos monometálicos de Ferro (II) e ruténio (II) com os ligandos acima referidos e diferentes fragmentos metálicos [FeCp(CO)2]+, [FeCp(dppe)]+, [RuCp(dppe)]+ e [RuCp(PPh3)2]+. Os complexos obtidos foram caracterizados utilizando as técnicas espectroscópicas usuais: RMN, IV, UV-vis. A eficiência da interacção electrónica entre os centros, ao longo do sistema тт, foi avaliada por estudos electroquímicos (voltametria cíclica). No capítulo 4 encontram-se descritas as diversas tentativas de síntese dos compostos homo e heterobimetálicos. Os ligandos e fragmentos utilizados foram os mesmos que no capítulo anterior. O capítulo 5 descreve os pormenores experimentais da síntese e a caracterização espectroscópica dos compostos apresentados nos capítulos anteriores, bem como as condições gerais em que foram obtidos.

KrP laser CVD of chromium oxide by photodissociation of Cr(CO)(6)

This work reports on the synthesis of chromium oxide thin films prepared by photodissociation of Cr(CO)(6) in an oxidizing atmosphere, using a pulsed UV laser (KrF, lambda = 248 nm). The experimental conditions, which should enable the synthesis of CrO2, are discussed and results on the deposition of CrxOy films on Al2O3 (0001) substrates are presented.

Flavored asymmetries for type II seesaw leptogenesis

A novel contribution to the leptonic CP asymmetries in type II seesaw leptogenesis scenarios is obtained for the cases in which flavor effects are relevant for the dynamics of leptogenesis. In the so-called flavored leptogenesis regime, the interference between the tree-level amplitude of the scalar triplet decaying into two leptons and the one-loop wave function correction with leptons in the loop, leads to a new nonvanishing CP asymmetry contribution. The latter conserves total lepton number but violates lepton flavor. Cases in which this novel contribution may be dominant in the generation of the baryon asymmetry are briefly discussed.

Halogen-bonded tris (2,4-Bis(trichloromethyl)-1,3,5,triazapentadienato)-M(III) [M = Mn, Fe, Co] complexes and their catalytic activity in the peroxidative oxidation of 1-phenylethanol to acetophenone

One-pot template condensation of CCl3C=N with ammonia on a metal source [MnCl2 center dot 4H(2)O, FeCl3 center dot 6H(2)O or Co(CH3COO)(2)center dot 4H(2)O] in DMSO led to the formation of tris(2,4-bis(trichloromethyl)-1,3,5-triazapentadienato)-M(III) complexes, [M(NH=C(CCl3)NC(CCl3)=-NH}(3)]center dot n(CH3)(2)SO [M = Mn, n = 1 (1); M = Fe, n = 2 (2); M = Co, n = 2 (3)1, which were characterized using elemental analysis, and IR, ESI-MS and single-crystal X-ray analysis. The role of inter- and intramolecular non-covalent halogen and hydrogen bonds in the synthesis of 1-3 is discussed. It is shown that the crystal ionic radii of the metal ions [68.5 (Co) < 69 (Fe) < 72 (Mn), pm] are related to the corresponding Cl center dot center dot center dot Cl distances [3.178 (3) > 3.155 (2) > 3.133 (1) Al. Compounds 1-3 and the related di(triazapentadienato)-Cu(v) complex [Cu(NH=C(CCl3)NC(CCl3)=NH}2]center dot 2(CH3)(2)SO (4) act as catalyst precursors for the additive-free microwave (MW) assisted homogeneous oxidation of 1-phenylethanol with tert-butylhydroperoxide (TBHP), leading to the formation of acetophenone with yields up to 99% and TONs up to 5.0 x 10(3) after 1 h of low power (10 W) MW irradiation.

Mu-Chlorido-Bridged Dimanganese(II) complexes of the schiff base derived from [2+2] condensation of 2,6-diformyl-4-methylphenol and 1,3-bis(3-aminopropyl)tetramethyldisloxane: structure, magnetismo, electrochemical behaviour, and catalytic oxidation of secondary alcohols

The reaction of 2,6-diformyl-4-methylphenol with 1,3-bis(3-aminopropyl)tetramethyldisiloxane in the presence of MnCl2 in a 1:1:2 molar ratio in methanol afforded a dinuclear -chlorido-bridged manganese(II) complex of the macrocyclic [2+2] condensation product (H2L), namely, [Mn2Cl2(H2L)(HL)]Cl center dot 3H(2)O (1). The latter afforded a new compound, namely, [Mn2Cl2(H2L)(2)][MnCl4]center dot 4CH(3)CN center dot 0.5CHCl(3 center dot)0.4H(2)O (2), after recrystallisation from 1:1 CHCl3/CH3CN. The co-existence of the free and complexed azomethine groups, phenolato donors, mu-chlorido bridges, and the disiloxane unit were well evidenced by ESI mass spectrometry and FTIR spectroscopy and confirmed by X-ray crystallography. The magnetic measurements revealed an antiferromagnetic interaction between the two high-spin (S = 5/2, g = 2) manganese(II) ions through the mu-chlorido bridging ligands. The electrochemical behaviour of 1 and 2 has been studied, and details of their redox properties are reported. Both compounds act as catalysts or catalyst precursors in the solvent-free low-power microwave-assisted oxidation of selected secondary alcohols, for example, 1-phenylethanol, cyclohexanol, 2- and 3-octanol, to the corresponding ketones in the absence of solvent. The highest yield of 72% was achieved for 1-phenylethanol by using a maximum of 1% molar ratio of catalyst relative to substrate.

Modelação estatística das co-infecções da SIDA em países da União Europeia

Dissertação apresentada para obtenção do Grau de Doutor em Matemática, especialidade de Estatística, pela Universidade Nova de Lisboa, Faculdade de Ciências e Tecnologia

Tetranuclear COPPER(II) complexes with macrocyclic and open-chain disiloxane ligands as catalyst precursors for hydrocarboxylation and oxidation of alkanes and 1-phenylethanol

Two new tetranuclear complexes [Cu-4(mu-O)(L-1)-Cl-4] and [Cu-4(mu(4)-O)(L-2)(2)Cl-4] (2), where H2L1 is a macrocyclic ligand resulting from [2+2] condensation of 2,6-diformy1-4-methylphanol (DFF) and 1,3-bis(aminopropy1)tetramethyldisiloxane, and HL2 is a 1:2 condensation product: of DFF with trimethylsilyl p-aminobenzoate, have been prepared. The structures of the products were established by Xray diffraction. The complexes have been characterised by FTIR, UV/Vis spectroscopy, ES1 mass-spectrometry and magnetic susceptibility measurements. The latter revealed that the letrftriuclear complexes can be descr bed as two ferromagnetically coupled dinuclear units, in which the two copper(II) ions interact antiferromacinetically. The ccimpi.iunds act as homogeneous catalyst precursors for a number of single-pot reactions, including (I) hydrocarbaxylation, with CO, H2O and K2S2O8, of a variety of linear and cyclic (n = 5-8) alkanes into the corresponding Cn+1 carboxylic acids, (ii) peroxidative oxidation of cyclohexane, and (iii) solvent-free microwave-assisted oxidation of 1-phenyletha.nol.