Tissue culture parameters in sweet orange cultivars

The objective of this work was to establish tissue culture parameters for gene transfer in sweet orange cultivars. Epicotyl explants with different ages were cultured with 6-benzylaminopurine (BAP), kanamycin and hygromycin. Shoots were cultured with alpha-naphthaleneacetic acid (NAA) alone or in combination with indole-3-butyric acid (IBA). The requirement of BAP for shoot development was genotype-specific. Epicotyl explants from 35-day-old seedlings produced significantly more shoots per explant in 'Pêra'. Kanamycin inhibited shoot regeneration for the most cultivars. The percentage of shoots that produced roots in 'Pêra' was significantly higher in medium with NAA and IBA than with NAA alone.

Cell suspension culture and plant regeneration of a Brazilian plantain, cultivar Terra

The objective of this study was to establish cell suspension culture and plant regeneration via somatic embryogenesis of a Brazilian plantain, cultivar Terra Maranhão, AAB. Immature male flowers were used as explant source for generating highly embryogenic cultures 45 days after inoculation, which were used for establishment of cell suspension culture and multiplication of secondary somatic embryos. Five semisolid culture media were tested for differentiation, maturation, somatic embryos germination and for plant regeneration. An average of 558 plants per one milliliter of 5% SCV (settled cell volume) were regenerated in the MS medium, with 11.4 µM indolacetic acid and 2.2 µM 6-benzylaminopurine. Regenerated plants showed a normal development, and no visible somaclonal variation was observed in vitro. It is possible to regenerate plants from cell suspensions of plantain banana cultivar Terra using MS medium supplemented with 11.4 µM of IAA and 2.2 µM of BAP.

Growth regulators, culture media and antibiotics in the in vitro shoot regeneration from mature tissue of citrus cultivars

The objective of this study was to evaluate the effects of 6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA) combinations, basal media and beta-lactam antibiotics on in vitro organogenesis from mature stem segments of 'Pêra', 'Valência' and 'Bahia' sweet oranges and 'Cravo' rangpur lime. For induction of shoot regeneration, the segments of the four cultivars were placed on Murashige and Skoog (MS) medium containing the following BAP/NAA concentrations: 0.0/0.0; 0.25/0.0; 0.25/0.25; 0.5/0.0; 0.5/0.5; 1.0/0.0; 2.0/0.0; 2.0/0.25; 2.0/0.5; and 2.0/1.0 mg L-1. In order to test the influence of the culture media on shoot-bud induction, (MS), Murashige and Tucker (MT), and woody plant medium (WPM) formulations were evaluated, associated with the best combination of plant growth regulators obtained in the previous experiment. The influence of four beta-lactam antibiotics (timentin, cefotaxime sodium salt, meropenem trihydrate and augmentin) on shoot regeneration was determined. Better regeneration responses were achieved when internodal segments were cultured onto MS-based medium with 500 mg L-1 cefotaxime with the following BAP/NAA concentrations: 0.5 + 0.25 mg L-1 for 'Cravo', 1.0 + 0.25 mg L-1 for 'Valência' and 'Bahia', and 1.0 + 0.5 mg L-1 for 'Pêra'. Genotype, growth regulators, basal media and beta-lactam antibiotics affect the morphogenetic response in mature tissues of citrus.

In vitro organogenesis from internodal segments of adult sweet orange plants

The objective of this work was to optimize in vitro plant regeneration via organogenesis from tissues of adult 'Hamlin', 'Pêra', and 'Valência' sweet orange plants. Explants were grown in EME culture medium with different concentrations of 6-benzylaminopurine (BAP) and naphthaleneacetic acid (NAA), at 27ºC in the absence of light for 50 days, followed by a 16-hour photoperiod for 20 days. Regeneration was assessed 50 and 70 days after in vitro culture. Organogenesis in cultivars Hamlin and Valência was promoted by EME supplemented with BAP, while NAA showed no apparent effect.

In vitro organogenesis in some citrus species

In vitro organogenesis of Citrus was studied for the genotypes Citrus sinensis cv. 'Natal', C. limonia, C. volkameriana, and C. aurantium, with the use of epicotyl segments-derived explants, cultured in MT salts and vitamins medium supplemented with different concentrations of 6-benzylaminopurine (BAP - 0.0; 0.5; 1.0; 1.5 or 2.0 mg L-1). For the recalcitrant genotypes C. limonia and C. aurantium the in vitro organogenesis was also studied with internodal segments-derived explants, cultured in MT salts and vitamins medium supplemented with 0; 0.5; 1.0; 2.0, or 4.0 mg L-1 of BAP. The efficiency of culture medium supplementation with the combination of BAP (0.0; 1.0, or 2.0 mg L-1) and NAA (1-naphthaleneacetic acid - 0.0; 0.3, or 0.5 mg L-1) in the development of adventitious shoots was evaluated for C. aurantium. Culture medium supplementation with BAP is not essential for the adventitious shoots development in the four genotypes studied when epicotyl segments-derived explants are used. In general, culture media supplementation with BAP decreased the percentage of responsive explants excepted for C. sinensis cv. 'Natal' and C. limonia when the concentrations of 1.5 and 2.0 mg/L were used. The presence of cytokinin, in concentrations up to 2 mg/L, stimulated the in vitro organogenesis when internodal segments-derived explants were used for C. limonia and C. aurantium. For C. aurantium no adventitious shoots developed in explants (internodal segments) cultured in basal culture medium, without BAP supplementation. Although no statistic differences could be detected, culture media supplementation with the combination of BAP and NAA favored the development of adventitious shoots in C. aurantium. The best concentration of NAA varied according to BAP concentration. The results presented herein, show that Citrus in vitro organogenesis depends on the interaction of culture medium composition, explant differentiation level, and genotype.

Leaf and root volatiles produced by tissue cultures of Alpinia zerumbet (pers.) Burtt & Smith under the influence of different plant growth regulators

Volatiles produced by plantlets of Alpinia zerumbet were obtained by means of simultaneous distillation-extraction (SDE). The effects of indole-3-acetic acid, kinetin, thidiazuron and 6-benzylaminopurine on leaf and root volatile composition obtained by tissue cultures were investigated. A higher content of b-pinene and a lower content of sabinene were observed in leaf volatile of plantlets cultured in control, IAA and IAA+ TDZ media, as compared with those of donor plants. In vitro conditions were favorable to increase caryophyllene content. Volatile compounds from the root were characterized mainly by camphene, fenchyl-acetate and bornyl acetate; which constitute about 60% of total volatile.

MICROPROPAGATION AND ACCLIMATIZATION OF Aegiphila verticillata Vell.: AN ENDANGERED WOODY SPECIES

The objective of this work was to establish an efficient protocol for in vitro multiplication and rooting, as well as ex vitroacclimatization of Aegiphila verticillata, a woody species found in Brazilian rocky fields. Aseptic cultures were established by seeds and two multiplication analyses were performed. In the first, we employed 6-benzylaminopurine (BAP – 0, 2.5, 5 and 7.5 μM) + α-naphthalene acetic acid (NAA – 0, 0.2, 0.4 and 0.6 μM) and, in the second, were studied adenine sulfate, kinetin and thidiazuron (0, 5, 7.5, 10 and 12.5 μM). After 90 days, we assessed the quantitative and qualitative shoot propagation. There were more than 90% seed germination and low contamination (2%). In multiplication phase, the culture medium that promoted the best quantitative and qualitative culture development was supplemented with 7.5 μM BAP + 0.4 μM NAA. In the rooting assay, were used NAA, indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) (0, 0.1, 0.2, 0.3 or 0.4 μM). After 90 days, the root number and rooting quality were evaluated. In this analysis, differences were not found between the control and the other treatments. Rooted plantlets were acclimatized in styrofoam trays for 30 days, after which they were transferred to pots in the greenhouse. Only 3% of the plants subjected to initial acclimatization died and 70% of the plants transferred to the field conditions survived and showed normal development. The results founded in this work are the first involving in vitro propagation and ex vitroacclimatization of Aegiphila verticillata and provide a continuous supply of this medicinal native species, endangered due anthropogenic activities.

Sinningia allagophylla(Gesneriaceae): in vitro cultivation of a native plant of the Brazilian cerrado

We used axillary buds as initial explants for hormone interaction studies required for in vitro cultivation of S. allagophylla. Callus production was achieved on gelled Murashige & Skoog medium (MS) supplemented with indole-3-acetic acid (IAA= 0.1 and 0.5 mg.l­1 alone or combined with 6 benzylaminopurine) (BA= 0.01 and 0.1 mg.l-1). A hormone balance between IAA and BA that would encourage shoot bud development was not found. Nodal segments from axenic cultures grown in the presence of cytokinin (0.1 mg.1­1 of BA) without any auxin on MS medium with half-strength macronutrients were used as a standard explant source for subsequent experiments on optimum mineral culture media composition for S. allagophylla in vitro cultivation. We found that explants kept in vitro on gelled Gamborg et al. (B5) mineral composition culture medium showed better shoot and specially root growth than on MS medium. Comparisons of the ammonium and nitrate ratios of MS and B5 media indicate that B5 medium has a substantial reduced ammonium ion when compared to MS medium, as well as a lower total nitrogen level. The growth response pattern obtained in vitro may be evidence of the adaptation of this species to soils of poor mineral composition as found in the Brazilian cerrado, as well as an indication that nitrogen levels play a key role for S. allagophylla growth.

Papilomatose laríngea: análise morfológica pela microscopia de luz e eletrônica do HPV-6

Papilomatose laríngea é neoplasia benigna mais freqüente nas crianças, causada pelo HPV, principalmente subtipos 6 e 11 e caracteriza-se pela presença de lesões proliferativas exofíticas e recidivantes sobre a mucosa das vias aérea, em especial na laringe. Forma de Estudo: Clínico prospectivo. OBJETIVOS: Demonstrar alterações epiteliais morfológicas (pela microscopia de luz e eletrônica) em lesões papilíferas casadas pelo HPV-6. MATERIAL E MÉTODOS: Fragmentos de lesões de papilomatose laríngea, colhidos durante procedimento cirúrgico de quatro crianças (1 masculino, 3 femininas), foram submetidos à tipagem do HPV (por método de PCR), análise pela microscopia de luz e microscopia eletrônica (varredura e transmissão). RESULTADOS: Na tipagem, todos os papilomas eram do subtipo 6. A microscopia de varredura identificou projeções epiteliais de vários tamanhos, com células superficiais em descamação. A microscopia de luz demonstrou lesões exofíticas, revestidas por epitélio hiperplásico com coilócitos e binucleações, característicos do HPV. A membrana basal e o córion adjacente estavam íntegros. À microscopia eletrônica de transmissão identificou-se vacuolização perinuclear e alargamento das junções intercelulares. CONCLUSÕES: As alterações morfológicas apresentadas pelo HPV-6 demonstram o caráter não-invasivo da lesão, sendo necessário estudos morfológicos adicionais relacionando os outros tipos de HPV, considerados mais agressivos, com os achados ultra-estruturais.

Emergência e desenvolvimento de mudas de tomate IPA 6 em substratos, contendo esterco ovino

A busca por substratos alternativos, que possam substituir produtos comerciais na produção de mudas de tomate, é de fundamental importância para a redução dos custos nessa fase inicial da cultura. Objetivou-se, com esta pesquisa, avaliar a qualidade de mudas de tomate industrial IPA 6, produzidas com substratos à base de esterco ovino. O experimento foi conduzido em viveiro da Universidade Federal Rural de Pernambuco, Unidade Acadêmica de Serra Talhada, de 16 de fevereiro a 30 de março de 2012. Utilizou-se o delineamento inteiramente casualizado, com quatro repetições. Os tratamentos avaliados foram: T1 - [substrato comercial]; T2 - [esterco ovino + areia (1:1)]; T3 - [esterco ovino + areia (2:1)]; T4 - [esterco ovino + areia (3:1)]; T5 - [esterco ovino + solo (1:1)]; T6 - [esterco ovino + solo (2:1)]; T7 - [esterco ovino + solo (3:1)]; T8 - [esterco ovino + areia + solo (1:1:1)];T9 - [esterco ovino + areia + solo (2:1:1)]; T10 - [esterco ovino + areia + solo (3:1:1)]. Os indicadores de emergência avaliados foram: percentagem, índice de velocidade e tempo médio de emergência. Analisaram-se as seguintes características de desenvolvimento: número de folhas, altura de plântula, diâmetro de caule, comprimento de raízes, massa da matéria seca da parte aérea e das raízes. O substrato comercial proporcionou melhor vigor de mudas de tomate IPA 6. Por outro lado, o uso do esterco ovino mais solo (2:1), para compor substratos alternativos constituiu-se opção viável para a produção de mudas desse cultivar.

Identificação do enfermeiro de saúde pública na força de trabalho de Enfermagem de Saúde Pública no Departamento Regional de Saúde-6 de Ribeirão Preto, SP (Brasil)

Objetivou-se identificar o enfermeiro na força de trabalho de Enfermagem em Saúde Pública, no Departamento Regional de Saúde - 6 de Ribeirão Preto da Secretaria de Estado da Saúde, destacando e analisando alguns aspectos que o caracterizam. Utilizou-se, como fonte primária e direta de coleta de dados, questionário contendo dados de identificação, nível de formação, progressão funcional, funções, expectativas futuras quanto ao exercício profissional e filiação a órgãos de classe. O total do grupo estudado foi de 35 indivíduos do sexo feminino, predominando: grupo etário de 20 a 30 anos (40,0%); casados (51,5%); tempo de serviço entre zero e 15 anos (77,2%). Com referência à sua interproporcionalidade com as demais categorias, as enfermeiras se distribuíam nas seguintes relações: 1 enfermeira/13,5 pessoal auxiliar e 1 enfermeira/9,1 médicos. Quanto à formação profissional, 71,5% realizaram habilitação em Enfermagem em Saúde Pública e 8,6% Especialização em Saúde Pública. Não havia para as enfermeiras carreira nem quadro e seus vencimentos estavam entre 2 e 7 salários mínimos e apenas uma com 10. A função administrativa é exercida por todas as enfermeiras com predomínio de freqüência entre as inspetoras e de CS, 68,5%. As enfermeiras sentem-se preparadas para participar dos programas de assistência primária e 84,3% delas pretendem permanecer em exercício até sua aposentadoria.

Studies on mosquitoes (Diptera: Culicidae) and anthropic environment: 6 - Breeding in empty conditions of rice fields in South-Eastern Brazil

Studies on culicid breeding in empty rice fields were carried out during the cultivation cycle from May to November 1993. This period corresponded to stages 1 and 2, when empty conditions prevailed. Breeding occurred in stage 1 and the first part of stage 2, corresponding respectively to fallow uncultivated and ploughing situations. No breeding was found to take place during the second part of stage 2 when transient floods and harrowing occurred. The predominant species were Aedes scapularis, Culex nigripalpus and Cx. mollis. The Pilosus Group of Culex (Melanoconion) was found at lower densities. Some epidemiological considerations are presented.

Prevalência de cárie dentária em crianças da faixa etária de 0 a 6 anos matriculadas em creches: importância de fatores socioeconômicos

Foi avaliada a prevalência de cárie na dentição decídua de crianças entre 0 e 6 anos, matriculadas em creches dos Municípios de Bauru e São Paulo, SP (Brasil). O primeiro grupo (Bauru) não recebia cuidados sistematizados de saúde na instituição e o segundo (São Paulo) apresentava uma rotina de cuidados como norma institucional. Foram analisadas as variáveis relativas aos modos de viver desses grupos populacionais e sua associação com a ocorrência de cárie, efetuando um estudo de caso para caracterização de fatores coletivos de risco à cárie. Através de análise de regressão múltipla, verificou-se a influência da idade e freqüência de consultas odontológicas sobre a prevalência de cárie na amostra estudada (p<0,05). Na faixa etária de 5-6 anos, 23,3% das crianças de Bauru e 9,3% de São Paulo estavam isentas de cárie, contra a expectativa de 50% prevista na Meta n.o 1 da Organização Mundial da Saúde para o ano 2000. A prevalência de cárie foi mais elevada em Bauru nas crianças de 3-4 e 5-6 anos, apresentando significância estatística apenas para o grupo 3-4 anos (p<0,05). Não foram observadas diferenças estatisticamente significantes entre os sexos quanto à ocorrência de cárie.

Prevalência de cárie e necessidades de tratamento em escolares de 6 a 12 anos da rede pública de ensino

OBJETIVO: Conhecer a prevalência de cárie e necessidades de tratamento em escolares do interior do Estado de Goiás, Brasil. MÉTODOS: A amostra foi constituída de 1.419 escolares de 6 a 12 anos de idade, de ambos os sexos, que freqüentavam 25 escolas públicas na zona urbana de 9 municípios. RESULTADOS: A percentagem de escolares livres de cárie foi muito baixa em todas as idades, sendo 4,4% aos 12 anos. O índice CPO-D variou de 0,41 aos 6 anos a 5,19 aos 12 anos. O índice ceo-d nesta faixa etária variou de 4,93 a 0,29. As necessidades de tratamento superaram as necessidades atendidas, tanto na dentição decídua quanto na permanente. CONCLUSÃO: A alta prevalência de cárie em escolares do interior de Goiás sugere a necessidade de se implantar medidas educativas e preventivas em saúde bucal que intervenham nos reais determinantes da doença na população.