Influence of Dilute Acetic Acid Treatments on Survival of Monoecious Hydrilla Tubers in the Oregon House Canal, California

Hydrilla (Hydrilla verticillata (L.f.)Royle), a serious aquatic weed, reproduces through formation of underground tubers. To date, attacking this life-cycle stage has been problematic. The purpose of this study was to measure the impact of exposure to dilute acetic acid on monoecious hydrilla tubers under field conditions. In this field experiment, treatments were acetic acid concentration (0, 2.5, or 5%) and sediment condition (perforated or not perforated). Each of 60, 1x1 m plots (in the Oregon House Canal) were randomly assigned to one treatment. Two weeks after treatment, we collected three samples from each plot. One was washed over 2 mm wire mesh screens to separate tubers from sediment. Relative electrolyte leakage was measured for one tuber from each plot. Five additional tubers from each plot were placed in a growth chamber and sprouting monitored for four weeks. A second sample from each plot was placed in a plastic tub and placed in an outdoor tank, filled with water. These samples were monitored for tuber sprouting. Relative electrolyte leakage increased significantly for tubers exposed to 2.5% or 5% acetic acid. Effects on tubers in perforated sediment were reduced. Exposure to acetic acid inhibited tuber sprouting by 80 to 100%, in both chamber and outdoor tests. These results confirm findings from earlier laboratory/greenhouse experiments, and suggest that this approach may be useful in the management of hydrilla tuber banks in habitats where the water level can be lowered to expose the sediments.

The COMT Val158 Met polymorphism as an associated risk factor for Alzheimer disease and mild cognitive impairment in APOE 4 carriers

Background: The aim of this study is to examine the influence of the catechol-O-methyltranferase (COMT) gene (polymorphism Val158 Met) as a risk factor for Alzheimer's disease (AD) and mild cognitive impairment of amnesic type (MCI), and its synergistic effect with the apolipoprotein E gene (APOE). A total of 223 MCI patients, 345 AD and 253 healthy controls were analyzed. Clinical criteria and neuropsychological tests were used to establish diagnostic groups. The DNA Bank of the University of the Basque Country (UPV-EHU) (Spain) determined COMT Val158 Met and APOE genotypes using real time polymerase chain reaction (rtPCR) and polymerase chain reaction (PCR), and restriction fragment length polymorphism (RFLPs), respectively. Multinomial logistic regression models were used to determine the risk of AD and MCI. Results: Neither COMT alleles nor genotypes were independent risk factors for AD or MCI. The high activity genotypes (GG and AG) showed a synergistic effect with APOE epsilon 4 allele, increasing the risk of AD (OR = 5.96, 95% CI 2.74-12.94, p < 0.001 and OR = 6.71, 95% CI 3.36-13.41, p < 0.001 respectivily). In AD patients this effect was greater in women. In MCI patients such as synergistic effect was only found between AG and APOE epsilon 4 allele (OR = 3.21 95% CI 1.56-6.63, p = 0.02) and was greater in men (OR = 5.88 95% CI 1.69-20.42, p < 0.01). Conclusion: COMT (Val158 Met) polymorphism is not an independent risk factor for AD or MCI, but shows a synergistic effect with APOE epsilon 4 allele that proves greater in women with AD.

Recognition of double helical DNA by purine oligonucleotides via triple helix formation

Oligonucleotide-directed triple helix formation is one of the most versatile methods for the sequence specific recognition of double helical DNA. Chapter 2 describes affinity cleaving experiments carried out to assess the recognition potential for purine-rich oligonucleotides via the formation of triple helices. Purine-rich oligodeoxyribonucleotides were shown to bind specifically to purine tracts of double helical DNA in the major groove antiparallel to the purine strand of the duplex. Specificity was derived from the formation of reverse Hoogsteen G•GC, A���AT and T•AT triplets and binding was limited to mostly purine tracts. This triple helical structure was stabilized by multivalent cations, destabilized by high concentrations of monovalent cations and was insensitive to pH. A single mismatched base triplet was shown to destabilize a 15 mer triple helix by 1.0 kcal/mole at 25°C. In addition, stability appeared to be correlated to the number of G•GC triplets formed in the triple helix. This structure provides an additional framework as a basis for the design of new sequence specific DNA binding molecules.

In work described in Chapter 3, the triplet specificities and required strand orientations of two classes of DNA triple helices were combined to target double helical sequences containing all four base pairs by alternate strand triple helix formation. This allowed for the use of oligonucleotides containing only natural 3'-5' phosphodiester linkages to simultaneously bind both strands of double helical DNA in the major groove. The stabilities and structures of these alternate strand triple helices depended on whether the binding site sequence was 5'-(purine)_m (pyrimidine)_n-3' or 5'- (pyrimidine)_m (purine)_n-3'.

In Chapter 4, the ability of oligonucleotide-cerium(III) chelates to direct the transesterfication of RNA was investigated. Procedures were developed for the modification of DNA and RNA oligonucleotides with a hexadentate Schiff-base macrocyclic cerium(III) complex. In addition, oligoribonucleotides modified by covalent attachment of the metal complex through two different linker structures were prepared. The ability of these structures to direct transesterification to specific RNA phosphodiesters was assessed by gel electrophoresis. No reproducible cleavage of the RNA strand consistent with transesterification could be detected in any of these experiments.

Distribuição de tensões em próteses parciais fixas posteriores livres de metal com retentores intracoronários: análise em elementos finitos

Para reabilitar a ausência de um elemento dentário posterior, as próteses parciais fixas (PPF) com retentores intracoronários são uma alternativa aos implantes osseointegrados. O objetivo deste estudo foi avaliar a distribuição de tensões nessas próteses com três combina��ões de materiais: cerâmica de zircônia parcialmente estabilizada por ítria (ZPEI) revestida por cerâmica de fluorapatita (α), cerâmica de dissilicato de lítio (β) ou compósito fibrorreforçado (γ). Na composição α, foram analisadas a presença ou ausência da cerâmica de revestimento na parede cervical das caixas proximais e três varia��ões na área total da seção transversal dos conectores (4 mm de largura x 3,2, 4,2 ou 5,2 mm de altura). Em 8 modelos bidimensionais de elementos finitos, uma carga vertical de 500 N foi aplicada na fossa central do pôntico e as tensões principais máximas (tra��ão) e mínimas (compressão) foram apontadas em MPa. Inicialmente foram avaliados os 6 modelos com PPF de ZPEI e suas varia��ões. Os maiores valores das tensões de tra��ão foram encontrados no terço cervical dos conectores. Quando presente nestas regiões, a cerâmica de revestimento recebeu tensões acima do limite de sua resistência à flexão. Na compara��ão entre os modelos sem cerâmica de revestimento na parede cervical das caixas proximais, mesmo aquele com conectores de 3,2 x 4 mm, cuja infraestrutura apresentava 2,5 x 3 mm, poderia ser recomendado para uso clínico. Altos valores de tensões de compressão foram registrados entre o terço oclusal e médio dos conectores, correspondente à união entre as cerâmicas, o que poderia ocasionar, devido à flexão, falhas adesivas. Posteriormente, o modelo de ZPEI com a cerâmica de fluorapatita ausente da parede cervical das caixas proximais e área total dos conectores de 4,2 x 4 mm foi comparado aos dois outros materiais com conectores de mesma área. Na PPF de dissilicato de lítio, os valores representaram uma provável viola��ão do limite de sua resistência à flexão. A PPF de compósito fibrorreforçado apresentou tensões bem abaixo do limite de resistência à flexão de sua infraestrutura, mas, como no modelo de ZPEI, tensões compressivas se concentraram com alto valor entre o terço oclusal e médio dos conectores, local de união entre a resina composta e a infraestrutura de fibras. Os resultados mostraram que a cerâmica de dissilicato de lítio e a presença da cerâmica de fluorapatita na parede cervical das caixas proximais deveriam ser contraindicadas para a condição proposta. Parece viável uma área de conectores na infraestrutura de ZPEI com no mínimo 2,5 x 3 mm. A PPF de compósito fibrorreforçado apresenta resistência estrutural para a situa��ão estudada, mas, como também aquelas compostas de ZPEI, aparenta ter como pontos fracos a adesão entre a infraestrutura e o material de cobertura e a própria resistência deste último.

Bicyclo[3.2.0]hepta-1,4,6-triene: synthesis, thermal rearrangement, and anion formation

In order to determine the properties of the bicycloheptatrienyl anion (Ia) (predicted to be conjugatively stabilized by Hückel Molecular Orbital Theory) the neutral precursor, bicyclo[3. 2. 0] hepta-1, 4, 6-triene (I) was prepared by the following route.

Reaction of I with potassium-t-butoxide, potassium, or lithium dicyclohexylamide gave anion Ia in very low yield. Reprotonation of I was found to occur solely at the 1 or 5 position to give triene II, isolated as to its dimers.

A study of the acidity of I and of other conjugated hydrocarbons by means of ion cyclotron resonance spectroscopy resulted in determination of the following order of relative acidities:

H₂S ˃ C₅H₆ ˃ CH₃NO₂ ˃ 1, 4- C₅H₈ ˃ I ˃ C₂H₅OH ˃ H₂O; cyclo-C₇H₈ ˃ C₂ H₅OH; фCH₃ ˃ CH₃OH

In addition, limits for the proton affinities of the conjugate bases were determined:

350 kcal/mole ˂ PA(C₅ H₅^-) ˂ 360 kcal/mole

362 kcal/mole ˂ PA(C₅H₇^-, Ia, cyclo-C₇H₇^-) ˂ 377 kcal/mole PA(фCH₂^-) ˂ 385 kcal/mole

Gas phase kinetics of the trans-XVIII to I transformation gave the following activation parameters: E_a = 43.0 kcal/mole, log A = 15.53 and ∆S^ǂ (220°) = 9.6 cu. The results were interpreted as indicating initial 1,2 bond cleavage to give the 1,3-diradical which closed to I. Similar studies on cis-XVIII gave results consistent with a surface component to the reaction (Ea</sub> = 22.7 kcal/mole; log A = 9.23, ∆S^ǂ (119°) = -18.9 eu).</sub>

The low pressure (0.01 to 1 torr) pyrolysis of trans-XVIII gave in addition to I, fulvenallene (LV), ethynylcyclopentadiene (LVI) and heptafulvalene (LVII). The relative ratios of the C₇H₆ isomers were found to be dependent upon temperature and pressure, higher relative pressure and lower temperatures favoring formation of I. The results were found to be consistent with the intermediacy of vibrationally excited I and subsequent reaction to give LV and LVI.

Investiga��ão de muta��ões no gene ATP13A2 como um fator de risco para a Doença de Parkinson em pacientes brasileiros

A doença de Parkinson (DP) é a desordem neurodegenerativa motora mais frequente, com uma prevalência de, aproximadamente, 1% entre indivíduos com mais de 60 anos de idade, aumentando para 4 a 5% entre os indivíduos com idade superior a 85 anos. Esta condição é caracterizada pela perda seletiva dos neurônios dopaminérgicos da substância negra e pela presença de inclusões protéicas ricas em α-sinucleína nos neurônios sobreviventes. Pouco se sabe sobre a etiologia e a patogênese da DP. A maioria dos casos aparece esporadicamente, podendo estar associados a diversos fatores de risco ambientais e genéticos. Na última década, estudos de liga��ão identificaram 15 loci cromossômicos (PARK1 a PARK15) relacionados à DP e, nestes, um novo gene, ATP13A2, tem sido associado a casos de DP de início precoce. Esse gene está situado no 1p36 e codifica a proteína ATPase tipo-P da subfamília P5, de localiza��ão lisossômica, que é expressa em diversos tecidos, principalmente no cérebro. Muta��ões em ATP13A2 levam à forma��ão de proteínas truncadas que ficam retidas no reticulo endoplasmático e posteriormente são degradadas pelo proteossomo, podendo causar a disfunção proteossômica, decorrente da sobrecarga gerada pela proteína mutante, ou causar a disfunção lisossômica, ambas gerando agrega��ão tóxica. Este trabalho tem como objetivo realizar a análise molecular do gene ATP13A2 em uma amostra de 116 pacientes brasileiros com DP, de manifesta��ão precoce (<50 anos), de forma a avaliar se muta��ões neste gene representam um fator de risco para a DP. O DNA foi extra��do a partir de leucócitos do sangue periférico ou de saliva e a análise molecular dos éxons 2, 3, 12, 13, 14, 15, 16, 26 e 27, bem como, dos limites íntronéxons foi realizada por sequenciamento automático dos produtos da PCR. Identificamos oito variantes de sequência: quatro variantes intrônicas (uma no íntron 2, uma no íntron 13 e duas no íntron 27) e quatro variantes silenciosas (uma no éxon 3, 16, 26 e 27). Com base em dados da literatura e através de análises in silico e compara��ão com amostras controle, classificamos a altera��ão intrônica c.3084- 3C>T, e as altera��ões silenciosas c.2970G>A e c.3192C>T como não patogênicas; as altera��ões intrônicas c.106-30G>T, c.1306+42_1306+43 insC e c.3083+24C>T, e as altera��ões silenciosas c.132A>G e c.1610G>T foram classificadas como provavelmente não patogênicas. Nosso achados corroboram àqueles encontrados em outras popula��ões e indicam que muta��ões no gene ATP13A2 não são uma causa comum de DP na amostra de pacientes brasileiros analisados. No entanto, se faz necessário estender nossas análises para outras regiões gênicas, a fim de determinar o real papel deste gene na etiologia da DP em nossa popula��ão.

Comportamento sexual e uso de álcool e drogas entre homens que fazem sexo com homens no Ceará: tendências e práticas de risco para DST/AIDS

No Brasil, o s casos de AIDS entre homens que fazem sexo com homens (HSH) predominaram durante um longo período. A partir da década de 90, observa-se um declínio nesta categoria com o aumento de casos entre heterossexuais. Na região Nordeste, entretanto, os casos de AIDS entre HSH representam, ainda, cerca de 50% do total dos casos registrados em anos recentes. Nosso objetivo foi estudar o comportamento sexual e o padrão de consumo de drogas e álcool entre HSH no Ceará, enfatizando as tendências recentes e suas rela��ões com práticas sexuais de risco para DTS/AIDS. Foram realizados quatro estudos seccionais em 1995, 1998, 2002 e 2005 no Ceará, nordeste do Brasil. A popula��ão do estudo foi composta por homens que fazem sexo com homens (HSH), com 14 anos ou mais , que referiram prática sexual anal ou oral com homens nos últimos 12 meses. A seleção dos participantes utilizou técnicas do tipo Snow Ball (1995, 1998, 2002); Time Space Sampling (2002) e Respondent Driven Sampling (2005). O primeiro artigo enfoca as tendências do comportamento sexual em Fortaleza ao longo destes quatro períodos e o segundo os preditores do consumo de álcool e drogas nos municípios de Fortaleza (n=401), Sobral (n=100) e a região do Cariri (n=100) em 2002. Análise se basearam nas compara��ões entre proporções, utilizando o teste do de Pearson e intervalos de 95% de confiança (IC95%) e análise de regressão logística multivariada para avalia��ão dos fatores associados ao consumo de álcool e drogas, utilizando-se como medida de associa��ão a razão de chances (odds ratio OR) e seus respectivos intervalos de 95% de confiança. Resultados Práticas sexuais: Elevado percentual da popula��ão estudada referiu práticas sexuais de risco em 1995 (49,9%), decrescendo significativamente em 1998 (32,6%), tornando a crescer em 2002 (54,6%) e apresentando os menores percentuais em 2005 (31,4%). Este padrão não apresentou grandes varia��ões por idade, mas em rela��ão à escolaridade observou-se que os indivíduos com escolaridade mais elevada aumentaram as práticas de risco entre 1998 (28,6%) e 2002 (46,5%) decrescendo no último período (21,0%) enquanto aqueles com baixa ou média escolaridade só mostraram uma queda significativa no comportamento de risco entre 2002 (82,1% - baixa; 67,7% - média) e 2005 (29,1% - baixa; 34,3 média). A prática sexual anal com preservativo cresceu no decorrer dos anos variando de 43,3% a 53,7% entre a primeira e a última onda ( de tendência p<0.001). A rela��ão anal sem preservativo foi uma prática com alto percentual na maioria dos anos. De 2002 a 2005, houve uma diminuição significativa (de alto percentual na maioria dos anos. De 2002 a 2005, houve uma diminuição significativa (de 57,7% para 26,3%) das rela��ões fixas monogâmicas. Consumo de álcool e drogas: No estudo, 63% dos HSH participantes foram classificados como bebedores que se embriagam. Observou-se que o consumo crescente de álcool leva a um aumento do uso concomitante de outras drogas, sejam lícitas ou ilícitas. Foram variáveis preditoras de beber se embriagando: ter de 21 a 30 anos (OR: 1,5; IC 95%: 1,1-2,9); ter mais que 30 anos (OR: 1,6: IC95%: 1,2-2,3); ser solteiro/separado/divorciado (OR:3,0%; IC95%: 1,7-5,3); ser da ra��a negra (OR: 2,0 IC95%: 1,7-2,01); ser da ra��a parda (OR: 1,8 IC95%: 1,3-2,6); receber dinheiro por sexo (OR:2,0 IC95%: 1,8-2,9). As práticas sexuais dos SHS em Fortaleza apresentaram varia��ões significativas ao longo doa anos estudados, semelhantemente a outros estudos internacionais. Vários fatores poderiam ser responsáveis por explicar o comportamento da curva observada em Fortaleza, seja no âmbito local, nacional ou internacional. Entre os fatores que podem explicar altera��ões observadas estariam: 1) redução nos recursos destinados à prevenção da AIDS no pa��s devido a retirada de alguns organismos de coopera��ão internacional que se voltaram para outros pa��ses, como na África Leste Europeu, levando o Brasil a priorizar segmentos populacionais com maior vulnerabilidade; 2) grande impacto na prevenção das DST /AIDS na comunidade de homo/bissexuais masculinos, especialmente nos anos de 1998 a 2002; 3) o avanço no tratamento, surgimento de novas drogas, melhora da qualidade de vida e aumento da sobrevida, contribuindo para a construção da falsa ideia de segurança na popula��ão. Neste estudo a escolaridade mostrou-se um fator importante associado ao envolvimento em práticas sexuais não seguras. Os indivíduos com mais baixa escolaridade, no período de 1995 a 2002, se envolveram em maior risco, aparentando não terem sido atingidos pelas campanhas que possam ter ocorrido, principalmente no período de 1995 a 1998. A maior escolaridade apresenta-se como fator de proteção em todo o período estudado, provavelmente pelo maior acesso à informa��ão. Finalmente, pode-se observar no ano de 2002 um elevado percentual de homens que consomem cinco ou mais doses em um dia típico e associam outras drogas ao consumo do álcool. Tal comportamento, dentro da popula��ão HSH, embora não seja caracterizado como dependência química, é alterado de maneira significativa pelo efeito etílico, levando à outras práticas de risco. Também se observou em nosso estudo que o consumo crescente de álcool leva a um aumento do uso de outras drogas, atuando para a adoção de comportamentos de risco. Existem evidências que suportam rela��ão entre uso de outras drogas e a prática sexual de risco. Os indivíduos que referiram receber dinheiro em troca de sexo foram mais frequentemente classificados como bebedores que se embriagam. Os achados deste estudo mostram a importância da realiza��ão de uma vigilância comportamental contínua em rela��ão ao HIV favorecendo o entendimento da dinâmica da epidemia junto das DST/AIDS nesta popula��ão vulnerável, assim como a importância que o álcool assume como problema de sa��de pública neta popula��ão específica e a necessidade de se direcionar medidas voltadas para a sua prevenção.

Aspectos clínico-epidemiológicos e microbiológicos de processos infecciosos causados por Corynebacterium spp em pacientes de centro de referência oncológico Rio de Janeiro, Brasil

Os resultados permitiram a reda��ão de quatro artigos. Aspectos microbiológicos e clínicos de corinebacterioses em pacientes com câncer observados durante cinco anos foram descritos no Artigo 1. No Artigo 2 foram apresentados casos de bacteremia causados por corinebactérias invasivas não toxigênicas em dois períodos com intervalo de sete anos. As infecções em pacientes com câncer por C. diphtheriae, causando casos clínicos atípicos foram descritas no Artigo 3, além do estudo dos principais fatores de virulência de uma cepa de C. diphtheriae isolada de infecção associada ao cateter de nefrostomia foi descrita no Artigo 4. Resumidamente no Artigo 1, além dos aspectos clínico-epidemiológicos foram avaliados os perfis de resistência aos antimicrobianos e o potencial de virulência dos micro-organismos. Em cinco anos, 932 amostras de corinebactérias, com perfis de resistência aos antimicrobianos testados, foram isoladas de pacientes com câncer. As espécies predominantes foram Corynebacterium amycolatum (44,7%), Corynebacterium minutissimum (18,3%) e Corynebacterium pseudodiphtheriticum (8,5%). O uso de catéteres de longa permanência e a neutropenia, foram às condições importantes para infecção por corinebactérias. As doenças de base mais comuns foram os tumores sólidos. Pacientes hospitalizados apresentaram risco seis vezes maior de morrer, quando relacionadas às taxas de mortalidade com 30 dias (RC= 5,5; IC 95%= 1,15-26,30; p= 0,033). As bacteremias (Artigo 2) causadas por corinebactérias foram observadas em dois períodos: 2003-2004 (n=38) e de 2012-2013 (n=24). As espécies multirresistentes C. amycolatum e Corynebacterium jeikeium foram os principais responsáveis pelos quadros de bacteremia. Havia 34 pacientes com tumores sólidos e 28 pacientes com doenças linfoproliferativas, sendo que 21 deles apresentavam neutropenia e 54 utilizavam cateter venoso central. Em 41 pacientes havia infecção relacionada ou associada aos dispositivos intravasculares. Os pacientes com bacteremia responderam ao tratamento com vancomicina após a remoção do cateter. O comportamento agressivo da neoplasia, o tempo de interna��ão hospitalar e o uso de CVC aumentaram o risco de bacteremias por Corynebacterium spp. No Artigo 3, 17 casos de infecções atípicas causadas por Corynebacterium diphtheriae foram diagnosticadas de 1996 a 2013. A incidência de C. diphtheriae correspondeu a 15,8 casos/100.000 admissões, 465 vezes maior que a incidência de difteria na popula��ão brasileira. Sintomas toxêmicos foram observados em nove pacientes, embora quadros de difteria clássica e endocardite não fossem observados. O perfil eletroforético em campo pulsado (PFGE) demonstrou um perfil de distribuição endêmica, apesar de haver dois casos de pacientes com o mesmo perfil eletroforético sugerindo transmissão relacionada aos cuidados à sa��de. A adesão em superfícies bióticas e abióticas e produção de biofilme em cateter de poliuretano (Artigo 4) foi demonstrada em C. diphtheriae não toxigênico no sítio de inserção do cateter de nefrostomia. Os dados desses artigos permitiram concluir que (i) diferentes espécies de corinebactérias multirresistentes foram capazes de causar infecções em pacientes com câncer, incluindo bacteremias; (ii) C. diphtheriae foi capaz de causar infecções graves em indivíduos imunocomprometidos, incluindo infecções relacionadas ao uso de dispositivos invasivos em popula��ões de risco, tais como pacientes com câncer.

Effect of adrenalectomy and corticosterone replacement on prepulse inhibition and locomotor activity in mice

1 Stress is a risk factor in psychiatric illnesses such as schizophrenia. The aim of the present study was to investigate the effect of different circulating levels of the adrenal steroid corticosterone (CORT) on locomotor hyperactivity and prepulse inhibition of acoustic startle, two behavioural animal models of aspects of schizophrenia. 2 Male C57BL/6J mice (n = 10 per group) were anaesthetised with isoflurane and sham-operated or adrenalectomised (ADX). ADX mice were implanted with 50 mg pellets consisting of 100% cholesterol, or 2, 10 or 50 mg of CORT mixed with cholesterol. CORT pellet implantation dose dependently increased plasma CORT levels 3 weeks after surgery. Starting 1 week after surgery, mice were tested for prepulse inhibition after injection of saline or 5 mg kg(-1) of haloperidol. 3 In intact mice and in mice implanted with 10 mg of CORT, haloperidol treatment significantly increased prepulse inhibition (average values from 38 - 42 to 52%). Similar results were observed when testing the mice for amphetamine-induced locomotor hyperactivity (5 mg kg(-1)). In contrast, there was no significant effect of haloperidol in mice implanted either with cholesterol or 2 or 50 mg of CORT. 4 These results in behavioural animal models of schizophrenia suggest an important role of the stress hormone CORT in modulating dopaminergic activity in this illness.

Shrinkage of Gobiocypris rarus, Procypris rabaudi, and Sinilabeo rendahli preserved in formalin

Length measurements of preserved fishes are necessary in many types of fish surveys because logistics often do not allow for fish measurement immediately after catch. If the fixative causes significant shrinkage, then the preserved lengths cannot be directly used to indicate accurate live lengths. The objective of this study was to determine how preservation in formalin affects standard length of Gobiocypris rarus larvae (24-day-old and newly hatched), larval Procypris rabaudi (4-day-old), and larval Sinilabeo rendahli (12-day-old). Fishes were measured (to nearest 0.01 mm) and individually fixed in the appropriate formalin solution (2.5% or 5.0% formalin), then re-measured at 0.5, 1, 3, 7, 14, 30, 45 and 75 days after preservation to follow the time course of shrinkage. Most of the shrinkage occurred within the first half day after preservation. The 5.0% formalin caused a higher relative shrinkage rate than did the 2.5% solution; however, the difference was not statistically significant. In G. rarus, initial shrinkage of newly hatched larvae was higher than that of 24-day-old larvae.

Optimization by orthogonal array design and humoral immunity of the bivalent vaccine against Aeromonas hydrophila and Vibrio fluvialis infection in crucian carp (Carassius auratus L.)

Aeromonas hydrophila and Vibrio fluvialis are the causative agents of a serious haemorrhagic septicaemia that affects a wide range of freshwater fish in China. In order to develop a bivalent anti-A. hydrophila and anti-V. fluvialis formalin-killed vaccine to prevent this disease, an orthogonal array design (OAD) method was used to optimize the production conditions, using three factors, each having three levels. The effects of these factors and levels on the relative per cent survival for crucian carp were quantitatively evaluated by analysis of variance. The final optimized formulation was established. The data showed that inactivation temperature had a significant effect on the potency of vaccine, but formalin concentration did not. The bivalent vaccine could elicit a strong humoral response in crucian carp (Carassius auratus L.) against both A. hydrophila and V. fluvialis simultaneously, which peaked at 3 or 5 weeks respectively. Antibody titres remained high until week 12, the end of the experiment, after a single intraperitoneal injection. The verification experiment confirmed that an optimized preparation could provide protection for fish at least against A. hydrophila infection, and did perform better than the non-optimized vaccine judged by the antibody levels and protection rate, suggesting that OAD is of value in the development of improved vaccine formulations.

小麦胚乳蛋白组分和Glu-B3位点基因核心编码区差异与小麦品质关系的研究

小麦加工品质改良已成为我国小麦育种的主要目标之一。特别是我国加入WTO以后，对小麦产品的质量提出了更高的要求，小麦品质改良的任务将更加艰巨和重要，小麦胚乳蛋白是影响小麦加工品质性状的重要因素。因此，深入了解小麦胚乳蛋白对加工品质性状的影响及其分子基础，为品质改良提供理论依据和科学指导，对加速我国小麦品质育种和优质小麦生产具有重要意义。本研究选用在麦谷蛋白5个基因位点(Glu-A1、Glu-B1、Glu-D1、Glu-B3和Glu-D3)上均含不同等位基因的小麦品种99G45和京771及Pm97034和京771杂交F9代共164个麦谷蛋白纯合系，及228个中国推广普通小麦品种和高代育成品系为试材，研究了麦谷蛋白Glu-1和Glu-3位点基因等位变异对籽粒蛋白、湿面筋含量、Zeleny沉降值和SDS沉降值间的关系；本研究还利用小麦A���B和D基因组中低分子量麦谷蛋白亚基（LMW-GS）基因特异引物，通过PCR方法克隆了1个Glu-A3位点和3个Glu-B3位点LMW-GS基因片段，在此基础上分析了不同等位基因对品质造成差异的分子基础；另外，本研究对中国近年推广的部分品种和育成的高代品系资源的多样性进行了分析。现将主要研究结果简述如下： 1. 对来自三个麦区的148份材料的醇溶蛋白组成进行了分析，结果表明，各麦区醇溶蛋白模式具有较大差异。在ω区,A7、B、E、F、G、J、P、Q、S和U仅存在于西南秋播麦区；A3、M、N、R、W和X仅存在于黄淮特种麦区；K仅存在于北方冬麦区；A6是北方冬麦区出现频率最高的带型模式，而西南秋播麦区中D出现的频率最高。ω-区的E、H和M几种模式是以前国内外未曾报道的。且初步确定，这些模式对品质性状具有正效应。至于γ区,A���B、D、E和F在各区均有出现，其中B和E在各区出现的频率都很高，在26.1-39.6％之间。相反，H 仅出现在黄淮特种麦区，J仅限于西南秋播麦区。对于β-区醇溶蛋白，B型模式在所有区中都相当高，而模式A���存在于第三区.对于α-区，模式A���Ⅲ区而模式D在Ⅱ区出现的频率很高。1BL.1RS易位系在中国小麦品种中出现频率高达41.2％，在I, II和Ⅲ麦区的出现频率分别为 45.5、43.5和35.2%。各生态区模式的差异可能是品种适应不同生态条件和人为选择的结果，但这有待进一步证明。由于醇溶蛋白位点（Gli-1）与LMW-GS位点（Glu-3）紧密连锁，本结果可为下面确定普通小麦LMW-GS等位基因变异所用。 2. 利用Gli-1与Glu-3的紧密连锁，以228个小麦品种/系为材料，首次对中国小麦品种麦谷蛋白亚基的6个位点进行综合分析，研究小麦籽粒蛋白与品质性状间的关系，结果表明6个高分子量（HMW）和低分子量（LMW）麦谷蛋白位点对蛋白质含量的效应大小为，Glu-D1>Glu-B3>Glu-A1=Glu-B1> Glu-A3=Glu-D3；对GMP含量的效应大小为, Glu-A3>Glu-B3>Glu-D1> Glu-B1>Glu-A1>Glu-D3；对湿面筋含量的效应大小为, Glu-B1>Glu-B3= Glu-D3>Glu-A3>Glu-A1>Glu-D1；对Zeleny沉降值的效应大小为, Glu-A1> Glu-B3>Glu-D3>Glu-D1>Glu-B1>Glu-A3；对SDS沉降值的效应大小为, Glu-B3>Glu-A1=Glu-D1=Glu-A3>Glu-D3>Glu-B1。对蛋白含量而言，各位点的最佳组合方式为1、17+18、5+10、Glu-A3e、Glu-B3g、Glu-D3b；对湿面筋含量而言，各位点的最佳组合方式为1、6+8、5+10、Glu-A3d、Glu-B3c、Glu-D3b；对Zeleny沉降值而言，各位点的最佳组合方式为N、17+18、5+10、Glu-A3d、Glu-B3d、Glu-D3b；对SDS沉降值而言，各位点的最佳组合方式为1、7+8、2.2+12、Glu-A3b、Glu-B3g、Glu-D3b。另外，分析了稀有亚基对5+12与2.2+12与品质性状的关系，认为5+12对品质有负效应，2.2+12对品质有正效应。在品质育种时，应对优异组合或优异亚基加以利用。 3. 首次利用重组自交系（RILs）为材料，研究麦谷蛋白亚基表达量与品质性状的关系，通过对重组自交系中各HMW-GS表达量的分析，认为，就单个亚基的表达量而言，7亚基最高；其次为2亚基、5亚基、12亚基和10亚基；亚基9和1的表达量最小；N亚基不表达。对成对出现的亚基对而言，x型和y型亚基的总表达量2+12>5+10>7+9>17+18。就单个亚基与品质性状的关系而言，仅有10亚基的表达量与蛋白含量的相关性达5%的显著水平，2亚基的表达量与湿面筋含量呈负相关，显著水平也达5%，其余单个亚基对品质性状均无显著影响；就x型/y型亚基的比例来看，2/12和5/10对湿面筋含量都有显著的负效应；对某一位点等位基因控制的亚基表达总量来看，2+12对SDS沉降值有显著负效应。另外，本研究得出：2＋12的亚基对的负效应主要体现在2亚基上，且在同一位点上，x型亚基的表达量大于y型。所以推导稀有亚基组合2+10很可能也是劣质亚基。 4. 以 Glu-A1、Glu-B1、Glu-D1、Glu-B3和Glu-D3作为5个因素对99G45/京771和Pm97034/京771杂交后代的蛋白质含量和SDS沉降值进行多因素方差分析。结果表明，Glu-A1和Glu-D3对蛋白含量的加性效应达5％显著水平；Glu-D1 * Glu-D3对蛋白质含量的互作效应也达5%显著水平；其余位点的加性和互作效应对蛋白质含量的影响均不显著。对SDS 沉降值而言，Glu-D1的加性效应最大,贡献率为4.2 % ,达1 %显著水平,其次是Glu-B1位点,贡献率为3.3% ,达5%显著水平。其余位点对SDS 沉降值的加性和互作效应均未达5%显著水平。总体而言, 各位点对蛋白含量的效应大小为Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3；对SDS沉降值的效应大小为Glu-D1>Glu-B1> Glu-D3>Glu-A1> Glu-B3。Glu-D1和Glu-D3位点上等位基因变异对蛋白含量有显著或极显著影响，含Glu-D1d和Glu-D3 GD、Glu-D3 JD基因的株系分别比含Glu-D1a���Glu-D3 PD基因的株系有较高的蛋白含量；在该遗传背景下，麦谷蛋白各基因位点对蛋白含量的效应大小依次排列为：Glu-A1位点1>N；Glu-B1位点7+9>17+18>14+15；Glu-D1位点5+10>2+12；Glu-B3位点GB>JB>PB；Glu-D3位点GB>JB>PB。对SDS沉降值的效应大小依次排列为：Glu-A1位点1>N；Glu-B1位点7+9=17+18>14+15；Glu-D1位点5+10>2+12；Glu-B3位点GB>JB>PB；Glu-D3位点GB>JB>PB。所以，对蛋白含量和SDS沉降值均较好的组合为1，7+9，5+10，GB，GD。 5. 因为GB和PB对品质的效应有显著差异，选取LMW-GS位点特异扩增引物对京771、99G45和Pm97034的Glu-B3位点进行扩增，结果得到三个不一样的扩增片段（Genebank号为DQ539657－DQ539659），得到的基因片段与Genebank中已报道的同类序列高度同源。通过克隆片段组成的分析，发现对Pm97034的序列较京771和99G45段少一个7氨基酸的重复单元，这可能是它较另外两个片段对面筋强度影响小的主要原因；另外，在99G45的序列中，124位处出现L（亮氨酸）代替P（脯氨酸），158位处出现了T（苏氨酸）代换M(蛋氨酸)，这可能是99G45Glu-B3位点序列对SDS沉降值的效应显著优于Pm97034的原因。 6．通过对RILs各位点同普通小麦品种（系）各位点与品质关系的比较，发现对SDS沉降值的效应，各位点在不同研究材料中是不同的，普通小麦中：Glu-B3>Glu-A1=Glu-D1=Glu-A3>Glu-D3>Glu-B1，RILs中：Glu-D1>Glu-B1> Glu-D3>Glu-A1> Glu-B3。利用重组自交系材料（完全排除了1BL/1RS易位干扰）所得到的结果与Gupta and MacRitchie (1994)所得结论一致。进一步证实了1BL/1RS易位对小麦品质的重要影响。对蛋白含量而言，普通小麦品种（系）中，Glu-D1>Glu-B3>Glu-A1=Glu-B1> Glu-A3=Glu-D3，RILs中，Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3，和对SDS沉降值的效应一样，推断在非1BL/1RS易位的情况下，各位点对其效应应为Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3。 对同一位点的等位基因而言，普通小麦和重组自交系中Glu-A1和Glu-D1上的等位基因对品质性状的贡献是一致的，但Glu-B1上的等位基因对SDS沉降值的贡献发生了变化，普通小麦中17+18>7+9，RILs中7+9>17+18，这可能也是1BL/1RS造成的。 Baking quality improved is one of the main object of wheat bread in China. The overall objective of the present studies was to increase the understanding about protein quality in wheat, i.e. to make it possible to improve the production of wheat with desired quality for different end-uses. With the analysis of gluten protein in RILs, 99G45/Jing 771 and Pm97034/Jing, and 228 wheat cultivars or lines in China, the correlations between glutenin compositions and protein content, glutenin macropolymer(GMP), wet gluten content, Zeleny sedimentation value and SDS sedimentation value contentand breadmaking quality were studied. Also a rapid and efficient detection method of geneticpolymorphism at Glu-B3 loci in wheat was established using polymerase chain reaction(PCR).The results obtained were as follows: 1. Cultivated Chinese wheat germplasm has been a valuable genetic resource in international plant breeding. Patterns of gliadin among cultivated Chinese accessions are unknown, despite the proven value and potential novelty. The objective of this work was to analyse the diversity within improved Chinese wheat germplasm. The electrophoretic banding patterns of gliadin in common wheat cultivars and advanced lines were determined by acid-polyacrylamide gel electrophoresis. For 148 leading commercial cultivars and promising advanced lines used in our study, 48 patterns were identified, 29 corresponding to ω-gliadin, 9 to γ-gliadin, 5 to β-gliadin and 5 to α-gliadin. The most frequent patterns were A6 in ω; B in γ; B in β and A in the region of α. 116 band types appeared in the148 samples: 94 accessions had unique gliadin types, and 22 gliadin types while not unique were found in 54 accessions. The gliadin patterns of Chinese wheat cultivars and lines greatly differed from the patterns of wheat lines from other countries. Three patterns, E, J, H, M, N and O in the ω-zone had not previously been reported. Three wheat zones，the Northern Winter Wheat Region, the Yellow and Huai Valley River valleys Winter Wheat Region and the Southwestern Winter Wheat Region，in China showed different frequencies in their gliadin patterns. This information can be used to monitor genetic diversity with Chinese wheat germplasm. 2. To analyse the relationship between the loci and characteristics quality, we utilized the 228 cultivars/lines. The results showed that : For protein content, Glu-D1 >Glu-B3>Glu-A1=Glu-B1>Glu-A3=Glu-D3. For GMP content, Glu-A3>Glu-B3 >Glu-D1>Glu-B1>Glu-A1>Glu-D3. For wet gluten content, Glu-B1>Glu-B3= Glu-D3>Glu-A3>Glu-A1>Glu-D1. For Zeleny sedimentation value, Glu-A1>Glu-B3> Glu-D3>Glu-D1>Glu-B1>Glu-A3, For SDS sedimentation value, Glu-B3>Glu-A1= Glu-D1= lu-A3>Glu-D3>Glu-B1。For protein content, the best combination of 6 loci is (1,17+18,5+10,Glu-A3e, Glu-B3g,Glu-D3b). For wet gluten content, the best combination of 6 loci is (1,6+8,5+10,Glu-A3d,Glu-B3c,Glu-D3b). For Zeleny sedimentation value, the best combination of 6 loci is (N,17+18,5+10,Glu-A3d, Glu-B3d, Glu-D3b). For SDS sedimentation value, the best combination of 6 loci is(7+8,2.2+12,Glu-A3b, Glu-B3g,Glu-D3b)。Additional, we analysed the relationship between the subunits 5+12 and 2.2+12, think that 5+12 was negative for quality, 2.2+12 is postive for quality. It should be effective utilized. 3. It’s the first time to utilize RILs to study the relationship between subunits expression quantity and characteristics quality. The results showed that: For single subunit, the expression quantity of 7 is the highest. Then the 2, 5, 12 and 10. The expression of subunit 9 and 1 is the lowest. Subunit N is not expressed. For subunits, the expression quantity of x type and y type are 2+12>5+10>7+9>17+18. The significant relation of 5% only showed between the expression quantity of subunit 10 and protein content. The relationship between expression quantity of others and characteristic quality was not significant. For x type/ytype, 2/12 and 5/10 is negative relation insignificant level. For the subunit(s) in a loci, Only 2+12 effect SDS sedimentation value negative in significant level. 4. With RILs 99G45/Jing 771 and Pm97034/Jing 771, we found that: The effective of Glu-A1, Glu-D3 and Glu-D1 * Glu-D3 for protein content is significant at 5% level. The effect of other loci for protein wre not significant. For SDS sedimentation value, the effect of Glu-D1is the highest, which contribution is 4.2 % .Then the Glu-B1, contribution is 3.3%. The effect of other loci for SDS sedimentationvalue were not significant. In total, for protein content: Glu-D3 > Glu-A1 > Glu-D1>Glu-B1>Glu-B3; for SDS sedimentationvalue: Glu-D1>Glu-B1> Glu-D3>Glu-A1>Glu-B3. The effect of alleles in Glu-D1 and Glu-D3 loci are significant at 1% or 5%. In Glu-A1, 1>N; Glu-B1, 7+9>17+18>14+15; Glu-D, 5+10>2+12; Glu-B3, GB>JB>PB; Glu-D3, GB>JB>PB. For SDS sedimentation, Glu-A1, 1>N; Glu-B1, 7+9=17+18>14+15; Glu-D1, 5+10>2+12; Glu-B3, GB>JB>PB; Glu-D3, GB>JB>PB. The best combinations for SDS sedimentation value is 1，7+9，5+10，GB，GD. 5. Because of the difference of GB and PB for SDS sedimentation value, we selected the specific primer for LMW-GS loci to amplified the Glu-B3 of Jing771, 99G45and Pm97034. We got 3 amplify fragment (Gene Bank accession number are DQ539657－DQ539659）. We found that the fragment of Pm97034 were deleted a repetitive 7 amino acid domain, which is perhaps the reason effect the gluten strength. Furthermore, in the position 124 of sequence 99G45, L has been replaced with P. Position 158, T replaced M, which may be the reason why the Glu-B3 locus of 99G45 is prefer to Pm97034 when refer to SDS sedimentation value. 6. Comparing the results of RILs and common wheat, we found that perhaps just the1BL/1RS made the difference of loci in different accession.

Investigation of PL properties of C-doped SiO2/Si samples after high energy Pb ion irradiation

Amorphous SiO2 thin films with about 400-500 nm in thickness were thermally grown on single crystalline silicon. These SiO2/Si samples were firstly implanted at room temperature (RT) with 100 keV carbon ions to 2.0 x 10(17),5.0 X 10(17) or 1.2 x 10(18) ions/cm(2), then irradiated at RT by 853 MeV Pb ions to 5.0 x 10(11), 1.0 X.10(12) 2.0 x 10(12) or 5.0 x 10(12) ions/cm(2), respectively. The variation of photoluminescence (PL) properties of these samples was analyzed at RT using a fluorescent spectroscopy. The obtained results showed that Pb-ion irradiations led to significant changes of the PL properties of the carbon ion implanted SiO2 films. For examples, 5.0 x 10(12) Pb-ions/cm(2) irradiation produced huge blue and green light-emitters in 2.0 x 10(17) C-ions/cm(2) implanted samples, which resulted in the appearance of two intense PL peaks at about 2.64 and 2.19 eV. For 5.0 x 10(17) carbon-ions/cm(2) implanted samples, 2.0 x 10(12) Pb-ions/cm(2) irradiation could induce the formation of a strong and wide violet band at about 2.90 eV, whereas 5.0 x 10(12) Pb-ionS/cm(2) irradiation could,create double peaks of light emissions at about 2.23 and 2.83 eV. There is no observable PL peak in the 1.2 x 10(18) carbon-ions/cm(2) implanted samples whether it was irradiated with Pb ions or not. All these results implied that special light emitters could be achieved by using proper ion implantation and irradiation conditions, and it will be very useful for the synthesis of new type Of SiO2-based light-emission materials.

Facile, Efficient Copolymerization of Ethylene with Bicyclic, Non-Conjugated Dienes by Titanium Complexes Bearing Bis(beta-Enaminoketonato) Ligands

Copolymerizations of ethylene with 5-vinyl-2-norbornene or 5-ethylidene-2-norbornene under the action of various titanium complexes bearing bis(beta-enaminoketonato) chelate ligands of the type, [(RN)-N-1=C(R-2)CH=C(R-3)O](2)TiCl2 (1, R-1=Ph, R-2=CF3, R-3=Ph; 2, R-1=C6H4F-p, R-2=CF3, R-3=Ph; 3, R-1=Ph, R-2=CF3, R-3=t-Bu; 4, R-1=C6H4F-p, R-2=CF3, R-3=t-Bu; 5, R-1=Ph, R-2=CH3, R-3=CF3; 6, R-1=C6H4F-p, R-2=CH3 R-3=CF3), have been shown to occur with the regioselective insertion of the endocyclic double bond of the monomer into the copolymer chain, leaving the exocyclic vinyl double bond as a pendant unsaturation. The ligand modification strongly affects the copolymerization behaviour. High catalytic activities and efficient co-monomer incorporation can be easily obtained by optimizing the catalyst structures and polymerization conditions.

7-Amino actinomycin D bound to single-stranded hairpin DNA enhanced by loop sequence-dependent luminescent Eu3+ and Tb3+ binding

Lanthanide Eu3+ and Tb3+ ions have been widely used in luminescent resonance energy transfer (LRET) for bioassays to study metal binding microenvironments. We report here that Eu3+ or Tb3+ can increase the binding affinity of antitumor antibiotic drug agent, 7-amino actinomycin D (7AACTD), binding to 5'-GT/TG-5' or 5'-GA/AG-5' mismatched stem region of the single-stranded hairpin DNA. Further studies indicate that the effect of Eu3+ or Tb3+ on 7AACTD binding is related to DNA loop sequence. Our results will provide new insights into how metal ions can enhance antitumor agents binding to their targets.