Increased systemic IL-6 levels point to inflammation as a determinant of renal cell carcinoma development

Introduction: Renal cell carcinoma (RCC) is one of the most prevalent kidney tumors. Inflammation is believed to be a key factor in its progression and spread since inflammatory markers are generally associated with poor prognosis in RCC patients. Cytokines are cell communication molecules involved in both healthy and pathological processes, including tumor growth and progression. Recent findings suggest that cytokine level measurements could be used for cancer monitoring and prognosis. Methods: This study characterized and compared the levels of different cytokines associated with the classical Th1, Th2, and Th17 immune responses in plasma samples from RCC patients (n = 25) and healthy controls (n = 29). Cytokine levels (IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ, and IL-17A) were evaluated by flow cytometry using a BD Cytometric Bead Array (CBA) kit. Results: No statistical differences in systemic IL-2, IL-4, IL-10, IL-17A, TNF, and INF-γ levels were observed between RCC patients and controls (p > 0.05). However, higher systemic IL-6 levels were observed in RCC patients (p = 0.0034). Conclusions: This study highlights the importance of assessing the impact of IL-6 on RCC pathogenesis and its potential role as a biomarker of disease progression. (AU)

Inducción de tormenta de citocinas en pacientes infectados con SARS- CoV-2 y desarrollo de COVID-19: Tiene el tracto gastrointestinal alguna relación en la gravedad? / Induction of "Cytokine storm" in patients infected with SARS-CoV-2 and development of COVID-19. Does the gastrointestinal tract any relation in severity?

Los coronavirus son una extensa familia de virus; algunos pueden ser causa de diversas enfermedades humanas, que van desde el resfriado común hasta el síndrome respiratorio agudo severo. La enfermedad por coronavirus (COVID-19) es causada por el coronavirus zoonótico SARS-CoV-2, y comenzó en China en diciembre de 2019. En marzo de 2020, la Organización Mundial de la Salud (OMS) lo declaró una pandemia mundial. Esta enfermedad tiene un gran espectro de signos y síntomas, predominantemente se presenta con tos y fiebre, lo que resulta en una proporción de pacientes que desarrollan síndrome de dificultad respiratoria aguda (SDRA). Se ha planteado que la afectación pulmonar grave por el virus puede ser causada también por hiperinflamación y un cuadro similar a la linfohistiocitosis hemofagocítica (HLH), el cual es un síndrome clínico provocado por una respuesta inadecuada del sistema inmunológico a un desencadenante, sea infec-cioso, neoplásico, metabólico o reumatológico, que resulta en una reacción inflamatoria desproporcionada. Esta respuesta inmune altamente activada causa una liberación exagerada o tormenta de citocinas respon-sable del cuadro clínico. De allí la importancia de revisar el concepto actual y su desenlace en COVID-19. Recientemente se han publicado artículos en los que se sugiere que la presencia de síntomas gastrointesti-nales se ha asociado a COVID 19 con mayores indicadores de gravedad, lo que motiva al gastroenterólogo a explorar cómo el sistema digestivo podría impactar en esta hiperinflamación.(AU)

Coronaviruses are an extensive family of viruses, some of them can produce different kind of human diseases, ranging from the common cold to severe acute respiratory syndrome. Coronavirus disease (COVID-19) is caused by the SARS-CoV-2 zoonotic coronavirus and started in China in December 2019. In March 2020, the World Health Organization (WHO) declared it a global pandemic. This disease has a wide spectrum of signs and symptoms, predominantly presenting with cough and fever, resulting in a proportion of patients who develop acute respiratory distress syndrome (ARDS). It has been argued that severe lung involvement by the virus may also be caused by hyperinflammation and a condition similar to hemophagocytic lymphohistiocyto-sis (HLH), which is a clinical syndrome caused by an inadequate response of the immune system to a trigger, whether infectious, neoplastic, metabolic, or rheumatologic, leading to a disproportionate inflammatory reac-tion. This activated immune response producing an exaggerated release or storm of cytokines responsible for this disease. There are recently publications suggesting that the presence of gastrointestinal symptoms is associated with COVID 19 disease with higher indicators of severity, which motivates the gastroenterologist to explore how the digestive system could impact this hyperinflammation

Use of plant growth regulators in fig tree seedling s 'roxo de valinhos' / Uso de reguladores vegetais em mudas de figueira 'Roxo de Valinhos'

The application of plant growth regulators can promote better development of the seedlings, essential for the success of fruit growing. The current study aims to evaluate the use of plant growth regulators in fig tree seedlings 'Roxo de Valinhos'. The experiment was carried out in a suspended nursery covered with plastic, using rooted cuttings of the cultivar Roxo de Valinhos. Four applications were made with a commercial product, known as Stimulate®, at doses of 0, 50, 100, 150, and 200 mL L-1. Using a randomized block experimental design, the study was subdivided into plots (doses X days of the last application [after 0, 7, 14 and 21]). The following traits were evaluated: stem and root length; stem diameter; root volume; leaf number; leaf area; stem, leaf and root dry mass; specific leaf area, leaf weight ratio; leaf area ratio; and chlorophyll total. Results indicated that the product promoted greater seedlings development at a dose range from 100 to 150 mL L-1. Different dose levels did not vary according to the days after product application. Furthermore, using such technique enabled seedlings of higher quality and vigour.

A aplicação de reguladores vegetais pode configurar uma técnica viável para promoção do melhor desenvolvimento das mudas, imprescindível para o sucesso da fruticultura. Deste modo, objetivou-se avaliar o uso de reguladores vegetais na produção de mudas da figueira 'Roxo de Valinhos'. O experimento foi realizado em viveiro suspenso com cobertura plástica, utilizando-se estacas enraizadas da cultivar Roxo de Valinhos. Foram realizadas quatro aplicações com o produto comercial Stimulate® nas concentrações de 0, 50, 100, 150, e 200 mL L-1. Utilizou-se o delineamento experimental em blocos casualizados com parcelas subdivididas [concentrações X épocas de avaliação (após 0, 7, 14 e 21 dias da última aplicação)]. Avaliou-se: comprimento do ramo e da maior raiz; diâmetro do ramo; volume de raízes; número de folhas e área foliar; massa seca de folhas, ramos e raízes; área específica foliar, razão de peso foliar e razão da área foliar; e teores de clorofila total. O uso do produto, nas concentrações de 100 a 150 mL L-1, promoveu maior desenvolvimento das mudas. O efeito das diferentes doses não variou em função dos dias após aplicação do produto. O uso de tal técnica permite a obtenção de mudas de maior qualidade e vigor.

Falta de asociación entre inflamación subclínica evaluada por ultrasonido y niveles de citocinas en artritis reumatoide / Lack of association between ultrasound-defined subclinica inflammation and cytokines levels in rheumatoid arthritis

Antecedentes: La artritis reumatoide (AR) es una enfermedad autoinmune crónica que se caracteriza por proliferación sinovial, ruptura de cartílago y destrucción ósea. Los biomarcadores en AR no se utilizan en forma rutinaria para evaluar la inflamación y tampoco la remisión. El ultrasonido musculoesquelético (US) visualiza los cambios en las articulaciones y el daño morfoestructural, mejorando la evaluación de la sinovitis.Objetivo: Identificar y describir la inflamación subclínica en pacientes con AR en re-misión, utilizando US.Métodos: Se incluyeron pacientes con AR en remisión. Se realizó una evaluación clí-nica con DAS28; se tomó muestra de sangre para analizar citocinas. Un ecografista reumatólogo sin acceso a datos clínicos realizó un conteo ecográfico utilizando el sco-re-7. Se utilizaron parámetros de tendencia central, análisis de correlación bivariada y X cuadrado. Se estableció un nivel de confianza del 95% y, por tanto, cualquier valor p ≤0.05 se consideró significativo.Resultados: Se incluyeron 38 pacientes con AR. La edad media fue de 45,26±12,24 años. Los niveles de citocinas asociadas al tiempo de la AR desde la remisión, no fue-ron estadísticamente significativas. El ultrasonido en los pacientes evidenció al menos una de las lesiones elementales; en escala de grises, la sinovitis ocurrió en un 94,7%; sinovitis con señal Doppler de poder (DP) 52,6%; en cuanto a erosiones, se registra-ron, respectivamente, un 55,3% en escala de grises y un 15,8% con DP. DAS28 >2,04 fue positivo al asociarse con el recuento de articulaciones dolorosas y significativo (p=0,009). Conclusión: La asociación entre la sinovitis clínica y en ecografía no tiene correlación con los criterios de AR en remisión, independientemente de cuán estricta sea su aplicación.

Background: Rheumatoid arthritis (RA) is a chronic autoimmune disease, character-ized by synovial proliferation, cartilage breakdown and bone destruction. Biomarkers are not routinely used to evaluate inflammation neither remission. Musculoskeletal ultrasound visualizes joint changes and morpho-structural damage improving the as-sessment of synovitis.Objective: To identify and describe subclinical inflammation in patients with RA in remission using US.Methods: RA patients in remission were included. A clinical evaluation and DAS28 score performed; a blood sample took to analyze cytokines. A rheumatologist ultraso-nographer blinded to clinical data performed a US 7-score joint count. Central tenden-cy parameters, bivariate correlation analysis, and X Square were used. A confidence level of 95% was set and, therefore, any p-value ≤0.05 was considered as significant.Results: 38 RA patients were included. Mean age was 45.26±12.24 years. Cytokines associated with the time since remission was not statistically significant. Patients dis-played at least one of US elementary lesions; gray-scale synovitis occurred in 94.7%; synovitis with PD signal 52.6%; gray-scale erosions 55.3% and erosions with PD 15.8% respectively. DAS28 >2.04 positive for tender joint count was significant (p=0.009).Conclusion: The association between the clinical and US synovitis does not correlate with RA remission criteria no matter how strict is its application.

Plant growth regulators and shade on emergence in macadamia nut tree / Reguladores vegetais e sombreamento na emergência de plântulas de macadâmia

This study aimed to evaluate the effect of plant growth regulators and shading on seed germination of macadamia nut trees. The experiment was conducted in a seedling production nursery of the company "QueenNut Macadâmia" located in the "Palmeiras" Farm, Dois Córregos, São Paulo (SP) state, Brazil. The Treatments were T1: water (control) under a shading screen, T2: GA4+7 + N-(phenylmethyl)-aminopurine (Promalin®) at 200 mL L-1 under a shading screen, T3: Promalin® at 400 mL L-1 under a shading screen, T4: gibberellic acid (GA3) + kinetin (Kt) + 3- indolebutyric acid (IBA) ­ Stimulate® at 5 mL kg-1 seeds under a shading screen, T5: Stimulate® at 10 mL kg-1 of seeds under a shading screen, and T6: water (control) under no shading screen. Seeds were soaked in the solutions or in water containing plant growth regulators for 24 hours. Then, they were dried and sown in sand. 450 seeds were used for each treatment. The evaluations began from seedling emergence by counting the total number of seedlings per treatment. A high average of macadamia nut tree seedling emergence was obtained in the treatments GA4+7 + N-(phenylmethyl)-aminopurine (Promalin®) 400 mL L-1 with Sombrite® (75.7%), GA4+7 + N-(phenylmethyl)-aminopurine (Promalin®) 200 mL L-1 (72.6%) Sombrite® and water (control) without Sombrite® (71.5%).

Este estudo objetivou avaliar os efeitos de reguladores vegetais e sombreamento na emergência de plântulas de macadâmia. O experimento foi conduzido no viveiro de produção de mudas de macadâmia da empresa QueenNut Macadâmia, na Fazenda Palmeiras no município de Dois Córregos-SP. Os tratamentos utilizados foram: T1. água (testemunha) com Sombrite®, T2. GA4+7 + N-(fenilmetil)-aminopurina (Promalin®) a 200 mL L-1 com Sombrite®, T3. Promalin® a 400 mL L-1 com Sombrite®, T4. GA3 + IBA + Kt ­ Stimulate® a 5 mL kg-1 sementes com Sombrite®, T5. Stimulate® a 10 mL kg-1 sementes com Sombrite® e T6. água (testemunha) sem Sombrite®. As sementes foram embebidas em água ou nas soluções contendo os reguladores vegetais durante 24h, secas à sombra e semeadas em sementeiras de areia. Foram utilizadas 450 sementes em cada tratamento. A partir da emergência das plântulas foram iniciadas as avaliações, realizadas através da contagem do número total de plântulas emergidas por tratamento. Os melhores resultados na emergência de plântulas de macadâmia foram obtidos nos tratamentos com GA4+7 + N-(fenilmetil)- aminopurina (Promalin®) a 400 mL L-1 com Sombrite® (75,7%), GA4+7 + N-(fenilmetil)-aminopurina (Promalin®) a 200 mL L-1 (72,6%) com Sombrite® e água (testemunha) sem Sombrite® (71,5%).

In vitro germination, callus induction and phenolic compounds contents from Pyrostegia venusta (Ker Gawl) Miers / Germinação in vitro, indução de calos e teores de compostos fenólicos em Pyrostegia venusta (Ker Gawl) Miers

A protocol for the in vitro germination and callus induction in Pyrostegia venusta, a medicinal plant species from the Brazilian savannah (Cerrado) is provided. The establishment of cultures of P. venusta was originally obtained from seeds germinated in vitro and induced callus directly from leaf explants of seedlings. Seeds were germinated on MS and WPM media containing 100 and 50% of salt concentration, supplemented with 30 g L-1 sucrose. Callus induction consisted of the inoculation of leaf segments on MS medium plus 2,4-D or BAP in the presence or absence of light. The germination percentage averaged 85% and the aerial parts and roots of seedlings obtained in WPM with 50 and 100% of salt concentration showed elevated contents of total phenolic compounds and flavonoids compared to those obtained on MS medium. Calli induced with lower concentrations of 2,4-D had higher fresh and dry weight values. All treatments resulted in obtaining calli with contents of total phenolic compounds and flavonoids at or above the initial explant, highlighting treatments containing 9.05 µM 2,4-D and 8.88 µM BAP in the presence and absence of light, and 17.75 µM BAP in the absence of light.

É apresentado um protocolo para a germinação in vitro e indução de calos em Pyrostegia venusta, uma espécie medicinal do Cerrado. O estabelecimento de culturas foi obtido a partir de sementes germinadas in vitro e a indução de calos diretamente de explantes foliares das plântulas obtidas. Sementes foram germinadas em meios MS e WPM contendo 100 e 50% da concentração de sais, suplementado com 30 g L-1 de sacarose. A calogênese consistiu na inoculação de segmentos foliares em meio MS acrescido de 2,4-D ou BAP, na presença ou ausência de luz. A percentagem de germinação foi 85%, em média. Partes aéreas e raízes de plântulas obtidas em meio WPM com 50 e 100% da concentração de sais, apresentaram elevados teores de compostos fenólicos totais e flavonoides, em relação àquelas obtidas em meio MS. Calos induzidos com menores concentrações de 2,4-D apresentaram maiores valores de matéria fresca e seca. Todos os tratamentos proporcionaram a obtenção de calos com teores de compostos fenólicos totais e flavonoides iguais ou superiores ao explante inicial, com destaque para os tratamentos com 9,05 µM de 2,4-D e 8,88 µM de BAP, na presença e ausência de luz e 17,75 µM de BAP, na ausência de luz.

In vitro callus induction and estimation of plumbagin content from Plumbago auriculata Lam.

The medicinal plant Plumbago contains a very potent secondary metabolite, plumbagin having many therapeutic properties. Callus culture was induced using explants, leaf, stem and shoot apex, from P. auriculata. Murashige and Skoog media fortified with various growth hormones like NAA, IAA, IBA and 2, 4-D individually and in various combinations were checked for callus induction. Among the growth hormones used, 1 mg/L 2, 4-D showed best callusing. The hormonal combinations of 1 mg/L IAA and 1.5 mg/L NAA in the media exhibited best callus induction using stem internode as an explant. Plumbagin content from root, stem, leaf and callus was analyzed by using thin layer chromatographic technique. The callus derived from stem showed comparable plumbagin content to the in vivo plant parts. Quantitative spectrophotometric analysis of plumbagin from plant samples and callus indicated that plumbagin content was maximum in roots which was followed by callus, stem and leaf samples respectively. Generation of in vitro sources for plumbagin, for therapeutic applications will serve as a continuous supply and will contribute to preserve the natural plant recourses.

In vitro regeneration and ploidy level analysis of Eulophia ochreata Lindl.

Various parameters including explant-type, medium compositions, use of phytohormones and additives were optimized for direct and indirect regeneration of E. ochreata, a medicinal orchid under threat. Protocorm-like-bodies (PLBs) proved to be the best explants for shoot initiation, proliferation and callus induction. Murashige and Skoog’s (MS) medium containing 2.5 mg L-1 6-benzylaminopurine (BAP), 1.0 mg L-1 kinetin (Kin) and additives (adenine sulfate, arginine, citric acid, 30 mg L-1 each and 50 mg L-1 ascorbic acid) was optimal for shoot multiplication (12.1 shoots and 7.1 PLBs per explant with synchronized growth), which also produced callus. Shoot number was further increased with three successive subcultures on same media and ~40 shoots per explant were achieved after 3 cycles of 30 days each. Additives and casein hydrolysate (CH) showed advantageous effects on indirect shoot regeneration via protocorm-derived callus. Optimum indirect regeneration was achieved on MS containing additives, 500 mg L-1 CH, 2.5 mg L-1 BAP and 1.0 mg L-1 Kin with 30 PLBs and 6 shoots per callus mass (~5 mm size). The shoots were rooted (70% frequency) on one by fourth-MS medium containing 2.0 mg L-1 indole-3-butyric acid, 200 mg L-1 activated charcoal and additives. The rooted plantlets were hardened and transferred to greenhouse with 63% survival rate. Flow-cytometry based DNA content analysis revealed that the ploidy levels were maintained in in vitro regenerated plants. This is the first report for in vitro plant regeneration in E. ochreata.

Effect of cytokinin types, concentrations and their interactions on in vitro shoot regeneration of Chlorophytum borivilianum Sant. & Fernandez

Background Chlorophytum borivilianum is a rare medicinal plant originally distributed throughout the forest of India. The tubers of C. borivilianum are used as an aphrodisiac and impotence supplement. The propagation of C. borivilianum is possible through seeds and tubers, but conventional methods may take several months. Hence in vitro technique of shoot regeneration could be an efficient alternative means of propagating the species. Latest study reported microtuberization of C. borivilianum but there is no sufficient study on a rapid method for shoot multiplication and elongation. Results Young shoot buds of C. borivilianum were cultured on MS medium containing 6-benzylaminopurine (BAP) and Kinetin (Kn), both at 0, 8.88, 17.8 and 26.6 µM, either individually or in combinations. Proliferated shoots were subcultured on fresh medium of the same constituents on week 3 of culture for further shoot multiplication and elongation. BAP alone (8.88-26.6 µM) was significantly effective on shoot multiplication, while Kn alone (8.88-26.6 µM) was significantly effective on shoot elongation compared to the control containing MS basal medium without any plant growth regulator. However, combination of both cytokinins stimulated an interaction producing higher shoot number and shoot length compared to their individual application. Conclusions The most suitable combination was 8.88 µM BAP + 8.88 µM Kn, reaching a mean shoot number of 10.83 and shoot length of 6.85 cm.

Reguladores de crescimento vegetal no cultivo in vitro de Mentha x Piperita L / Plant growth regulators on in vitro culture of Mentha x piperita L

A micropropagação é uma técnica muitas vezes indicada para a multiplicação em larga escala de plantas com propriedades medicinais. Dentre elas, destaca-se a hortelã-pimenta (Mentha x Piperita L.), cujo óleo essencial é utilizado no tratamento de transtornos digestivos e respiratórios. Para otimizar o protocolo de micropropagação dessa espécie são necessários estudos, principalmente quanto à suplementação do meio de cultura para garantir a produção massal in vitro e posterior extração do óleo essencial. Nesse contexto, objetivou-se avaliar o efeito de concentrações e combinações de reguladores de crescimento vegetal na morfogênese in vitro de hortelã-pimenta. Segmentos nodais provenientes de plântulas estabelecidas in vitro foram utilizados como fonte de explante e inoculados em meio de cultura MS suplementado com 0; 2,0 e 4,0 mg L-1 de BAP (6-benzilaminopurina), 0; 0,5 e 1,5 mg L-1 de ANA (ácido naftaleno-acético) e 0; 0,5 e 1,0 mg L-1 de GA3 (ácido giberélico). O delineamento experimental adotado foi inteiramente casualizado, com os tratamentos distribuídos em esquema fatorial 3x3x3 com oito repetições. Concluiu-se que o BAP favoreceu a sobrevivência de segmentos nodais de M. x Piperita inoculados in vitro e, quando combinado ao GA3, promoveu a brotação dos explantes. Essas características, no entanto, não foram estimuladas pela adição de ANA ao meio de cultura. Conclui-se que após a multiplicação dos brotos in vitro estes devem ser transferidos para meio sem reguladores para seu desenvolvimento. Apesar dos efeitos benéficos do BAP na organogênese de M. x Piperita, elevadas concentrações deste regulador de crescimento vegetal promoveram a formação de calos.

Micropropagation is a technique used for the large-scale production of medicinal plants. Among them, peppermint (Mentha x piperita L.) may be mentioned because of the pharmacological importance of its essential oil, which is used on the treatment of digestive and respiratory disorders. Studies are needed in order to optimize the micropropagation protocol of this species, especially concerning the culture medium, to ensure the in vitro mass clonal production and to enable the future extraction of the plant essential oil. Therefore, this work aimed to evaluate the effects of concentrations and combinations of different plant growth regulators on the in vitro morphogenesis of peppermint. Nodal segments from plantlets already established in vitro were used as explants and inoculated on MS medium supplemented with 0, 2.0 and 4.0 mg L-1 of BAP (6-benzylaminopurine), 0, 0.5 and 1.5 mg L-1 of NAA (naphthalene acetic acid) and 0; 0.5 and 1.0 mg L-1 of GA3 (gibberellic acid). The experiment was in a completely randomized design, set up as a 3x3x3 factorial design with eight replicates. We concluded that BAP increases the survival rate of in vitro inoculated nodal segments of M. x piperita. In addition, its combination with GA3 stimulates explants shooting. Those aspects, however, are not promoted by the addition of NAA into the culture medium. Also, the results indicate that, after in vitro multiplication, peppermint shoots must be transferred to another medium without plant growth regulators for shoot elongation. High concentrations of BAP promote calli induction, despite having beneficial effects on the organogenesis of M. piperita.

Multiplicação de abacaxizeiros ornamentais em diferentes concentrações de bap e períodos de avaliação do desenvolvimento in vitro / Multiplication of ornamental pineapple in different concentrations of bap and periods evaluation of development in vitro

O mercado mundial de flores tem apresentando grande destaque econômico nos últimos anos. Apesar do estabelecimento das técnicas de micropropagação, melhorias no protocolo ainda são necessárias. Neste sentido, objetivou-se com o presente trabalho avaliar a multiplicação in vitro das espécies de abacaxi ornamental Ananas comosus var. bracteatus e A. comosus var. erectifolius em diferentes períodos de avaliação e concentrações de BAP. Foram utilizados como explantes gemas axilares de abacaxizeiro ornamental. Utilizou-se delineamento experimental inteiramente casualizado, composto de 16 tratamentos com 5 repetições e fatorial 2x4x2, sendo um explante por tubo de ensaio. Para o número de folhas produzidas, a variedade erectifolius foi mais eficiente que a bracteatus; na avaliação realizada aos 60 dias, o número de folhas foi 2 vezes superior aos 30 dias na concentração de 1 mg.L-1 de BAP. A variedade erectifolius apresentou folhas mais compridas na concentração de 1 mg.L-1 de BAP. O comprimento das folhas diminuiu com o aumento da concentração de BAP. O número de raízes produzidas não foi influenciado pela variedade, nem pelo período de avaliação. A variedade erectifolius apresentou maior comprimento de raiz principal que a bracteatus na ausência de BAP aos 60 dias. A concentração de 3 mg.L-1 de BAP foi mais eficiente na formação de brotos. A variedade bracteatus aos 60 dias apresentou a maior formação de brotos em abacaxizeiro ornamental.

The world market for flowers is showing great economic prominence in recent years. Despite the establishment of micropropagation techniques, improvements are still needed in the protocol. In this sense, the aim of the present study was to evaluate the in vitro multiplication of species of ornamental pineapple Ananas comosus var. bracteatus and A. comosus var. erectifolius at different evaluation periods and concentrations of BAP. Were used as explants axillary buds of ornamental pineapple. We used a completely randomized design, consisting of 16 treatments with 5 replicates and 2x4x2 factorial design, with one explant per tube. For leaf number, variety erectifolius was more efficient than bracteatus; evaluation performed 60 days, the number of leaves was two times higher than at 30 days at the concentration of 1 mg l-1 BAP. The variety erectifolius presented leaves longer in the concentration of 1 mg L-1 BAP. The length of the leaves decreased with increasing concentration of BAP. The number of roots produced was not influenced by the variety, or the evaluation period. The variety erectifolius showed greater root length main bracteatus that in the absence of BAP at 60 days. The concentration of 3 mg L-1 BAP was more effective in the formation of shoots. The variety bracteatus at 60 days showed the highest shoot formation in ornamental pineapple.

Giberelina, citocinina e auxina na qualidade química de bagas de uva 'Superior Seedless' / Gibberellin, citocinin and auxin on chemical quality of 'Superior Seedless' grape berries

Os experimentos foram conduzidos no município de Juazeiro/BA com a finalidade de avaliar o efeito de reguladores vegetais compostos por giberelina, citocinina e auxina na qualidade química de bagas de uva do cultivar Superior Seedless. No primeiro ensaio estudaram-se os efeitos de Stimulate (biorregulador) e X-Cyte (citocinina) associado a uma nova formulação comercial de giberelina (N-Large) e ao Pro-Gibb, produto usualmente utilizado como fonte de giberelina, aplicados via pulverização na fase de desenvolvimento das bagas (18, 21, 51 e 56 dias após a poda de produção). Para tanto se empregou os seguintes tratamentos: T1: Pro-Gibb; T2: Stimulate (Dose 1); T3: Stimulate (Dose 2); T4: Stimulate (Dose 3); T5: Pro-Gibb + X-Cyte (Dose Baixa - DB); T6: Pro-Gibb + X-Cyte (Dose Média - DM); T7: Pro-Gibb + X-Cyte (Dose Alta - DA); T8: N-Large; T9: N-Large + X-Cyte (DB); T10: N-Large + X-Cyte (DM); T11: N-Large + X-Cyte (DA). O segundo ensaio teve por objetivo verificar o efeito da nova formulação de giberelina (N-Large) associada ou não a citocinina (X-Cyte) também aplicados via pulverização direcionada aos cachos na fase de desenvolvimento das bagas (17, 55 e 66 dias após a poda de produção). Assim, os seguintes tratamentos foram empregados: T1: Pro-Gibb; T2: N-Large (DB); T3: N-Large (DM); T4: N-Large (DA); T5: N-Large (DB) + X-Cyte (DB); T6: N-Large (DB) + X-Cyte (DM); T7: N-Large (DB) + X-Cyte (DA); T8: N-Large (DM) + X-Cyte (DB); T9: N-Large (DM) + X-Cyte (DM); T10: N-Large (DM) + X-Cyte (DA); T11: N-Large (DA) + X-Cyte (DB); T12: N-Large (DA) + X-Cyte (DM); T13: N-Large (DA) + X-Cyte (DA). O delineamento experimental foi em blocos ao acaso com quatro repetições, onde se considerou cada parcela uma repetição com três plantas úteis na mesma linha de plantas. Por ocasião da colheita, momento em que a média dos cachos apresentava teor de sólidos solúveis superior a 15 ºBrix, foram retiradas bagas para as análises de sólidos solúveis, pH, acidez titulável e, cálculo do ratio (SS/AT). Em ambos os ensaios os reguladores vegetais avaliados não proporcionaram alterações significativas nas características químicas das bagas de uva 'Superior Seedless'. Assim, a ausência de diferença de resposta entre o produto comercialmente empregado (Pro-Gibb) e os demais testados (Stimulate, X-Cyte e N-Large) os torna promissores no cultivo da uva 'Superior Seedless', fornecendo maior número de alternativas aos produtores da região do Vale do São Francisco.

Trials were carried out in Juazeiro, Bahia State, Brazil, aiming to test plant regulators composed by gibberellin, cytokine and auxin effects on chemical quality of Superior Seedless grape berries. The first trial studied the effects of Stimulate (bio regulator) and X-Cyte (cytokine) associated to a new gibberellin formulation (N-Large ) and associated to Pro-Gibb , which is a product used as source of gibberellin. Products were sprayed at berries development phase (18, 21, 51 and 56 days after spur-pruning). Treatments were: T1: Pro-Gibb ; T2: Stimulate (Dose 1); T3: Stimulate (Dose 2); T4: Stimulate (Dose 3); T5: Pro-Gibb + X-Cyte (Low Dose - DB); T6: Pro-Gibb + X-Cyte (Intermediate Dose - DM); T7: Pro-Gibb + X-Cyte (High Dose - DA); T8: N-Large ; T9: N-Large + X-Cyte (DB); T10: N-Large + X-Cyte (DM); T11: N-Large + X-Cyte (DA). The second trial aimed to assess the effect of the new gibberellin formulation (N-Large ) associated or not with cytokine (X-Cyte ) also sprayed straight over the bunches at berries development phase (17, 55 e 66 days after spur-pruning). Treatments were: T1: Pro-Gibb - blank; T2: N-Large (DB); T3: N-Large (DM); T4: N-Large (DA); T5: N-Large (DB) + X-Cyte (DB); T6: N-Large (DB) + X-Cyte (DM); T7: N-Large (DB) + X-Cyte (DA); T8: N-Large (DM) + X-Cyte (DB); T9: N-Large (DM) + X-Cyte (DM); T10: N-Large (DM) + X-Cyte (DA); T11: N-Large (DA) + X-Cyte (DB); T12: N-Large (DA) + X-Cyte (DM); T13: N-Large (DA) + X-Cyte (DA). Experimental design was random blocks with four repetitions with each repetition/parcel having three useful plants in the same row. At harvest, when bunches average had soluble solids over 15 ºBrix, berries were collected for soluble solids, pH, titratable acidity analysis as well as (SS/AT) ratio calculation. In both trials, plant regulators evaluated did not provide significant changes on chemical quality of 'Superior Seedless' grape berries. Therefore, the lack of differences on response between the commercially used product (Pro-Gibb) and the other

Concentrações de BAP sobre a proliferação in vitro de brotos de Lippia alba [(Mill. )N. E. Brown] / BAP concentrations on in vitro proliferation of Lippia alba [(Mill. ) N. E. Brown] shoots

Lippia alba é uma planta de origem brasileira, pertencente à família Verbenaceae. É conhecida por combater a insônia e a asma, além de possuir ação anticonvulsivante e antifúngica. A micropropagação tem sido utilizada para multiplicação de várias espécies com propriedades medicinais, e o BAP é a citocinina sintética mais utilizada para a multiplicação in vitro. O objetivo deste trabalho foi estabelecer a concentração adequada de BAP na multiplicação in vitro de L. alba. Explantes constituídos de segmentos nodais provenientes de plântulas já estabelecidas in vitro com aproximadamente 0,5 cm foram inoculados em meio MS, suplementado com 0,0; 0,5; 1,0 e 1,5 mg L-1 de BAP em adição de 30 g L-1 de sacarose. A utilização de 1,5 mg L-1 de BAP promove a multiplicação in vitro de L. alba. A ausência deste regulador propicia maior número de folhas e o uso na concentração de 0,5 mg L-1 aumentou as massas fresca e seca de parte aérea nesta espécie.

Lippia alba is a Brazilian plant, belonging to the Verbenaceae family. It is known to combat insomnia and asthma, also has antifungal and anticonvulsant actions. Micropropagation has been used for propagation of various species with medicinal properties, and BAP is the most widely used synthetic cytokinin for in vitro multiplication. The aim of this study was to determine the appropriate concentration of BAP on in vitro multiplication of L. alba. Explants consisting of nodal segments from seedlings already established in vitro with approximately 0.5 cm were inoculated on MS medium supplemented with 0; 0.5; 1.0 and 1.5 mg L-1 of BAP and 30 g L-1 of sucrose. The use of 1.5 mg L-1 of BAP promotes the in vitro multiplication of L. alba. The absence of this regulator provides a greater number of leaves and its use at the concentration of 0.5 mg L-1 increased shoots' fresh and dry weights in this specie.

Cálcio e fitorreguladores no desenvolvimento e estado nutricional de Mentha spicata x suaveolens cultivada em solução nutritiva / Calcium and plant growth regulators on the development and nutritional status of Mentha spicata x suaveolens cultivated in nutrient solution

O trabalho teve como objetivo estudar o desenvolvimento e o estado nutricional de Mentha spicata x suaveolens cultivada em diferentes níveis de Calcio em solução nutritiva com aplicação de fitorreguladores. O experimento constou de esquema fatorial (4x4) sendo quatro níveis de Ca2+ na solução nutritiva (160, 120, 80 e 40 mg L-1) e três aplicações foliares, com intervalos de 15 dias, de ácido naftaleno acético (NAA), ácido giberélico (GA3), benzinaladenina (BA) e água destilada como testemunha. As plantas foram coletadas 60 dias após a transferência para a solução e avaliadas as variáveis, matéria fresca e seca, área foliar e número de folhas; volume, comprimento e superfície radiculares e teores de Ca, K, Mg, N e P das folhas. As variáveis foram submetidas à análise de variância e as médias comparadas pelo teste Tukey. Não foram encontradas diferenças entre os níveis de Ca para a produção de biomassa. A auxina não influenciou a matéria seca das plantas, embora tenha aumentado os teores foliares de Ca; a citocinina acarretou diminuição da matéria seca total e a giberelina aumento da matéria seca do caule. A variação de Ca2+ não influenciou os teores foliares de N, P, K e Mg. Houve resposta para aplicação de Ca2+, com teores foliares atingindo o máximo de 15,73 mg kg-1 na dose de 137,81 mg L-1 pela análise de regressão

This study aimed to verify the development and the nutritional status of Mentha spicata x suaveolens cultivated under different Ca2+ levels in nutrient solution with application of plant growth regulators. The experiment was in factorial design (4X4) with four Ca2+ levels in the nutrient solution (160, 120, 80 and 40 mg L-1) and three leaf applications, at 15-day intervals, of naphthalene acetic acid (NAA), gibberellic acid (GA3), benzyladenine (BA) and distilled water as control. Plants were harvested at 60 days after transferring to the nutrient solution and the following variables were evaluated fresh and dried matter, leaf area, number of leaves, volume, length and surface of roots, and leaf content of Ca, K, Mg, N and P. The variables were subjected to analysis of variance and means compared by Tukey's test. There were no differences between Ca levels for biomass production. The auxin did not influence the dry matter of plants although it increased the leaf content of Ca2+, the cytokinin reduced total dry matter and the gibberellin increased the stem dry matter. The change in Ca2+ did not influence the leaf content of N, P, K and Mg. There was response for the application of Ca2+, reaching the maximum of 15.73 mg L-1 leaf content at the level of 137.81 mg L-1, according to regression analysis

Citocininas na multiplicação in vitro de hortelã-pimenta (Mentha x Piperita L. ) / Cytokinins on in vitro multiplication of peppermint (Mentha x Piperita L. )

O hortelã-pimenta é um hibrido triplo, Mentha x piperita, utilizado de forma medicinal no tratamento de náuseas, cólicas gastrointestinais, flatulências, cálculos biliares, icterícia, ansiedade, expectoração e expulsão de vermes intestinais. Na micropropagação, o BAP (6-benzilaminopurina) e KIN (cinetina) têm sido as fontes de citocininas mais empregadas. O objetivo deste trabalho foi avaliar o efeito de reguladores vegetais na multiplicação in vitro de hortelã-pimenta. Explantes constituídos de segmentos nodais provenientes de plântulas já estabelecidas in vitro com aproximadamente 0,5 cm foram inoculados em meio MS, suplementado com diferentes associações de BAP e KIN, em adição de 30 g L-1 de sacarose. As concentrações de citocininas utilizadas foram 0,0; 1,0 e 2,0 mg L-1 de BAP e 0,0; 0,5 e 1,0 mg L-1 de KIN. O delineamento experimental foi o inteiramente casualizado, com nove tratamentos consistindo de cinco frascos cada um, sendo que cada frasco continha quatro explantes. A utilização de 2,0 mg L-1 de BAP promove a multiplicação in vitro de hortelã-pimenta, porém, diminui o índice de sobrevivência. O uso de citocininas aumenta a massa fresca e seca dos explantes e a ausência destes reguladores propicia o alongamento nesta espécie.

Peppermint is used for medical treatments of nausea, gastrointestinal cramps, flatulence, gallstones, jaundice, anxiety, sputum and expulsion of intestinal worms. In micropropagation, BAP (6-benzylaminopurine) and KIN (kinetin) have been the most widely used sources of cytokinins. The aim of this study was to evaluate the effect of plant growth regulators in multiplication of peppermint. Explants consisting of nodal segments from seedlings already established in vitro with approximately 0,5 cm were inoculated on MS medium supplemented with different combinations of BAP and KIN and 30 g L-1 of sucrose. The concentrations of cytokinins used were 0,0; 1,0 and 2,0 mg L-1 of BAP and 0,0; 0,5 and 1,0 mg L-1 of KIN. The experiment was set up in a completely randomized design with nine treatments consisting of five bottles each, and each vial contained four explants. The use of 2,0 mg L-1 of BAP promotes the in vitro multiplication of peppermint, but decreases the survival rate. Cytokinins increase fresh and dry weight of explants and the absence of these regulators provides the elongation in this specie.

Solasodine accumulation in regenerated plants of Solanum torvum Sw / Acúmulo de solasodina em plantas micropropagadas de Solanum torvum Sw

A nodal segment culture was developed in order to assess Solanum torvum Sw. regeneration and solasodine levels. The influence of auxins (indoleacetic acid, 1-Naphthaleneacetic acid) and benzyl adenine on S. torvum growth in micropropagation was investigated. A nodal segment culture was initiated with seeds germinated in MS basal medium added of GA3 and grown in different concentrations of IAA, IAA + BAP and NAA + BAP. Sixty-day-old plants from the in vitro culture were collected, frozen and lyophilized; then, the methyl orange method was used to quantify solasodine for the spectrophotometric assay. The best results regarding plant regeneration and solasodine accumulation were obtained by using the MS basal medium without addition of plant growth regulators; however, there was great production of calluses presenting friable bases. Based on these results, cell cultures can be initiated from such calluses with application of other auxins and cytokinins to enhance solasodine production, besides different elicitors, light intensities and sucrose concentrations.

A regeneração de Solanum torvum e a avaliação do conteúdo de solasodina foram os objetivos de cultura de segmentos nodais. A influência de auxinas (ácido 3-indolacético, ácido naftalenoacético) e de 6-benzilaminopurina no crescimento de S. torvum na micropropagação foi investigado. Cultura de segmentos nodais foi iniciada por sementes germinadas em meio básico MS acrescido de GA3 e cultivadas em diferentes concentrações de AIA, AIA + BAP e ANA + BAP. Plantas da cultura in vitro com 60 dias foram coletadas, congeladas e liofilizadas e o método de alaranjado de metila foi utilizado para quantificação de solasodina para o ensaio espectrofotométrico. Os melhores resultados para regeneração vegetal e acúmulo de solasodina foram alcançados no meio MS sem adição de reguladores de crescimento havendo, porém grande produção de calos de base friáveis. Esses resultados mostram que a partir desses calos, cultura de células pode ser iniciada com aplicação de outras auxinas e citocininas para o aumento da produção de solasodina além de diferentes eliciadores, intensidades luminosas e concentrações de sacarose.

Antisense expression of a gene encoding a calcium-binding protein in transgenic tobacco leads to altered morphology and enhanced chlorophyll.

Entamoeba histolytica contains a novel calcium-binding protein like calmodulin,which was discovered earlier,and we have reported the presence of its homologue(s)and a dependent protein kinase in plants.To understand the functions of these in plants,a cDNA encoding a calcium-binding protein isolated from Entamoeba histolytica (EhCaBP)was cloned into vector pBI121 in antisense orientation and transgenic tobacco plants were raised.These plants showed variation in several phenotypic characters,of which two distinct features,more greenness and leaf thickness,were inherited in subsequent generations.The increase in the level of total chlorophyll in different plants ranged from 60% to 70%.There was no major change in chloroplast structure and in the protein level of D1,D2,LHCP and RuBP carboxylase.These morphological changes were not seen in antisense calmodulin transgenic tobacco plants,nor was the calmodulin level altered in EhCaBP antisense plants.

Potential application of urea-derived herbicides as cytokinins in plant tissue culture.

Various urea-derived herbicides and different cytokinin analogues were used to determine their effects on callusing response and shoot regenerating capacity of alfalfa (Medicago sativa L.) and Coleus (Coleus forskohlii Briq.). The herbicides monuron and diuron evoked profuse callusing response from Coleus leaf segments and alfalfa petiole explants on Murashige and Skoog medium. Shoot regeneration by monuron (2.0 mg/l) showed a maximum of 3 multiple shoots both in alfalfa and Coleus with a frequency of 92% and 75%, respectively. Whereas diuron (0.5 mg/l) showed a high frequency of shoot regeneration (89%)with a mean number of 5 shoots in alfalfa, in C.forskohlii, the frequency of regeneration was 90%with a mean number of 6 shoots. Diuron with two chloride groups in the phenyl ring showed significantly higher cytokinin-like activity than single chloride substitution monuron. This study demonstrates the potential use of monuron and diuron as cytokinins in plant tissue culture.

Reduction of vitrification in in vitro raised shoots of Chlorophytum borivilianum Sant. & Fernand., a rare potent medicinal herb.

Reduction of vitrification in in vitro raised shoots derived from shoot bases and immature floral buds along with inflorescence axis used as explants of C. borivilianum, a rare medicinal herb is described. Shoot multiplication was obtained on MS medium with 2 mg l(-1) benzylaminopurine (BAP) + 0.1 mg l(-1) indole-3-butyric acid (IBA) and MS medium with 2 mg l(-1) kinetin (Kin) + 0.1 mg l(-1) 2,4-dichlorophenoxy acetic acid (2,4-D) from shoot bases and inflorescence axis respectively. Best multiplication rates were obtained from both the explants on MS medium with 2 mg l(-1) BAP. Vitrification of shoots in cultures appeared during the multiplication stage. Culture bottles with aerated caps reduced the vitrification to 80%. Reduction of BAP concentration from 2 mg l(-1) to zero during subsequent subcultures also minimized vitrification. Use of 0.5-2 mg l(-1) Kin produced healthy shoots when compared to BAP. In vitro raised shoots rooted on Knop salts containing iron and vitamins of MS medium, 2 mg l(-1) IBA and 0.1% activated charcoal. About 80% plantlets survived upon soil transfer. Scanning electron microscopic and image analyzer studies reveal the morphological structural differences between the leaves of normal and vitrified plantlets.

Influence of cytokinins, auxins and polyamines on in vitro mass multiplication of cotton (Gossypium hirsutum L. cv. SVPR2).

In the present investigation, the influence of different forms of cytokinins, auxins and polyamines were tested for mass multiplication and regeneration of cotton. Initially, for the identification of effective concentration for multiple shoot induction, various concentrations of BAP, Kin and 2iP along with IAA and NAA were tested. Among tested concentrations, media fortified with MS salts; B5 vitamins; 30 g/l, glucose; 2.0 mg/l, 2iP; 2.0 mg/l, IAA and 0.7 % agar showed best response for multiplication of shoot tip explants (20 shoots per shoot tip explants). In nodal explants, maximum of 18.6 shoots were obtained in the media fortified with MS salts, B5 vitamins, 30 g/l, glucose, 2.0 mg/l, 2iP, 1.0 mg/l, NAA and 0.7 % agar. Effect of different concentrations of polyamines like spermidine and putrescine were also tested along with the above said multiplication media. Among the various treatments, 20 mg/l of putrescine showed best response and the multiple of shoots were increased to 26.5 shoots per shoot tip explants and 24.5 shoots per nodal explants. Elongation of shoots was achieved on multiple shoot induction medium. Significant number of roots were initiated in the medium supplemented with MS salts, vitamin B5 and IBA (2.0 mg/l). The frequency of root induction was increased by addition of, PVP (10 mg/l) along with root induction medium and after 2 weeks, the roots reached the maximum length of 22 cm. Further, these plantlets were hardened by using sand, soil and vermiculate in 1:1:1 ratio. The hardened plants were transferred to the environmental growth chamber for proper acclimatization. The hardened plants were then transferred to field for boll yielding and they exhibited 100% survival.