Evaluation of estrogen neuroprotective effect on nigrostriatal dopaminergic neurons following 6-hydroxydopamine injection into the substantia nigra pars compacta or the medial forebrain bundle

Studies involving estrogen treatment of ovariectomized rats or mice have attributed to this hormone a neuroprotective effect on the substantia nigra pars compacta (SNpc) neurons. We investigated the effect of estradiol replacement in ovariectomized rats on the survival of dopaminergic mesencephalic cell and the integrity of their projections to the striatum after microinjections of 1 mu g of 6-hydroxydopamine (6-OHDA) into the right SNpc or medial forebrain bundle (MFB). Estradiol replacement did not prevent the reduction either in the striatal concentrations of DA and metabolites or in the number of nigrostriatal dopaminergic neurons following lesion with 1 mu g of 6-OHDA into the SNpc. Nevertheless, estradiol treatment reduced the decrease in striatal DA following injection of 1 mu g of 6-OHDA into the MFB. Results suggest therefore that estrogen protect nigrostriatal dopaminergic neurons against a 6-OHDA injury to the MFB but not the SNpc. This may be due to the distinct degree of lesions promoted in these different rat models of Parkinson`s disease.

A novel method for solid-phase synthesis of oligosaccharides using the N-1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde) linker

The application of the N-1-(4,4-dimethyl-2,6-dioxocyclohexylidene)ethyl (Dde) linker for the solid-phase synthesis of oligosaccharides is described. The oligosaccharide products can be cleaved from the resin by hydrazine, ammonia or primary amines, but the linker is stable under the conditions of oligosaccharide synthesis. The first sugar can be attached to the resin linker via a vinylogous amide bond, or by ether linkage using a p-aminobenzyl alcohol converter. (C) 2001 Elsevier Science Ltd. All rights reserved.

Outcomes of transient evoked otoacoustic emission testing in 6-year-old school children: A comparison with pure tone screening and tympanometry

Objectives: (1) To establish test performance measures for Transient Evoked Otoacoustic Emission testing of 6-year-old children in a school setting; (2) To investigate whether Transient Evoked Otoacoustic Emission testing provides a more accurate and effective alternative to a pure tone screening plus tympanometry protocol. Methods: Pure tone screening, tympanometry and transient evoked otoacoustic emission data were collected from 940 subjects (1880 ears), with a mean age of 6.2 years. Subjects were tested in non-sound-treated rooms within 22 schools. Receiver operating characteristics curves along with specificity, sensitivity, accuracy and efficiency values were determined for a variety of transient evoked otoacoustic emission/pure tone screening/tympanometry comparisons. Results: The Transient Evoked Otoacoustic Emission failure rate for the group was 20.3%. The failure rate for pure tone screening was found to be 8.9%, whilst 18.6% of subjects failed a protocol consisting of combined pure tone screening and tympanometry results. In essence, findings from the comparison of overall Transient Evoked Otoacoustic Emission pass/fail with overall pure tone screening pass/fail suggested that use of a modified Rhode Island Hearing Assessment Project criterion would result in a very high probability that a child with a pass result has normal hearing (true negative). However, the hit rate was only moderate. Selection of a signal-to-noise ratio (SNR) criterion set at greater than or equal to 1 dB appeared to provide the best test performance measures for the range of SNR values investigated. Test performance measures generally declined when tympanometry results were included, with the exception of lower false alarm rates and higher positive predictive values. The exclusion of low frequency data from the Transient Evoked Otoacoustic Emission SNR versus pure tone screening analysis resulted in improved performance measures. Conclusions: The present study poses several implications for the clinical implementation of Transient Evoked Otoacoustic Emission screening for entry level school children. Transient Evoked Otoacoustic Emission pass/fail criteria will require revision. The findings of the current investigation offer support to the possible replacement of pure tone screening with Transient Evoked Otoacoustic Emission testing for 6-year-old children. However, they do not suggest the replacement of the pure tone screening plus tympanometry battery. (C) 2001 Elsevier Science Ireland Ltd. All rights reserved.

Syntaxin 7 complexes with mouse Vps10p tail interactor 1b, Syntaxin 6, vesicle-associated membrane protein (VAMP)8, and VAMP7 in B16 melanoma cells

Syntaxin 7 is a mammalian target soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) involved in membrane transport between late endosomes and lysosomes. The aim of the present study was to use immunoaffinity techniques to identify proteins that interact with Syntaxin 7. We reasoned that this would be facilitated by the use of cells producing high levels of Syntaxin 7, Screening of a large number of tissues and cell lines revealed that Syntaxin 7 is expressed at very high levels in B16 melanoma cells. Moreover, the expression of Syntaxin 7 increased in these cells as they underwent melanogenesis. From a large scale Syntaxin 7 immunoprecipitation, we have identified six polypeptides using a combination of electrospray mass spectrometry and immunoblotting. These polypeptides corresponded to Syntaxin 7, Syntaxin 6, mouse Vps10p tail interactor 1b (mVti1b), alpha -synaptosome-associated protein (SNAP), vesicle-associated membrane protein (VAMP)8, VAMP7, and the protein phosphatase 1M regulatory subunit. We also observed partial colocalization between Syntaxin 6 and Syntaxin 7, between Syntaxin 6 and mVti1b, but not between Syntaxin 6 and the early endosomal t-SNARE Syntaxin 13. Based on these and data reported previously, we propose that Syntaxin 7/mVti1b/Syntaxin 6 may form discrete SNARE complexes with either VAMP7 or VAMPS to regulate fusion events within the late endosomal pathway and that these events may play a critical role in melanogenesis.

2-pyrazinylnitrene and 4-pyrimidylnitrene. Ring expansion to 1,3,5-triazacyclohepta-1,2,4,6-tetraene and ring opening to (2-isocyanovinyl)carbodiimide

Tetrazolo[1,5-a]pyrazine/2-azidopyrazine 9T/9A undergo photolysis in Ar matrix at cryogenic temperatures to yield 1,3,5-triazacyclohepta-1,2,4,6-tetraene 21 as the first observable intermediate, and 1-cyanoimidazole 11 and (2-isocyanovinyl)carbodiimide 22 as the final products. The latter tautomerizes to 2-(isocyanovinyl)cyanamide 23 on warming to 40 K. The same intermediate 21 and the same final products are obtained on matrix photolysis of the isomeric tetrazolo[1,5-c]pyrimidine/4-azidopyrimidine 24T/24A. These photolysis results as well as those of the previously reported thermal ring contraction of N-15-labeled 2-pyrazinyl- and 4-pyrimidylnitrenes to 1-cyanoimidazoles can all be rationalized in terms of selective ring opening of 21 or nitrine 10 to a nitrile ylide zwitterion 28 prior to formation of the final products, 11 and 22. The results are supported by high-level ab initio and DFT calculations (CASPT2-CASSCF(6,6), G3(MP2), and B3LYP/6-31+G*) of the energies and IR spectra of the intermediates and products.

Production of the baculovirus-expressed dengue virus glycoprotein NS1 can be improved dramatically with optimised regimes for fed-batch cultures and the addition of the insect moulting hormone, 20-Hydroxyecdysone

A perennial problem in recombinant protein expression is low yield of the product of interest. A strategy which has been shown to increase the production of baculovirus-expressed proteins is to utilise fed-batch cultures. One disadvantage of this approach is the time-consuming task of optimising the feeding strategy. Previously, a statistical optimisation routine was applied to develop a feeding strategy that increased the yield of beta-Galactosidase (beta-Gal) by 2.4-fold (Biotechnol. Bioeng, 59 (1998) 178). This involves the single addition of nutrient concentrates (amino acids, lipids. glucose and yeastolate ultrafiltrate) into Sf9 cell cultures grown in SF900II medium. In this study, it is demonstrated that this optimised fed-batch strategy developed for a high-yielding intracellular product beta-Gal could be applied successfully to a relatively low-yielding glycosylated and secreted product such as the dengue virus glycoprotein NS1. Optimised batch infections yielded 4 mug/ml of NS1 at a peak cell density of 4.2 x 10(6) cells/ml. In contrast. optimised fed-batch infections exhibited a 3-fold improvement in yield, with 12 mug ml of NS1 produced at a peak cell density of 11.3 x 10(6) cells/ml. No further improvements in yield were recorded when the feed volumes were doubled and the peak cell density was increased to 23 x 10(6) cells/ml, unless the cultures were stimulated by the addition of 4 mug/ml of 20-Hydroxyecdysone (an insect moulting hormone). In this case, the NS1 yield was increased to 20 mug/ml. which was nearly 5-fold higher than optimised batch cultures. (C) 2002 Elsevier Science B.V. All rights reserved.

Quantitative trait loci for agronomic and seed quality traits in an F-2 and F-4 : 6 soybean population

Molecular breeding is becoming more practical as better technology emerges. The use of molecular markers in plant breeding for indirect selection of important traits can favorably impact breeding efficiency. The purpose of this research is to identify quantitative trait loci (QTL) on molecular linkage groups (MLG) which are associated with seed protein concentration, seed oil concentration, seed size, plant height, lodging, and maturity, in a population from a cross between the soybean cultivars 'Essex' and 'Williams.' DNA was extracted from F-2 generation soybean leaves and amplified via polymerase chain reaction (PCR) using simple sequence repeat (SSR) markers. Markers that were polymorphic between the parents were analyzed against phenotypic trait data from the F-2 and F-4:6 generation. For the F-2 population, significant additive QTL were Satt540 (MLG M, maturity, r(2)=0.11; height, r(2)=0.04, seed size, r(2)=0.061, Satt373 (MLG L, seed size, r(2)=0.04; height, r(2)=0.14), Satt50 (MLG A1, maturity r(2)=0.07), Satt14 (MLG D2, oil, r(2)=0.05), and Satt251 (protein r(2)=0.03, oil, r(2)=0.04). Significant dominant QTL for the F-2 population were Satt540 (MLG M, height, r(2)=0.04; seed size, r(2)=0.06) and Satt14 (MLG D2, oil, r(2)=0.05). In the F-4:6 generation significant additive QTL were Satt239 (MLG I, height, r(2)=0.02 at Knoxville, TN and r(2)=0.03 at Springfield, TN), Satt14 (MLG D2, seed size, r(2)=0.14 at Knoxville, TN), Satt373 (MLG L, protein, r(2)=0.04 at Knoxville, TN) and Satt251 (MLG B I, lodging r(2)=0.04 at Springfield, TN). Averaged over both environments in the F-4:6 generation, significant additive QTL were identified as Satt251 (MLG B 1, protein, r(2)=0.03), and Satt239 (MLG 1, height, r(2)=0.03). The results found in this study indicate that selections based solely on these QTL would produce limited gains (based on low r(2) values). Few QTL were detected to be stable across environments. Further research to identify stable QTL over environments is needed to make marker-assisted approaches more widely adopted by soybean breeders.

Description of stable pain in rheumatoid arthritis: A 6 year study

Objective. To study pain quality and variability in patients with rheumatoid arthritis (RA). Methods. Pain, disease activity, and functional status Were assessed 3 times over 6 years in an initial cohort of 120 clinic patients with chronic pain from RA. A pain visual analog scale and the McGill Pain Questionnaire (MPQ) were used to record pain intensity and quality. RA disease activity and function were measured. Results. There was no statistically significant difference in any measure over the 3 assessments. RA pain intensity was moderate. The MPQ showed that sensory components of the pain were described in terms of pressure and constriction. Pain related affect was described with adjectives suggesting positive psychological adaptation to pain. Conclusion. The. results indicate a general profile of no change in pain sensation, affect, and emotional quality in clinic monitored patients with ongoing RA and ongoing, moderate levels of disease activity and function. The MPQ provides qualitative detail to patient's report of pain severity that could be a useful addition to longterm documentation of RA outcome. Regular MPQ documentation of current pain in outpatients could indicate whether any significant change in pain levels is reflected in altered word selection that reflects physiological or psychological change, and could assist clinicians to select the most appropriate form of therapy for RA pain.

4,6,8,10,16-penta- and 4,6,8,10,16,18-hexamethyldocosanes from the cane beetle Antitrogus parvulus - Cuticular hydrocarbons with unprecedented structure and stereochemistry

[GRAPHICS] The major cuticular hydrocarbons from the cane beetle species Antitrogus parvulus were deduced to be 4,6,8,10,16,18-hexa- and 4,6,8,10,16-pentamethyldocosanes 2 and 3, respectively. Isomers of 2,4,6,8-tetramethylundecanal 27, 36, and 37, derived from 2,4,6-trimethylphenol, were coupled with the phosphoranes 28 and 29 to furnish alkenes and, by reduction, diastereomers of 2 and 3. Chromatographic and spectroscopic comparisons confirmed 2 as either 6a or 6b and 3 as either 34a or 34b.

Optimising high-intensity treadmill training using the running speed at maximal O-2 uptake and the time for which this can be maintained

The aim of this study was to compare the effects of two high-intensity, treadmill interval-training programs on 3000-m and 5000-m running performance. Maximal oxygen uptake ((V) over dot O-2max), the running speed associated with (V) over dot O-2max (nu (V) over dot O-2max), the time for which nu (V) over dot O-2max can be maintained (T-max), running economy (RE), ventilatory threshold (VT) and 3000-m and 5000-m running times were determined in 27 well-trained runners. Subjects were then randomly assigned to three groups; (1) 60% T-max (2) 70% T-max and (3) control. Subjects in the control group continued their normal training and subjects in the two T-max groups undertook a 4-week treadmill interval-training program with the intensity set at nu (V) over dot O-2max and the interval duration at the assigned T-max. These subjects completed two interval-training sessions per week (60% T-max = six intervals/session, 70% T-max group = five intervals/session). Subjects were re-tested on all parameters at the completion of the training program. There was a significant improvement between pre- and post-training values in 3000-m time trial (TT) performance in the 60% T-max group compared to the 70% T,,a, and control groups [mean (SE); 60% T-max = 17.6 (3.5) s, 70% T-max = 6.3 (4.2) s, control = 0.5 (7.7) s]. There was no significant effect of the training program on 5000-m TT performance [60% T-max = 25.8 (13.8) s, 70% T-max = 3.7 (11.6) s, control = 9.9 (13.1) s]. Although there were no significant improvements in (V) over dot O-2max, nu (V) over dot (2max) and RE between groups, changes in (V) over dot O-2max and RE were significantly correlated with the improvement in the 3000-m TT. Furthermore, VT and T-max were significantly higher in the 60% Tmax group post-compared to pre-training. In conclusion, 3000-m running performance can be significantly improved in a group of well-trained runners, using a 4-week treadmill interval training program at nu (V) over dot O-2max with interval durations of 60% T-max.

The preparation of zinc(II) and cadmium(II) complexes of the pentadentate N3S2 ligand formed from 2,6-diacetylpyridine and S-benzyldithiocarbazate (H2SNNNS) and the X-ray crystal structure of the novel dimeric [Zn2(SNNNS)2] complex

The pentadentate chelating agent, 2,6-diacetylpyridinebis(S-benzyldithiocarbazate) (H2SNNNS) reacts with zinc(II) and cadmium(II) ions forming stable complexes of empirical formula, [M(SNNNS)] (M=Zn2+, Cd2+; SNNNS2 =doubly deprotonated anionic form of the Schiff base). These complexes have been characterized by a variety of physico-chemical techniques. IR and H-1 NMR spectral evidence indicate that the Schiff base coordinates to the zinc(II) and cadmium(II) ions via the pyridine nitrogen atoms, the azomethine nitrogen atoms and the mercaptide sulfur atoms. The crystal and molecular structure of the zinc(II) complex has been determined by X-ray diffraction. The complex is a dimer in which the pyridine nitrogen atom,the azomethine nitrogen atom and the thiolate sulfur atom from one ligand coordinate to one of the zinc(II) ions whereas the azomethine and thiolate sulfur atoms from another ligand complete pentacoordination around the zinc(II) ion, the ligands being coordinated in their deprotonated forms. The coordination geometry about each zinc(II) can be considered as intermediate between a square-pyramid and trigonal-bipyramid. The cadmium(II) complex is also assigned with a dimeric structure. (C) 2003 Elsevier Ltd. All rights reserved.

Estratégias de cálculo mental de alunos da 5ª série/6º ano do ensino fundamental

Esta pesquisa de mestrado teve como principal objetivo investigar estratégias de cálculo mental, utilizadas por alunos de uma 5ª série/6º ano do ensino fundamental ao resolver cálculos de adição e subtração. Para atingir este objetivo procuramos responder aos questionamentos: Quais estratégias de cálculo mental, alunos da 5ª série/6º ano empregam na resolução de cálculos de adição e subtração? Que relações existem entre o tipo de cálculo envolvido e a estratégia adotada para resolvê-lo? Para respondermos a essas questões, seguimos uma metodologia de natureza qualitativa, configurada como estudo de caso do tipo etnográfico. O trabalho de campo foi desenvolvido em uma turma de 5ª série/6º ano do ensino fundamental de uma escola pública da rede estadual de ensino do município de Serra. A pesquisa aconteceu de maio a dezembro de 2013. Oito alunos resolveram uma atividade diagnóstica composta de quatro sequências de cálculos mentais, a saber, fatos fundamentais do número 5, do número 10, do número 20 e do número 100, dentre adições e subtrações próximas a esses resultados. Todos alunos participaram da etapa de entrevistas. Dos oito alunos, foram escolhidos dados de três que participaram de outras etapas da pesquisa. Os registros realizados pelos alunos na etapa de observação da turma, na etapa diagnóstica e na etapa de intervenção didática, as anotações no caderno de campo e algumas gravações em áudio serviram como fontes de coleta de dados. Utilizamos as estratégias identificadas por Beishuizen (1997), Klein e Beishuizen (1998), Thompson (1999, 2000) e Lucangeli et al. (2003), como categorias de análise. Através da análise de dados, constatamos que as escolhas das estratégias de cálculo mental pelos alunos variaram de acordo com o tipo de sequência de cálculos, a operação aritmética (adição ou subtração) e o estado emocional deles durante a atividade. Foi possível identificar o uso de duas estratégias combinadas, o algoritmo mental e estratégias de contagens nos dedos para grande parte dos cálculos. O uso do algoritmo mental mostrou-se um procedimento de grande sobrecarga mental e, em alguns cálculos de adição sem reserva, serviu apenas como apoio à visualização numérica, sendo executado pelo aluno da esquerda para a direita, semelhantemente à estratégia de decomposição numérica. Os dados deste estudo apontam para: (i) a necessidade de se trabalhar fatos numéricos fundamentais de adição e subtração via cálculo mental de maneira sistemática em sala de aula; (ii) a necessidade de se ensinar estratégias autênticas de cálculo mental para que os alunos não se tornem dependentes de estratégias como contagens e algoritmo mental, que são mais difíceis de serem executadas com êxito; (iii) a importância de entrevistar, individualmente, os alunos a fim de compreender e avaliar o desenvolvimento destes em tarefas de cálculo mental.

Influência de polimorfismos nos genes FcγRIIa, CD209, VDR, TNF-α, IL-4, IL-6 e INF-γ na persistência de sintomas clínicos da dengue na fase de covalescença

Diferenças na susceptibilidade do hospedeiro à infecção, na gravidade e na permanência do quadro clínico da doença podem ser atribuídas, em parte, às variações da resposta imune. Estas variações são associadas a polimorfismos de nucleotídeo único (do inglês: single nucleotide polymorphisms - SNPs). Como estudo prévio, foi realizada a caracterização da população geral do Espírito Santo (ES) - Brasil e de uma subpopulação do estado, de origem Pomerana, quanto aos SNPs -131 H/R, -336 A/G, TaqI, -308 A/G, -590 C/T, -174 G/C e +874 A/T nos genes FcγRIIa, CD209, VDR, TNFα, IL-4, IL-6 e INF-γ, respectivamente. Cem indivíduos da Grande Vitória representaram a população geral do ES e 59 indivíduos de Santa Maria de Jetibá representaram a população de origem Pomerana. Como a fase aguda da dengue é bem caracterizada, este estudo objetivou ampliar o conhecimento da fase de convalescença. Noventa e seis indivíduos diagnosticados com dengue sintomática no final de 2012 e início de 2013, no ES, foram acompanhados por 60 dias a partir do início dos sintomas por meio do preenchimento de um questionário clínico e epidemiológico em quatro entrevistas. A persistência de 37 sintomas clínicos da dengue foi avaliada. Para analisar a influência da genética do sistema imunológico do hospedeiro na persistência de sintomas clínicos da dengue na fase de convalescença, foi determinada a associação entre os sete SNPs, para os quais a população do ES foi caracterizada, e a persistência de sintomas. O DNA genômico dos participantes do estudo foi extraído do sangue periférico e a genotipagem dos SNPs foi realizada por reação em cadeia da polimerase - polimorfismo de comprimento de fragmento de restrição (do inglês: polymerase chain reaction - restriction fragment length polymorphism - PCR-RFLP) As frequências genotípicas de todos os SNPs encontraram-se em equilíbrio de Hardy-Weinberg (do inglês: Hardy-Weinberg equilibrium - HWE), com exceção do SNP no gene IL-6. Não houve diferença estatisticamente significante nas frequências genotípicas dos SNPs nos genes FcγRIIa, CD209, VDR, TNF-α e IL-4 entre as duas populações. Diferença estatisticamente significante foi encontrada entre as duas populações nas distribuições genotípicas dos SNPs nos genes IL-6 (p = 0,03) e INF-γ (p = 0,007). Trinta e sessenta dias após o início dos sintomas, 38,5% e 11,5% dos indivíduos com dengue sintomática reportaram ter pelo menos um sintoma clínico da dengue, respectivamente. Dos sintomas analisados, os mais persistentes foram os relacionados à síndrome da fadiga como mialgia, artralgia, astenia e mal-estar, sendo a mialgia o mais frequente. A persistência de sintomas em 30 dias foi associada ao gênero feminino (p = 0,044) e a persistência de sintomas constitucionais foi associada à dengue secundária (p = 0,041). O SNP no gene FcγRIIa, foi associado à persistência de sintomas em 30 dias, no subgrupo de indivíduos com dengue secundária (p = 0,046), sendo a presença do alelo H associada à não persistência de sintomas (p = 0,014). A presença do alelo A do SNP no gene TNF-α foi associada à não persistência de sintomas no subgrupo de indivíduos com dengue secundária (p = 0,025), sendo o genótipo GG associado à persistência de sintomas neurológicos, psicológicos e comportamentais em 30 dias (p = 0,038). A presença do alelo C do SNP no gene IL-6 foi associado à persistência de sintomas dermatológicos em 30 dias (p = 0,005). O perfil genético desses SNPs pode favorecer o estabelecimento de marcadores imunogenéticos associados à fase convalescente da infecção pelo vírus da dengue (do inglês: dengue virus - DENV).

Show me your body and I tell you how hold you are: a non-invasive methodto define 6 life history-classes in Risso's dolphins (Grampus griseus), using an identified trial population in the Atlantic

European Cetacean Society Conference Workshop, Galway, Ireland, 25th March 2012.

Synthesis, characterization, electrochemical behavior and in vitro protein tyrosine kinase inhibitory activity of the cymene-halogenobenzohydroxamato [Ru(eta(6)-cymene)(bha)Cl] complexes

The ruthenium(II)-cymene complexes [Ru(eta(6)-cymene)(bha)Cl] with substituted halogenobenzohydroxamato (bha) ligands (substituents = 4-F, 4-Cl, 4-Br, 2,4-F-2, 3,4-F-2, 2,5-F-2, 2,6-F-2) have been synthesized and characterized by elemental analysis, IR, H-1 NMR, C-13 NMR, cyclic voltammetry and controlled-potential electrolysis, and density functional theory (DFT) studies. The compositions of their frontier molecular orbitals (MOs) were established by DFT calculations, and the oxidation and reduction potentials are shown to follow the orders of the estimated vertical ionization potential and electron affinity, respectively. The electrochemical E-L Lever parameter is estimated for the first time for the various bha ligands, which can thus be ordered according to their electron-donor character. All complexes exhibit very strong protein tyrosine kinase (PTK) inhibitory activity, even much higher than that of genistein, the clinically used PTK inhibitory drug. The complex containing the 2,4-difluorobenzohydroxamato ligand is the most active one, and the dependences of the PTK activity of the complexes and of their redox potentials on the ring substituents are discussed. (C) 2012 Elsevier B.V. All rights reserved.