An oriental melon 9-lipoxygenase gene CmLOX09 response to stresses, hormones, and signal substances / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

In plants, lipoxygenases (LOXs) play a crucial role in biotic and abiotic stresses. In our previous study, five 13-LOX genes of oriental melon were regulated by abiotic stress but it is unclear whether the 9-LOX is involved in biotic and abiotic stresses. The promoter analysis revealed that CmLOX09 (type of 9-LOX) has hormone elements, signal substances, and stress elements. We analyzed the expression of CmLOX09 and its downstream genes-CmHPL and CmAOS-in the leaves of four-leaf stage seedlings of the oriental melon cultivar "Yumeiren" under wound, hormone, and signal substances. CmLOX09, CmHPL, and CmAOS were all induced by wounding. CmLOX09 was induced by auxin (indole acetic acid, IAA) and gibberellins (GA3); however, CmHPL and CmAOS showed differential responses to IAA and GA3. CmLOX09, CmHPL, and CmAOS were all induced by hydrogen peroxide (H2O2) and methyl jasmonate (MeJA), while being inhibited by abscisic acid (ABA) and salicylic acid (SA). CmLOX09, CmHPL, and CmAOS were all induced by the powdery mildew pathogen Podosphaera xanthii. The content of 2-hexynol and 2-hexenal in leaves after MeJA treatment was significantly higher than that in the control. After infection with P. xanthii, the diseased leaves of the oriental melon were divided into four levels-levels 1, 2, 3, and 4. The content of jasmonic acid (JA) in the leaves of levels 1 and 3 was significantly higher than that in the level 0 leaves. In summary, the results suggested that CmLOX09 might play a positive role in the response to MeJA through the hydroperoxide lyase (HPL) pathway to produce C6 alcohols and aldehydes, and in the response to P. xanthii through the allene oxide synthase (AOS) pathway to form JA.

Suicide attempts among children and adolescents: partial or total injury? / Tentativa de suicídio infanto-juvenil: lesão da parte ou do todo?

This study sought to verify the records on file and the number of cases of attempted suicide among children and adolescents who were attended by Emergency Care health professionals in the municipality of Matozinhos, Minas Gerais, Brazil. Documentary and descriptive research was conducted, the data for which was collected by means of an investigation of Outpatient Records from 2008 to 2010. Of the 73,000 files evaluated, those dealing with cases of attempted suicide among children and adolescents between the age of 3 and 18 years were selected. It was revealed that the health professionals, particularly physicians and nurses, fail to register the cases appropriately, invalidating information about the problem and potential prevention measures. The conclusion reached was that underreporting and the discrepancy of the diagnoses which were not duly referred to the competent agencies require rethinking and reviewing medical practices, and taking a systematic and careful look to address the individual as a complex whole.

Neste estudo procurou-se verificar o registro e o número de casos de tentativa de suicídio entre crianças e adolescentes do município de Matozinhos, Minas Gerais, Brasil, que foram atendidos pelos profissionais de saúde do Pronto-Atendimento. Trata-se de uma pesquisa documental e descritiva, cuja coleta dos dados ocorreu por meio de investigação nas Fichas Ambulatoriais, no período de 2008 a 2010. Das 73.000 fichas levantadas, selecionaram-se aquelas que tratavam de casos de tentativa de suicídio entre crianças e adolescentes do município, com idades entre três e 18 anos. Percebeu-se que os profissionais de saúde, mais especificamente os médicos e enfermeiros, não registram os casos de forma adequada, inviabilizando a informação sobre o problema e as medidas de prevenção. Concluiu-se que a subnotificação, a discrepância dos diagnósticos e o não encaminhamento aos órgãos competentes exigem repensar e rever a prática médica e dirigir um olhar sistematizado e cuidadoso para perceber o sujeito como um todo complexo.

Serological detection of Toxoplasma gondii, Leishmania infantum and Neospora caninum in cats from an area endemic for leishmaniasis in Brazil / Detecção sorológica de Toxoplasma gondii, Leishmania infantum e Neospora caninum em gatos de uma área endêmica para leishmaniose no Brasil

An investigation was made into the occurrence of antibodies to Toxoplasma gondii, Leishmania infantum and Neospora caninum in 151 domestic cats, based on the indirect fluorescent antibody test (IFAT). Serum samples were collected from 151 domestic cats (65 free-roaming and 86 domiciled cats; 55 males and 96 females) in Campo Grande, Mato Grosso do Sul, Brazil between January and April 2013. IgG antibodies to T. gondii, L. infantum and N. caninum were found, respectively, in 49 (32.5%), 34 (22.5%) and 10 (6.6%) sampled cats. A positive correlation was found between T. gondii and N. caninum, T. gondii and L. infantum, and N. caninum and L. infantum (p <0.05) infections. Also, a significant interaction was identified between gender and area of activity on the probability of T. gondii (p = 0.0324) infection. However, no significant interaction was observed between gender and area of activity on infections by either N. caninum or L. infantum. This study showed that cats from an area endemic for visceral leishmaniasis in Brazil are exposed to three different protozoans, two of which are causal agents of important zoonosis.

O presente estudo teve como objetivo investigar a ocorrência de anticorpos anti-Toxoplasma gondii, Leishmania infantum e Neospora caninum, em 151 gatos, por meio da Reação de Imunofluorescência Indireta (RIFI). Entre os meses de janeiro e abril de 2013, amostras de soro foram coletadas de 151 gatos domésticos (65 gatos errantes e 86 gatos domiciliados; 55 machos e 96 fêmeas), de Campo Grande, Mato Grosso do Sul, Brasil. Anticorpos IgG anti-T. gondii, anti-L. infantum e anti-N. caninum foram encontrados em 49 (32,5%), 34 (22,5%) e 10 (6,6%) gatos amostrados, respectivamente. Verificou-se uma associação estatisticamente significativa entre as infecções por T. gondii e N. caninum, T. gondii e L. infantum e N. caninum e Leishmania infantum (p <0,05). Além disso, foi observada uma interação significativa entre sexo, área de atividade na probabilidade de infecção por T. gondii (p = 0,0324). No entanto, não foi observada interação significativa entre sexo e área de atividade nas infecções por N. caninum e L. infantum. Este estudo mostrou que os gatos de uma área endêmica brasileira para leishmaniose visceral são expostos a três diferentes protozoários, sendo dois deles importantes agentes zoonóticos.

Modificação de proteínas por produtos de oxidação do colesterol: mecanismos e implicações biológicas / Modification of proteins by oxidation products of cholesterol: mechanisms and biological implications

O colesterol é um importante componente das membranas celulares em eucariotos superiores, desempenhando papéis estruturais e funcionais. O colesterol possui uma insaturação em sua estrutura sendo, portanto, alvo de oxidação mediada por espécies reativas de oxigênio e/ou nitrogênio. A oxidação não enzimática do colesterol gera, como produtos primários, os hidroperóxidos de colesterol. Tais moléculas, por sua vez, são altamente reativas e podem reagir com metais livres e/ou metaloproteínas, trazendo consequências à celula. Neste sentido, o primeiro capítulo deste trabalho tem como objetivo estudar a reação dos hidroperóxidos de colesterol (ChOOH) com o citocromo c (citc), uma heme proteína envolvida no transporte de elétrons na mitocôndria. Análises de espectroscopia no UV-Vis mostraram que o ChOOH promove o bleaching da banda Soret do citc de uma maneira dose-dependente. Mais ainda, esta reação leva à formação de radicais centrados em carbono tanto na proteína como no lipídeo, sugerindo uma redução homolítica do ChOOH. Como consequências, pode-se observar a oligomerização do citc, um processo que pode influenciar no transporte de elétrons bem como na sinalização para a apoptose. A partir da reação do citc com ChOOH podem surgir, direta ou indiretamente, outras espécies reativas, como aldeídos, cetonas e epóxidos. Dentre estas, destacam-se os aldeídos de colesterol, em particular o colesterol secoaldeído (CSec) e o carboxialdeído (ChAld), uma vez que foram encontrados elevados em placas ateroscleróticas e em tecidos cerebrais de pacientes com doenças neurodegenerativas. Tais espécies podem reagir com resíduos de aminoácidos provocando alterações estruturais e funcionais em proteínas. Neste sentido, o segundo capítulo deste trabalho tem como objetivo estudar a reação do ChAld com citc. Usando modelos mimétivos de membrana e espectrometria de massas, foi mostrado que o ChAld modifica covalentemente o citc por um mecanismo consistente com a formação de bases de Schiff. Tal modificação ocorre preferencialmente em resíduos de lisina que interagem com a membrana. Estas modificações influenciam na afinidade do citc pela membrana, aumentando sua aderência, o que pode ter influência no transporte de elétrons e sinalização para a apoptose. No terceiro e último capítulo deste trabalho nós buscamos uma ferramente analítica que permitisse analisar modificação de proteínas promovidas por produtos de oxidação de colesterol e outros esteróis. Em um estudo realizado em colaboração com o grupo do professor Porter na Universidade de Vanderbilt, utilizamos ensaios baseados em click chemistry para buscar proteínas modificadas. Para isso, foram sintetizados derivados de colesterol e 7-deidrocolesterol (7-DHC, precursor imediato do colesterol) contendo um grupo alquinil na sua cadeia lateral. Este grupo pode ser ligado a um grupo azida por meio de uma reação de cicloadição, em um processo conhecido como click chemistry. Após a síntese e caracterização dos derivados lipídicos contendo o grupo alquinil na cadeia lateral, células Neuro2a foram tratadas com o alquinil-7-DHC e o alquinil-colesterol para averiguar seu metabolismo. Análises por HPLC-MS/MS mostraram que ambos derivados contendo o grupo alquinil foram metabolisados e convertdos nos respectivos ésteres. Usando um modelo celular para a doença conhecida como Sindrome de Smith-Lemli-Opitz (SLOS), doença caracterizada pela deficiência na enzima 7-deidrocolesterol redutase, foi mostrado que o acúmulo característico de 7-DHC nos pacientes pode levar a uma maior modificação de proteínas promovidas por seus derivados, o que pode contribuir para o desenvolvimento da doença

Cholesterol is an important component of eukaryotic cellular membranes, where it has an influence in the fluidity and stability. Due to the presence of a double bond in its structure, cholesterol can be oxidized by reactive oxygen and nitrogen species. This non-enzymatic oxidation generates, as primary products, cholesterol hydroperoxides. Such molecules, in turn, are highly reactive and can react with free metal ions and/or metalloproteins, affecting cell metabolism. Therefore, the first chapter of the present study aims to investigate the reaction of cholesterol hydroperoxides (ChOOH) with cytochrome c (cytc), a heme protein involved in the mitochondrial electron transport. Spectroscopic analyses in the UV-Vis region showed that ChOOH induces a dose-dependent bleaching of cytc's Soret band. In addition, this reaction leads to the formation of carbon-centered radicals on both protein and lipid, suggesting a homolytic reduction of ChOOH. As consequences, cytc undergoes oligomerization, a process that can influence electron transport and apoptosis signaling. The reaction of cytc and ChOOH can produce, directly or indirectly, reactive species such as epoxides, aldehydes and ketones. Among them, cholesterol aldehydes, such as cholesterol secoaldehyde (CSec) and cholesterol carboxyaldehyde (ChAld), are of particular interest, since they were previously found elevated in atherosclerotic plaques and brain tissue of patients bearing neurodegenerative diseases. These species can also react with amino acid residues leading to protein denaturation and malfunction. With that in mind, the second chapter of this study aims to investigate the reaction of ChAld and cytc. Using mimetic membrane models and mass spectrometry analyses, we showed that ChAld covalently modifies cytc through a mechanism consistent with the formation of Schiff base adducts. Such modification occurs mostly at lysine residues that are known to interact with the membrane. The modifications have an influence in the affinity of cytc to the membrane, where they increase its binding to the membrane, a process that could affect the electron transport and apoptosis signaling. In the last and third chapter of this study we wanted an analytical tool that allowed the investigation of protein adduction promoted by cholesterol and other sterols-derived oxidation products. In a study performed in collaboration with the Porter group from Vanderbilt University, we used analyses based on click chemistry to search for protein adduction. To address that, we first synthesized derivatives of cholesterol and 7-dehydrocholesterol (7-DHC, the immediate precursor of cholesterol) containing an alkynyl group in the side chain. The alkynyl group can be ligated to an azide group through a cycloaddition reaction, in a process known as click chemistry. After the synthesis and characterization of alkynyl derivatives, Neuro2a cells were treated with alkynyl-7-DHC and alkynyl-cholesterol to check their metabolism. HPLC-MS/MS analyses showed that both alkynyl derivatives are metabolized and converted into their respective esters. In addition, using a cell model for Smith-Lemli-Optiz Syndrome (SLOS), a disease characterized by the deficiency in the dehydrocholesterol reductase 7, we showed that the characteristic accumulation of 7-DHC in SLOS patients might be associated with protein adduction promoted by its oxidation products, which might contribute to the development of the disease

Water sorption of CH3- and CF3-Bis-GMA based resins with additives

OBJECTIVES: To evaluate the effect of additives on the water sorption characteristics of Bis-GMA based copolymers and composites containing TEGDMA, CH3Bis-GMA or CF3Bis-GMA. MATERIAL AND METHODS: Fifteen experimental copolymers and corresponding composites were prepared combining Bis-GMA and TEGDMA, CH3Bis-GMA or CF3Bis-GMA, with aldehyde or diketone (24 and 32 mol%) totaling 30 groups. For composites, barium aluminosilicate glass and pyrogenic silica was added to comonomer mixtures. Photopolymerization was effected by 0.2 wt% each of camphorquinone and N,N-dimethyl-p-toluidine. Specimen densities in dry and water saturated conditions were obtained by Archimedes' method. Water sorption and desorption were evaluated in a desorption-sorption-desorption cycle. Water uptake (%WU), water desorption (%WD), equilibrium solubility (ES; µg/mm³), swelling (f) and volume increase (%V) were calculated using appropriate equations. RESULTS: All resins with additives had increased %WU and ES. TEGDMA-containing systems presented higher %WU, %WD, ES, f and %V values, followed by resins based on CH3Bis-GMA and CF3Bis-GMA. CONCLUSIONS: Aldehyde and diketone led to increases in the water sorption characteristics of experimental resins.

Preservation of Haemal Nodes Fixed by Means of Two Different Aldehyde Fixatives: a Scanning Electron Microscopic Study / Preservación de Nodos Linfáticos Hemales Fijados por Dos Fijadores de Aldehído Diferentes: un Estudio de Microscopía Electrónica de Barrido

Glutaraldehyde is the fixative most commonly used in electron microscope studies of biological tissues, however it is often necessary to use samples which were not fixed in this fixative, even with the usual uncertainty of the results that may be obtained. The fixation is the more delicate step of the sample processing. Therefore in this work, the quality of preservation of haemal nodes fixed with two classic aldehyde fixatives: formaldehyde and glutaraldehyde we have compared under the scanning electron microscope. Our results showed that both fixatives were successful in preserving the morphology of haemal nodes components; however glutaraldehyde conferred satisfactory results mainly in the preservation of parenchymal cells, whereas formaldehyde was better for preservation of stromal fibres.

El glutaraldehido es el fijador que se utiliza con más frecuencia en estudios en los tejidos biológicos a través microscopía electrónica. Sin embargo, a menudo es necesario utilizar muestras que no han sido fijadas con este fijador, aún con la incertidumbre de los resultados que se puedan obtener. La fijación es el paso más importante en el procesamiento de los tejidos. Por lo anterior, hemos efectuado este estudio comparando la calidad de conservación de nodos linfáticos hemales fijados con formaldehido y glutaraldehido. Los resultados muestran que ambos fijadores conservaron adecuadamente la morfología de los componentes de los nodos linfáticos hemales, sin embargo, el glutaraldehido conservó en mejores condiciones, principalmente, las células del parénquima, pero el formaldehido conservó mejor las fibras del estroma en nodos linfáticos.

Actividad hemolítica de la ortovainillina y la isovainillina sobre eritrocitos humanos / Haemolytic activity of orthovanillin and isovanillin on human erythrocytes

Los eritrocitos portadores de hemoglobina S ( b 6 glu ® val ), son menos flexibles que los eritrocitos normales, lo que los hace más frágiles y se hemolizan con mayor facilidad. La ortovainillina y la isovainillina, isómeros químicos de la vainillina, pueden inhibir la polimerización de la desoxihemoglobina S (actividad antipolimerizante) y evitar la falciformación de los eritrocitos. Se determinó la actividad citotóxica de estos compuestos sobre eritrocitos normales y SS, a razones molares 1:1, 1:4, 1:8 y 1:10, por espectrofotometría midiendo la absorbancia a una longitud de onda l =545 nm de la hemoglobina libre en el sobrenadante, después de incubar la solución de eritrocitos con los compuestos, y se determinó el porcentaje de hemólisis. Los resultados muestran que el porcentaje de hemólisis promedio calculado fue inferior al 1 por ciento. No se detectaron diferencias estadísticamente significativas entre las medias por razón molar en una misma clase de eritrocitos (p=0,05) ni una dependencia entre la concentración y la actividad hemolítica. Se compararon las medias entre ambos tipos de eritrocitos, para todas las relaciones molares, y no se observaron diferencias estadísticamente significativas. Se compararon, además, los resultados de trabajos anteriores sobre la actividad hemolítica de la vainillina con la de sus isómeros estructurales, y se obtuvo que la isovainillina y la ortovainillina provocaron porcentajes de hemólisis inferiores a los provocados por la vainillina. La baja actividad hemolítica de estos aldehídos aromáticos potencia su actividad antipolimerizante

Estudio cualitativo de relación estructura-actividad de aldehídos aromáticos, potenciales agentes antisickling / Qualitative study of the structure-activity relationship of aromatic aldehydes, potential antisickling agents

Se realizó un estudio cualitativo de relación estructura-actividad (SAR) con 27 aldehídos aromáticos de interés como potenciales agentes antisickling. Los datos de actividad: modificación de la HbA intracelular (A1) y efectos sobre la afinidad por el oxígeno (A2), se tomaron de estudios publicados en la literatura. Se emplearon descriptores mecanocuánticos determinados mediante cálculos semiempíricos (MNDO/PM3 y CNDO). Los resultados obtenidos sugieren nuevas consideraciones sobre el papel de diversos parámetros moleculares en la actividad de estos compuestos. La baja polaridad del grupo carbonilo favorece la actividad A1, así como la presencia de sustituyentes electroaceptores, disminuidores de esta polaridad. El papel de los sustituyentes y el anillo no se reduce al de moduladores de la polaridad del carbonilo. Las evidencias encontradas podrían ser útiles en trabajos de síntesis orientados a obtener compuestos con mejores perfiles de actividad antisickling

Reactions with spiroepoxy-alkanones. part III+ reaction of spirooctanones with dimethylamine aldehyde and then thiourea

Spirooctanones I react with dimethylamine at different conditions to give compounds II and III, and in the presence of aldehydes give IV. Compounds IV react with thiourea to give V, which undergoes cyclization with chloroacetic acid to give VI. Compounds VI condensed with dimethylamino-benzaldehyde to give VII. Some of the prepared compounds are biologically active

Spectroscopic studies of some new azino compounds

The electronic absorption spectra of some new azino compounds derived from hydrazine and different aldehyde and ketones are investigated in organic solvents of varying polarities. The important bands in the IR spectra and the main 1H NMR signals are assigned and discussed in relation to molecular structure. Although work on the absorption spectra of azino compounds has been reported in the literature [1-3], a series of new azino compounds derived from hydrazine and different aldehyde and ketones was prepared and in order to throw more light on their structures, these compounds were subjected to elemental analysis, spectroscopic studies and Uv-visible spectral behavior in organic solvents of varied polarity. The study included also the assignment of IR bands and 1H NMR signals by considering the effect of molecular structure on the spectral behavior

Use of pH studies to determine the kinetic and chemical mechanism of yeast alcohol dehydrogenase with primary aliphatic alcohols and aldehydes.

A complete list of all steady-state kinetic constants for the yeast alcohol dehydrogenase (EC 1.1.1.1) catalyzed oxidation of ethanol, propan-1-ol and butan-1-ol, and for the reduction of acetaldehyde and propionaldehyde was collected in the pH range 6-10, and an appropriate pH profile for each constant was constructed. A common minimal mechanism with all these substrates has been postulated and pKa values and the pH independent limiting values have been assigned for the rate constants.

Molecular association in binary mixtures of polyethylene glycol and some aldehydes

Dipole-dipole association in mixtures of polyethylene glycol [400] with both propionaldehyde and benzaldehyde, as representative aliphatic and aromatic aldehydes, has been studied. Decreases in molar volume were calculated from density measurements at different temperatures using the intercept method. The results indicated the formation of the associated aggregates in the ratio [2 propionaldehyde: 1 polyethylene glycol] and [1 benzaldehyde: 1 polyethylene glycol], which mean that the association between the aliphatic aldehyde and polyethylene glycol is greater than that for the aromatic aldehyde. This result is in an agreement with the thermodynamic parameters of dielectric relaxation and viscous flow; namely, the apparent activation energy of dielectric relaxation delta H epsilon and the enthalpy of activation of viscous flow delta Hv calculated from dielectric and viscosity measurements for the two systems