Abstract Candida spp. are naturally opportunistic and can promote infections. These yeasts can form biofilm, after penetration and adhesion to the biotic or abiotic surfaces. Preexisting diseases, treatments with drugs and radiation therapy, medical procedures, and parafunctional habits favor the installation of a fungal infection. Increased resistance to the available antifungals has become a concern. Therefore, alternative methods to control them have been evaluated, including the use of plant substances. In this study, the antibiofilm effect of R. officinalis L. extract was analyzed on C. albicans, C. dubliniensis, C. glabrata, C. krusei, and C. tropicalis. A phytochemical analysis of the extract was performed. Biofilms were formed for 48 h and exposed to the different concentrations of the extract (50, 100, and 200 mg/mL) for 5 min or 24 h. The effect of the plant extract was compared to the antifungal nystatin. Rosmarinus officinalis L. extract was constituted of phenols and flavonoids, highlighting the presence of chlorogenic acid derivatives in its composition. Biofilm reductions were observed after exposure to the plant extract for both periods. The plant extract provided a reduction similar to the antifungal. Thus, R. officinalis L. extract showed antibiofilm effect on Candida spp. comparable to the nystatin.

Eosinophils are abundant in the reproductive tract, contributing to the remodeling and successful implantation of the embryo. However, the mechanisms by which eosinophils migrate into the uterus and their relationship to edema are still not entirely clear, since there are a variety of chemotactic factors that can cause migration of these cells. Therefore, to evaluate the role of CCR3 in eosinophil migration, ovariectomized C57BL/6 mice were treated with CCR3 antagonist SB 328437 and 17β-estradiol. The hypothesis that the CCR3 receptor plays an important role in eosinophil migration to the mouse uterus was confirmed, because we observed reduction in eosinophil peroxidase activity in these antagonist-treated uteruses. The antagonist also influenced uterine hypertrophy, inhibiting edema formation. Finally, histological analysis of the orcein-stained uteruses showed that the antagonist reduced eosinophil migration together with edema. These data showed that the CCR3 receptor is an important target for studies that seek to clarify the functions of these cells in uterine physiology.

RESUMO: O presente estudo teve como objetivo avaliar o potencial de deriva de glifosato durante aplicação aérea mediante o uso de plantas de arroz como sentinelas, a fim de determinar o efeito da deriva na cultura do arroz irrigado. Para isso, foi realizado um experimento em campo utilizando delineamento inteiramente casualizado com quatro repetições, avaliando diferentes distâncias [testemunha (sem aplicação), 0; 12,5; 25; 50; 75; 100; 150; 200; 300; e 400 m] a partir do local de aplicação. O experimento foi realizado nas Granjas 4 Irmãos, localizada no município de Rio Grande, RS. A dose de aplicação de glifosato foi de 1.920 g e.a. ha-1 (Roundup Transorb®, sal de isopropilamina de glifosato 480 g e.a. L-1). Para estimação da taxa de deriva nas plantas sentinelas, foi elaborada uma curva de dose-resposta, aplicando doses crescentes de glifosato em plantas de arroz e avaliando o nível de injúria. As taxas de deriva estimadas mediante o nível de injúria em plantas sentinelas foram de 14% (150 m), 13% (200 m) e 5% (400 m). Foi observado morte de plantas nas distâncias entre 0 a 50 m, enquanto em distâncias entre 75 e 150 m a taxa de sobrevivëncia foi de 25 a 50%, causando redução na produtividade. Já nas distâncias entre 200 e 400 m não houve redução na produtividade quando comparada com a testemunha, inclusive quando os níveis de injúria atingiram de 52 a 82%. Dessa forma, conclui-se que a deriva de glifosato de 5% atingiu até 400 m da faixa de aplicação. Considerando a recomendação de deriva zero, distâncias maiores que 400 m deverão ser adotadas para evitar sintomas em plantas de arroz. Sugere-se a utilização de distâncias maiores que 400 m em estudos futuros.

ABSTRACT: This study aimed to evaluate the potential for glyphosate drift during aerial application using rice plants as sentinels, aiming to determine the effect of drift on irrigated rice crops. For this purpose, a field experiment was performed using an entirely randomized design with four replicates, evaluating different distances from the site of application [control (no application), 0, 12.5, 25, 50, 75, 100, 150, 200, 300, and 400 m]. The experiment was carried out at the Granjas 4 Irmãos farm, located in the Rio Grande city, Rio Grande do Sul State, Brazil. The glyphosate dose application was 1,920 g e.a. ha-1 (Roundup Transorb®, 480 g e.a. L-1 glyphosate isopropylamine salt). A dose-response curve was developed to estimate the drift rate in sentinel plants, by applying increasing glyphosate doses in rice plants and assessing the injury level. The drift rates estimated by the injury level in sentinel plants were 14% (150 m), 13% (200 m), and 5% (400 m). Death of the experimental units was observed for distances between 0 and 50 m, while in distances between 75 and 150 m, 25 to 50% of the plants survived, reducing productivity. In the distances between 200 and 400 m, there was no reduction in productivity when compared to the control, even when the injury levels reached 52 to 82%. Thus, we concluded that a 5% glyphosate drift reached up to 400 m from the application range. Considering the recommendation of zero drift, distances greater than 400 m should be adopted to avoid symptoms in rice plants. We suggest using distances of more than 400 m in future studies.

The heptapeptide Bj-PRO-7a, isolated and identified from Bothrops jararaca (Bj) venom, produces antihypertensive and other cardiovascular effects that are independent on angiotensin converting enzyme inhibition, possibly relying on cholinergic muscarinic receptors subtype 1 (M1R). However, whether Bj-PRO-7a acts upon the central nervous system and modifies behavior is yet to be determined. Therefore, the aims of this study were: i) to assess the effects of acute administration of Bj-PRO-7a upon behavior; ii) to reveal mechanisms involved in the effects of Bj-PRO-7a upon locomotion/exploration, anxiety, and depression-like behaviors. For this purpose, adult male Wistar (WT, wild type) and spontaneous hypertensive rats (SHR) received intraperitoneal injections of vehicle (0.9% NaCl), diazepam (2 mg/kg), imipramine (15 mg/kg), Bj-PRO-7a (71, 213 or 426 nmol/kg), pirenzepine (852 nmol/kg), α-methyl-DL-tyrosine (200 mg/kg), or chlorpromazine (2 mg/kg), and underwent elevated plus maze, open field, and forced swimming tests. The heptapeptide promoted anxiolytic and antidepressant-like effects and increased locomotion/exploration. These effects of Bj-PRO-7a seem to be dependent on M1R activation and dopaminergic receptors and rely on catecholaminergic pathways.

Arthritis is positively associated with the decline of sex hormones, especially estrogen. Tamoxifen (TMX) is a selective estrogen receptor modulator, possessing agonist or antagonistic activity in different tissues. Thus, the objective of this study was to investigate the effect of TMX on the zymosan-induced arthritis model. Female Swiss normal and ovariectomized (OVX) mice were divided into groups and treated for five days with TMX (0.3, 0.9 or 2.7 mg/kg) or 17-β-estradiol (E2, 50 µg/kg). On the fifth day, arthritis was induced and 4 h later, leukocyte migration into joint cavities was evaluated. The neutrophil migration in OVX animals, but not in normal mice, treated with TMX (all tested doses) was significantly decreased compared with mice that received the vehicle (P≤0.05). Similarly, this effect was also demonstrated in the E2-treated group. Therefore, the present study demonstrates that TMX presented agonist effects in inhibiting neutrophil migration and preventing arthritis progression in OVX mice.

Além do impacto ambiental, a deriva de glyphosate pode causar danos sobre culturas sensíveis a esse herbicida. O objetivo deste estudo foi avaliar a influência de adjuvantes e pontas de pulverização na deriva de glyphosate aplicado via terrestre. O estudo foi conduzido em campo, arranjado em esquema fatorial (3 x 9), sendo o primeiro fator constituído por três pontas de pulverização: uma de jato plano (Teejet XR 110.015) e duas de jato plano com indução de ar (Teejet AIXR 110.015 e Teejet TTI 110.015). O segundo fator foi composto por sete adjuvantes: Break Thru(r) (0,1% v.v.), Grap' Oil(r) (0,5% v.v.), Grap Super Gun(r) (0,03% v.v.), Iharol(r) (1% v.v.), NP-10(r) (0,075 L ha-1), Dash HC(r) (0,5% v.v.) e Emultec R(r) (0,05 L ha-1). Os adjuvantes foram adicionados à calda de aplicação juntamente com o herbicida Roundup Transorb R(r) na dose de 2,16 kg e.a. ha-1. Além desses, foram incluídos dois tratamentos testemunha: um sem aplicação e outro com aplicação de glyphosate sem adjuvante. As variáveis avaliadas foram a distância percorrida pelas gotas para fora do alvo, a fitotoxicidade causada pela deriva, o diâmetro mediano volumétrico, a densidade de gotas e o volume recuperado pela utilização de papéis hidrossensíveis. As pontas de pulverização TTI 110.015 e AIXR 110.015 proporcionaram menor deriva que a ponta XR 110.015. As pontas TTI e AIXR apresentaram menor densidade de gotas e maior diâmetro mediano volumétrico, quando comparadas à ponta XR. A utilização dos adjuvantes NP-10(r) e Emultec R(r) com a ponta de pulverização XR 110.015 aumentou a deriva quando comparada com a aplicação de glyphosate sem adjuvante, porém não diferiram dos demais adjuvantes. O adjuvante Emultec R(r) aumentou o DMV das gotas, quando geradas pela ponta XR 110.015. A utilização dos adjuvantes Grap' Oil(r), Grap Super Gun(r), Iharol(r) e Dash HC(r) diminuiu a densidade de gotas derivadas, quando geradas pela ponta XR 110.015.

Glyphosate drift can cause environmental impact and also affect crops that are sensitive to this herbicide. The aim of this study was to evaluate the influence of different adjuvants and different nozzles on glyphosate drift in terrestrial applications. The study was conducted in the field, arranged in a factorial design (3 x 9): factor A consisted of three different spray nozzles: one flat spray nozzle (Teejet = XR 110.015) and two air induction flat spray nozzles (Teejet AIXR 110.015 and Teejet TTI 110.015). Factor B was composed of seven adjuvants: Break Thru(r) (0.1% v.v.), Grap' Oil(r) (0.5% v.v.), Grap Super Gun(r) (0.03% v.v.), Iharol(r) (1% v.v.), NP-10(r) (0.075 L ha-1), Dash HC(r) (0.5% v.v.) and Emultec R(r) (0.05 L ha-1). The adjuvants were added to Roundup Transorb R(r) herbicide at 2.16 a.i. ha-1. Moreover, there were two control treatments, one untreated check and another one with application of glyphosate without adjuvant. The distance of droplets off-target, injury caused by drift, volumetric median diameter (VMD), droplet density and volume were evaluated by water sensitive paper cards. The TTI 110.015 spray nozzle resulted in lesser drift when compared to the AIXR 110.015 and XR 110.015 nozzles. The TTI and AIXR nozzles showed lower droplet density and greater VMD compared with the XR nozzle. The use of NP-10(r) and Emultec R(r) adjuvants with spray nozzle XR 110.015 increases drift. Adjuvant Emultec R(r) increases VMD droplets when used with the XR 110.015 nozzle. The use of adjuvants Grap' Oil(r), Grap Super Gun(r), Iharol(r) and Dash HC(r) decrease density derived droplets when generated with the XR 110.015 nozzle.

A agenesia renal é uma afecção congênita rara na espécie felina, frequentemente associada a uma malformação reprodutiva. O presente trabalho relata o caso de um felino com agenesia renal unilateral associada a criptorquidismo ipsilateral, com ênfase no diagnóstico, tratamento e acompanhamento. O paciente foi conduzido ao Hospital de Clínicas Veterinárias da Universidade Federal do Rio Grande do Sul para avaliação de criptorquidismo. A agenesia renal foi um achado durante a ecografia abdominal do felino. Durante a laparotomia, foi confirmada a ausência do rim e ureter direito, hipertrofia do rim esquerdo e presença de um testículo ectópico. O paciente teve alta após a recuperação anestésica e se mantém clinicamente estável, transcorridos seis meses da cirurgia. A agenesia renal unilateral é uma condição compatível com a vida, contanto que o rim existente apresente funcionamento aceitável. Assim, sugere-se que a possibilidade de rim único em felinos criptorquidas deve ser investigada sempre que possível, tendo em vista a alta correlação entre essas malformações, e objetivando um acompanhamento da função renal do paciente ao longo da vida.

Renal agenesis is a rare disorder in feline species, commonly associated with reproductive malformation. This study aims to report the case of a cat with unilateral renal agenesis combined with ipsilateral cryptorchidism, emphasizing the diagnosis, treatment and patient follow up. The patient was taken to the Veterinary Hospital of the Federal University of Rio Grande do Sul to evaluate the cryptorchidism. The renal agenesis was an incidental finding during the abdominal ultrasound. At laparotomy, the absence of the right kidney and ureter was confirmed, hypertrophy of the left kidney and the presence of an ectopic testicle were found. The patient was discharged after recovering from anesthesia and remains clinically stable six months after surgery. The unilateral renal agenesis is a life compatible condition as long as the existing kidney has an acceptable performance. Therefore, it is suggested that the chance of a single kidney in cats whit cryptorchidism should be investigated, given the correlation between these malformations, and aiming to monitor renal function throughout the life of the patient.

A planta Sideroxylon obtusifolium, conhecida popularmente como "quixabeira", é utilizada na medicina popular para o tratamento de processos inflamatórios e dolorosos. O objetivo deste trabalho foi avaliar a possível ação cicatrizante, bem como anti-inflamatória tópica, do extrato etanólico (EE) da entrecasca da S. obtusifolium no modelo de ferida e cicatrização cutânea. Para isso, ratos Wistar foram anestesiados, submetidos à incisão dorsal, e discos de esponja de álcool polivinílico foram inseridos subcutâneamente. Quarenta e oito horas após os animais foram eutanasiados avaliando-se medidas de tensão de ruptura nas cicatrizes da pele dorsal, a atividade de mieloperoxidase (MPO), e a contagem de células nos exsudatos das esponjas. A atividade sequestradora do radical 2,2-difenil-1-picrilhidrazil (DPPH) foi determinada para avaliar a capacidade antioxidante do EE. Observamos que a administração tópica, 2 vezes ao dia, do EE da S. obtusifolium (3-30 mg 0,4 mL-1 de creme Lanette(r)) não alterou a tensão de ruptura das cicatrizes. Entretanto, o EE da S. obtusifolium (30 mg 0,4 mL-1 de creme Lanette(r)) foi capaz de diminuir a atividade de MPO (p<0,01) e a contagem celular (p<0,05) no exsudato das esponjas, após 48 h da indução da ferida. O EE mostrou-se ativo como sequestrador do radical DPPH. Os valores de IC50 para o EE e ácido gálico foram semelhantes (1,04 ± 0,27 e 1,15 ± 0,02 μg mL-1 de DPPH, respectivamente). Concluímos que o EE da entrecasca da S. obtusifolium possui atividade anti-inflamatória através da administração tópica no modelo de cicatrização cutânea em ratos, além de seu potencial antioxidante.

The Sideroxylon obtusifolium is a medicinal plant known as “quixabeira” in Portuguese that has been used in folk medicine for the treatment of inflammatory and painful processes. The aim of this study was to evaluate the effect of the ethanol extract (EE) of S. obtusifolium on the cicatrization and local anti-inflammatory activities using a model of wound healing in rats. Wistar rats were submitted to a dorsal skin incision and polyvinyl alcohol sponge discs were implanted. Animals were euthanized after 48 h, and the tensile strength measurement at the wound site, the myeloperoxidase (MPO) activity and the leukocyte counts in the wound exudates were evaluated. The 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity was determined to evaluate the antioxidant capacity of the EE. We observed that the topical administration of the EE of S. obtusifolium (3-30 mg 0.4 mL-1 of Lanette® cream) twice a day did not alter the tensile strength, but it decreased the MPO activity (p<0,01)and leukocyte infiltration (p<0,05) after 48 h of induction of wound. The EE shows to be active as DPPH radical scavenger. The IC50 value for the EE was similar to the IC50 value of gallic acid (1.04 ± 0.27 and 1.15 ± 0.02 μg mL-1 DPPH, respectively). In conclusion, the EE of Sideroxylon obtusifolium shows local anti-inflammatory activity in the model of wound healing in rats, in addition to antioxidant potential.

The objective of this study was to evaluate gastric emptying (GE) in pediatric patients with functional constipation. GE delay has been reported in adults with functional constipation. Gastric emptying studies were performed in 22 children with chronic constipation, fecal retention and fecal incontinence, while presenting fecal retention and after resuming regular bowel movements. Patients (18 boys, median age: 10 years; range: 7.2 to 12.7 years) were evaluated in a tertiary pediatric gastroenterology clinic. Gastric half-emptying time of water (reference range: 12 ± 3 min) was measured using a radionuclide technique immediately after first patient evaluation, when they presented fecal impaction (GE1), and when they achieved regular bowel movements (GE2), 12 ± 5 weeks after GE1. At study admission, 21 patients had reported dyspeptic symptoms, which were completely relieved after resuming regular bowel movements. Medians (and interquartile ranges) for GE1 and GE2 were not significantly different [27.0 (16) and 27.5 (21) min, respectively (P = 0.10)]. Delayed GE seems to be a common feature among children with chronic constipation and fecal retention. Resuming satisfactory bowel function and improvement in dyspeptic symptoms did not result in normalization of GE data.

A mancha bacteriana, causada por Xanthomonas axonopodis pv. passiflorae, é uma das mais importantes doenças do maracujazeiro, podendo limitar a produção dessa frutífera em algumas regiões do País. O uso de resistência genética e controle químico, juntamente com o emprego de medidas de exclusão, são as práticas de controle da doença mais recomendadas. Para o desenvolvimento de variedades resistentes é necessário conhecer tanto a variabilidade genética do hospedeiro quanto do patógeno. Nesse trabalho foi estudada a variabilidade de cinqüenta isolados patogênicos de Xanthomonas axonopodis pv. passiflorae, coletados em quatro diferentes locais no estado de São Paulo. No estudo da variabilidade genética foram usados dados de marcadores moleculares RAPD (Random Amplified Polymorphic DNA), os quais foram usados para o cálculo do coeficiente de similaridade de Dice entre os isolados, análise de variância molecular (AMOVA) entre e dentro das populações e agrupamento dos isolados pelo método UPGMA. Para análise da agressividade foram usados cinco isolados, mais divergentes, baseado no dendrograma. O coeficiente de similaridade variou entre 0,6887 e 0,9688. Na análise de agrupamento, os isolados foram separados em sete grupos e não houve relação evidente entre local de coleta com a composição dos grupos. Na análise da variância molecular (AMOVA) verificou-se que a maior parte da variabilidade genética está dentro das populações (89,4%) e apenas 10,6%, entre populações. Os resultados da análise de agrupamento e da AMOVA indicam que existe grande fluxo gênico entre isolados bacterianos nas regiões analisadas. No teste de patogenicidade verificou-se diferença significativa de agressividade entre os isolados. Os resultados demonstram a importância do conhecimento da variabilidade genética e da agressividade na seleção dos isolados para serem utilizados em testes de resistência genética no desenvolvimento de variedades resistentes.

The bacterial spot, caused by Xanthomonas axonopodis pv. passiflorae , is one of the most important diseases of passion fruit, which can drastically limit the production in some regions of Brazil. The use of genetic resistance, chemical control and the adoption of exclusion measures are the more recommended practices for the control of this disease. To produce the resistant varieties knowledge about the variability of both host and pathogen is needed. In this study, we investigated the variability of 50 pathogenic isolates of X. axonopodis pv. passiflorae, collected from four production areas in São Paulo State. We used RAPD markers (Random Amplified Polymorphic DNA) for identification of the degree of genetic similarities through dice coefficient, analysis of molecular variance (AMOVA), among as well as within populations and clustering by the UPGMA method. Based on the clustering, the five more divergent isolates were selected for agressiveness trials. The similarity coefficient ranged from 0.6887 to 0.9688. Clustering analysis grouped the isolates in seven categories and neither apparent correlation was identified among their geographical origin no clustering pattern. The AMOVA data showed that the variability within and among populations were 89.4% and 10.6% respectively. The clustering analysis indicated genic flow among the different production areas. The results highlights the importance of taking into consideration the bacterial genetic variability of this bacterium as well as its agressiveness during the selection of resistant varieties.

A diversidade morfológica da superfície foliar existente entre as espécies de plantas e a presença de estruturas foliares como tricomas, estômatos, cutícula e ceras podem exercer grande influência na aderência e deposição das gotas de pulverização, assim como na absorção do herbicida. Desta forma, o objetivo do presente trabalho foi estudar, em quatro espécies de plantas daninhas aquáticas (Enhydra anagallis, Eichhornia crassipes, Heteranthera reniformis e Typha subulata), a área de molhamento de gotas de pulverização nas superfícies foliares adaxial e abaxial, bem como o pH foliar. O experimento foi realizado em Botucatu-SP. As plantas foram cultivadas em caixas d'água sob condições de campo, e quando atingiram seu pleno desenvolvimento (antes do florescimento), foram realizadas as avaliações de pH foliar e da área de molhamento de gotas de pulverização. As tensões superficiais das gotas aplicadas (0,5 mL), apresentadas pelas soluções de glyphosate aplicado isolado a 5,0% v v-1 (Rodeo 480 g L-1 e.a. produto comercial), glyphosate + Aterbane BR (5,0% + 0,5% v v-1), glyphosate + Silwet L-77 (5,0% + 0,05% v v-1), além das soluções com os adjuvantes isolados, Aterbane BR (0,5% v v-1) e Silwet L-77 (0,05% v v-1) foram respectivamente 72,1; 28,7; 23,3; 37,3 e 22,1 mN m-1. As médias obtidas de pH foliar variaram entre 5,50 e 7,50, destacando-se a espécie E. anagallis com valores de 6,68 e 7,02 para as faces adaxial e abaxial, respectivamente. Dentre as plantas daninhas aquáticas avaliadas T. subulata foi a espécie que apresentou as maiores médias de área de molhamento nas faces adaxial e abaxial da folha, proporcionada pelas as soluções de glyphosate + Aterbane BR, glyphosate + Silwet L-77 e Silwet L-77, com valores de 12,99-7,03; 20,04-17,95 e 31,81-25,91 mm², respectivamente.

The morphological diversity of leaf surface and structures such as trichomes, stomata, cuticle, and waxes that exists among plant species can have great influence on the adherence and deposition of spray droplets, as well as on herbicide absorption. The aim of this research was to study leaf pH and to evaluate wetting areas after applications of solution surfactants on the following aquatic weeds: Enhydra anagallis, Eichhornia crassipes, Heteranthera reniformis, and Typha subulata. The aquatic weeds were grown in reservoirs containing water under open air conditions and their leaf tissues were collected when the plants reached full development (before flowering). The mean leaf pH varied between 5.50 and 7.50; E. anagallis should be pointed out for presenting the highest pH values, of 6.68 and 7.02 on the upper and lower leaf surfaces, respectively. Surface tension reduction for glyphosate alone (5.0% v v-1), glyphosate + Aterbane BR (5.0% + 0.5% v v-1), glyphosate + Silwet L-77 (5.0% + 0.05% v v-1), and surfactants alone, Aterbane BR (0.5% v v-1), and Silwet L-77 (0.05% v v-1), were, respectively: 72.1; 28.7; 23.3; 37.3, and 22.1 mN m-1.T. subulata was the aquatic species with the highest upper and lower leaf surface wetting area.

In this study, we identified disease resistance gene homologs in Brassica oleracea and assessed their expression in lines resistant and susceptible to Xanthomonas campestris pv. campestris (Xcc). Two DNA fragments of approximately 2.5 kb (BI-16/RPS2 and Lc201/RPS2) were amplified by PCR from two Brassica lines using primers based on an RPS2 homologous sequence previously described in the Brassica oleracea ecotype B117. The sequences of these fragments shared high similarity (95-98%) with RPS2 homologs from various Brassica species. The digestion of these fragments with restriction enzymes revealed polymorphisms at the Xba I restriction sites. The length polymorphisms were used as a co-dominant marker in an F2 population developed to segregate for resistance to Xcc, the causal agent of black rot. Linkage analysis showed no significant association between the marker and quantitative trait loci for black rot. RT-PCR with specific primers yielded an expected 453 bp fragment that corresponded to the RPS2 homologs in both resistant and susceptible lines inoculated with the pathogen, as well as in non-inoculated control plants. These results suggest that these homologs are constitutively expressed in B. oleracea.

A preservação adequada do DNA de um determinado organismo é fundamental para o sucesso no uso de técnicas moleculares como RAPD-PCR. O objetivo deste trabalho foi comparar métodos simples de preservação de espécimes de Dalbulus maidis (DeLong & Wolcott) (Hemiptera: Cicadellidae) quanto à quantidade e qualidade do DNA para uso em RAPD-PCR, após períodos sucessivos de armazenamento. Avaliaram-se oito métodos: congelamento (-20ºC); álcool etílico 70% (-20ºC e temperatura ambiente); álcool etílico absoluto (-20ºC e temperatura ambiente); secagem ao ar; e preservação em tampão de extração (inseto inteiro e macerado). A intervalos de 10-30 dias, o DNA foi extraído, quantificado e amplificado via RAPD-PCR pelo oligonucleotídeo OPA-04. A qualidade do DNA extraído foi observada em gel de agarose 0,8%. Após 210 dias de armazenamento, o congelamento (-20ºC) mostrou ser a melhor técnica. Quantidades satisfatórias de DNA também foram obtidas dos insetos conservados em álcool etílico absoluto (-20ºC), álcool etílico 70% (-20ºC) e tampão de extração (inseto inteiro e macerado). Insetos conservados em álcool etílico absoluto (temperatura ambiente), em álcool etílico 70% (temperatura ambiente) e secos ao ar mostraram-se impróprios para estudos com RAPD-PCR após 120, 60 e 10 dias de armazenamento, respectivamente.

The appropriate preservation of the DNA of a certain organism is important for successful application of molecular techniques like RAPD-PCR. This study was designed to compare simple methods of preservation of specimens of Dalbulus maidis (DeLong & Wolcott) (Hemiptera: Cicadellidae) with respect to quantity and quality of DNA for use in RAPD-PCR, after different storage periods. Eight methods were tested: freezing (-20ºC); ethyl alcohol 70% (-20ºC and room temperature); absolute ethyl alcohol (-20ºC and room temperature); air-dry; and preservation in extraction buffer (whole and homogenized insect). At intervals of 10 to 30 days, insect DNA was extracted, quantified and amplified through RAPD-PCR using primer OPA-04. The quality of extracted DNA was observed on 0.8% agarose gel. After 210 days of preservation, freezing (-20ºC) showed to be the best method. Satisfactory quantities of DNA were also obtained from insects conserved in absolute ethyl alcohol (-20ºC), ethyl alcohol 70% (-20ºC) and extraction buffer (whole and homogenized insect). Insects conserved in absolute ethyl alcohol (room temperature), ethyl alcohol 70% (room temperature) and air-dry conditions were inappropriate for RAPD-PCR studies after 120, 60 and 10 days of storage, respectively.

Em experimento realizado em casa de vegetação com quatro linhagens e dois híbridos de milho (Zea mays), verificou-se variabilidade para resistência a queima foliar causada por Colletotrichum graminicola avaliada por meio das variáveis área total de lesão por planta (ATL), área média de lesão (AML), comprimento total de lesão por planta (CTL), comprimento médio de lesão (CML) e número de lesões por planta (NL). Quando dois isolados de C. graminicola foram inoculados nessas linhagens e em híbridos de milho não foi verificada interação diferencial entre isolados e genótipos do hospedeiro. Em estudos de herança da resistência, as linhagens genitoras L186 e L64 comportaram-se como suscetível e resistente, respectivamente, ao passo que o híbrido L186 x L64 apresentou resistência. A análise das freqüências observadas de plantas resistentes e suscetíveis na população F2 resultantes da autofecundação do híbrido, indicou ocorrência de herança monogênica com dominância completa para o alelo que confere resistência à doença.

Variability in resistance to corn (Zea mays) leaf anthracnose caused by Colletotrichum graminicola was detected among four lines and two hybrids for the variables: total area of lesion (ATL), average lesion area (AML), total lesion length (CTL), average lesion length (CML) and lesion number (NL). No differential interaction was detected among isolates of C. graminicola and inbred lines and hybrids. In studies about the mode of resistance to foliar anthracnose, the inbred lines L186 and L64 were susceptible and resistant, respectively, whereas the hybrid L184 x L64 were resistant. The observed frequency of susceptible and resistant F2 plants derived from selfing of F1 plants from the cross L186 x L64 indicated monogenic and dominant control of resistance

Locos de microssatélites são úteis como marcadores genéticos pois são numerosos, ocorrem em todos os cromossomos e possuem um alto nível de polimorfismo. Além disso, podem ser amplificados por meio da PCR e resolvidos em géis de agarose. O maior problema, no entanto, reside no fato de que os primers utilizados em sua amplificação apresentam diferentes comprimentos e composição de nucleotídeos, o que gera a necessidade de otimizar programas de PCR para cada loco. Este trabalho descreve programas de PCR "touchdown" utilizados com sucesso na amplificação de um conjunto de 125 locos de microssatélites de milho escolhidos por serem representativos de todos os cromossomos. Os programas variaram tanto em relação à temperatura de anelamento (Tm), como na concentração dos primers. Assim, os primers puderam ser agrupados em quatro grupos: um grupo maior, constituído por primers que mostraram boa amplificação quando utilizados em um programa básico já publicado porém com concentrações diferentes de primers; um segundo grupo que amplificou em programas alternativos, nos quais as Tm foram modificadas de modo a incluir as Tm de ambos os primers ou a maior Tm do par de primers, estimadas em base da composição nucleotídica dos primers; um terceiro grupo foi composto por primers que amplificaram em programas com Tm mais elevadas que as estimadas, e um quarto grupo, com apenas dois pares de primers, em que a situação oposta ocorreu.

Maize (Zea mays L.) microsatellite loci are useful as genetic markers because they are numerous, occur in every chromosome, and have a high content of polymorphism information. Furthermore, they can be amplified by PCR and the resulting fragments resolved on agarose gels. A major problem, however, is that the primers used in their amplification often have different length and nucleic acid content, requiring optimization of PCR amplification programs for each locus. We developed "touchdown" PCR programs successfully used in the amplification of a set of 125-maize microsatellite loci, chosen as representative of all chromosomes. The programs varied both in relation to the annealing temperature (Tm) and primer concentration. Primers could be divided into four groups. A large group included primers that amplified well in a basic previously published amplification program but with different primer concentrations. A second group amplified in alternative programs in which the annealing temperatures were changed so as to include the Tm of either both primers or the highest Tm of the primer pair estimated based on their nucleotide composition. A third group included primers that amplified in programs with Tm higher than those estimated, and in a fourth group, with just two pairs of primers, the opposite situation prevailed.