Polyploidy induction in Physalis alkekengi / Indução de poliploidia em Physalis alkekengi

Physalis alkekengi is an ornamental plant that can also be used as a medicinal plant due to its anti-inflammatory, bactericidal, antitumor and fungicidal properties. Polyploidization can be an important tool in the genetic improvement of this species. The objective this work was to obtain tetraploids in vitro and to evaluate the phytotechnical traits of P. alkekengi. For this, nodal segments of P. alkekengi var. Franchettii were inoculated into petri dishes containing 100 ml of MS medium supplemented with colchicine at concentrations 0; 0.04; 0.08; 0.12; and 0.16% and kept in the dark for 24 and 48h. After the respective treatment periods with colchicine the segments were inoculated into test tubes. The tetraploids were identified by flow cytometry and classical cytogenetics. In vitro seedlings were measured: root length, nodal segment length, leaflet number and total leaf area. In the acclimatization phase, the area of the second leaf and total leaf, petiole radius, stem length, fruit weight with calyx, without calyx, fruit diameter, number of seeds and brix of the pulp were evaluated. Chlorophyll a, chlorophyll b, total chlorophyll, total carotenoid, total chlorophyll / total carotenoid ratio and chlorophyll a / b ratio were also estimated. The treatment that most produced tetraploid seedlings was with 0.08% colchicine per 24h. No significant difference was observed in 7 (seven) variables, these being all variables of photopigments, stem diameter (steam) and brix. In general, diploid (2x) plants were better in 9 (nine) while tetraploid seedlings were better in 6 (six) of the phytotechnical variables. It was concluded that the MS medium supplemented with 0.08% colchicine for 24 h allowed P. alkekengi tetraploides to be obtained with better phytotechnical qualities.

Physalis alkekengi é uma planta ornamental que também pode ser usada como planta medicinal devido às suas propriedades anti-inflamatórias, bactericidas, antitumorais e fungicidas. A poliploidização pode ser uma ferramenta importante para o melhoramento genético dessa espécie. O objetivo deste trabalho foi obter tetraplóides in vitro e avaliar as características fitotécnicas de P. alkekengi. Para isso, segmentos nodais de P. alkekengi var. Franchettii foram inoculados em placas de Petri contendo 100 ml de meio MS suplementado com colchicina nas concentrações 0; 0,04; 0,08; 0,12; e 0,16% e mantido no escuro por 24 e 48h. Após os respectivos períodos de tratamento com colchicina, os segmentos foram inoculados em tubos de ensaio. Os tetraplóides foram identificados por citometria de fluxo e citogenética clássica. As plântulas in vitro foram medidas: comprimento da raiz, comprimento do segmento nodal, número de folhetos e área foliar total. Na fase de aclimatação foram avaliadas a área da segunda folha e área foliar total, raio do pecíolo, comprimento do caule, peso do fruto com cálice, sem cálice, diâmetro do fruto, número de sementes e brix da polpa. Também foram estimadas clorofila a, clorofila b, clorofila total, carotenóides totais, razão clorofila total / carotenóide total e razão clorofila a / b. O tratamento que mais produziu mudas tetraplóides foi com colchicina a 0,08% por 24 horas. Não foi observada diferença significativa em 7 (sete) variáveis, sendo todas variáveis de fotopigmentos, diâmetro do caule (vapor) e brix. Em geral, as plantas diplóides (2x) foram melhores em 9 (nove) variáveis fitotécnicas, enquanto as mudas tetraplóides foram melhores em 6 (seis). Concluiu-se que o meio MS suplementado com colchicina a 0,08% por 24 h permitiu obter tetraploides de P. alkekengi com melhores qualidades fitotécnicas.

Natural products as new antimitotic compounds for anticancer drug development

Cell cycle control genes are frequently mutated in cancer cells, which usually display higher rates of proliferation than normal cells. Dysregulated mitosis leads to genomic instability, which contributes to tumor progression and aggressiveness. Many drugs that disrupt mitosis have been studied because they induce cell cycle arrest and tumor cell death. These antitumor compounds are referred to as antimitotics. Vinca alkaloids and taxanes are natural products that target microtubules and inhibit mitosis, and their derivatives are among the most commonly used drugs in cancer therapy worldwide. However, severe adverse effects such as neuropathies are frequently observed during treatment with microtubule-targeting agents. Many efforts have been directed at developing improved antimitotics with increased specificity and decreased likelihood of inducing side effects. These new drugs generally target specific components of mitotic regulation that are mainly or exclusively expressed during cell division, such as kinases, motor proteins and multiprotein complexes. Such small molecules are now in preclinical studies and clinical trials, and many are products or derivatives from natural sources. In this review, we focused on the most promising targets for the development of antimitotics and discussed the advantages and disadvantages of these targets. We also highlighted the novel natural antimitotic agents under investigation by our research group, including combretastatins, withanolides and pterocarpans, which show the potential to circumvent the main issues in antimitotic therapy.

Modulação da cicatrização na cirurgia do glaucoma após aplicação de acetato de triancinolona subconjuntival isolado ou em associação à mitomicina C: estudo experimental / Healing modulation in glaucoma surgery after application of subconjunctival triamcinolone acetate alone or combined with mitomycin C: an experimetal study

RESUMO Objetivo: estudar a eficácia e segurança do uso de acetato de triancinolona subconjuntival isolado ou em associação à mitomicina C como modulador da cicatrização de trabeculectomias em coelhos. Métodos: trinta coelhos machos, albinos, raça Nova Zelândia foram submetidos à trabeculectomia bilateralmente. Os animais foram divididos em quatro grupos experimentais com 15 olhos por grupo: controle, mitomicina C, acetato de triancinolona e acetato de triancinolona + mitomicina C. Tonometria de aplanação e análise clínica da bolha através do Sistema de Graduação de Moorfields foram obtidas no pós-operatório. Para a avaliação da cicatrização, procedeu-se à análise quantitativa do infiltrado inflamatório (polimorfonucleares) através da coloração Hematoxilina & Eosina e da proliferação vascular por imuno-histoquímica. Resultados: foi observada em todos os grupos diminuição significativa da pressão intraocular pós-operatória em relação à pré-operatória (p<0,001). Contudo, não houve diferença entre os grupos (p=0,186). O grupo acetato de triancinolona + mitomicina C apresentou melhores índices na altura máxima da bolha e na vascularização da área central da bolha (p=0,001); além disso, houve menor resposta inflamatória (p=0,001) e menor proliferação vascular (p=0,001) na fase intermediária do estudo em relação às monoterapias. Conclusão: a associação da mitomicina C ao acetato de triancinolona resultou numa ação sinérgica entre esses agentes, com bolhas mais amplas e difusas e menor infiltrado inflamatório e menor proliferação vascular em estágio intermediário do acompanhamento neste modelo animal.

ABSTRACT Objective: to study the efficacy and safety of the use of subconjunctival triamcinolone acetate alone or in combination with mitomycin C as a modulator of trabeculectomy healing in rabbits. Methods: we submitted thirty male, albino, New Zealand rabbits to bilateral trabeculectomy. We divided the animals into four experimental groups with 15 eyes per group: control, mitomycin C, triamcinolone acetate and triamcinolone acetate + mitomycin C. We performed aplanation tonometry and clinical analysis of the bleb through the Moorfields Graduation System in the postoperative period. For the evaluation of healing, we carried out the quantitative analysis of the inflammatory infiltrate (polymorphonuclear) through Hematoxylin & Eosin staining, and vascular proliferation, through immunohistochemistry. Results: we observed a significant decrease in postoperative intraocular pressure in all groups compared with the preoperative pressure (p<0.001). However, there was no difference between groups (p=0.186). The triamcinolone + mitomycin C acetate group presented better indices as for the maximum bleb height and vascularization of the bleb central area (p=0.001); in addition, there was a lower inflammatory response (p=0.001) and lower vascular proliferation (p=0.001) in the intermediate phase of the study compared with the monotherapies. Conclusion: the combination of mitomycin C and triamcinolone acetate resulted in a synergistic action between these agents, with broader and more diffuse blebs, less inflammatory infiltrate and less vascular proliferation in the intermediate stages of follow-up in this animal model.

Carcinoma espinocelular de conjuntiva com evolução para exenteração: relato de caso / Conjunctival squamous cell carcinoma requiring orbital exenteration: case report

Portador de carcinoma espinocelular da conjuntiva teve a lesão removida, com recorrência em outra localização. O paciente recebeu ciclos de 5-Fluoruracila como tratamento adjuvante à remoção cirúrgica, apresentando evolução desfavorável que chegou à exenteração orbitária. São feitos comentários quanto ao uso de antimitóticos no tratamento destas lesões.

A case of a squamous cell carcinoma of the conjunctiva is presented. The lesion was removed with recurrence in another conjunctival site. The patient received cycles of 5-Fluorouracil as adjuvant treatment to the surgical removal and presented unfavorable evolution, requiring orbital exenteration. Comments are made on the use of antimitotic drugs in the management of these lesions.

Arsenic trioxide induces depolymerization of microtubules in an acute promyelocytic leukemia cell line

BACKGROUND: Arsenic trioxide (As2O3) is a well-known and effective treatment that can result in clinical remission for patients diagnosed with acute promyelocytic leukemia (APL). The biologic efficacy of As2O3 in APL and solid tumor cells has been explained through its actions on anti-proliferation, anti-angiogenesis, and apoptotic signaling pathways. We theorize that As2O3 activates a pathway that disrupts microtubule dynamics forming abnormal, nonfunctioning mitotic spindles, thus preventing cellular division. In this study, we investigated how As2O3 induces apoptosis by causing microtubule dysfunction. METHODS: Cultured NB4 cells were treated with As2O3, paclitaxel, and vincristine. Flow cytometric analysis was then performed. An MTT assay was used to determine drug-mediated cytotoxicity. For tubulin polymerization assay, each polymerized or soluble tubulin was measured. Microtubule assembly-disassembly was measured using a tubulin polymerization kit. Cellular microtubules were also observed with fluorescence microscopy. RESULTS: As2O3 treatment disrupted tubulin assembly resulting in dysfunctional microtubules that cause death in APL cells. As2O3 markedly enhanced the amount of depolymerized microtubules. The number of microtubule posttranslational modifications on an individual tubulin decreased with As2O3 concentration. Immunocytochemistry revealed changes in the cellular microtubule network and formation of polymerized microtubules in As2O3-treated cells. CONCLUSION: The microtubules alterations found with As2O3 treatment suggest that As2O3 increases the depolymerized forms of tubulin in cells and that this is potentially due to arsenite's negative effects on spindle dynamics.

Search of new Antimitotics compounds from the Cuban flora / Búsqueda de nuevos compuestos con actividad antimitótica en la flora cubana

The main objective of anti-carcinogenic chemotherapy is to stop uncontrolled cellular proliferation. This has prompted us to begin a systematic survey of new effective inhibitors with ability to react with cytoskeletal components and arrest living, dividing cells. Even for traditional populations herbs-consuming, encouraging the use of species with chemopreventive actions could be helpful as part of life expectancy improvement strategies. Herbal products have significantly lower costs, exhibit little or no toxicity during long-term oral administration and are relatively available at large scale. Current work involved the screening of 85 extracts from Cuban medicinal plants, selected on the basis of traditional use, ethnobotanics and pharmacological information (antiparasitic, antitumour, abortive, etc.). Antitubulinic activity in the hydroalcoholics extracts was evaluated by using a modified version of the conventional turbidity assay of tubulin assembly/ disassembly. The activity limits of the news isolated antitubulin agents were thoroughly investigated. According to the presented results, the extracts displaying the highest antitubulinic activity were Tamarindus indica L., Lawsonia inermes L and Xanthium strumarium L.

Detener la proliferación celular es el principal propósito de la quimioterapia anticarcinogénica. Para ello se ha realizado una búsqueda a partir de fuentes naturales de nuevos inhibidores efectivos que reaccionen con los componentes del citoesqueleto y puedan detener la división celular. En poblaciones que tradicionalmente utilizan plantas medicinales se estimula el uso de aquellas especies con acción quimiopreventivas como parte de una estrategia que contribuya a la calidad de vida. Los productos herbarios tienen costos significativamente más bajos, exhiben poca o ninguna toxicidad durante la administración oral a largo plazo y están al alcance de todos. Nuestro trabajo consistió en realizar un tamizaje de 85 extractos de plantas medicinales cubanas seleccionadas en base al uso tradicional, en las encuestas etnobotánicas e información farmacológica (actividad antiparasitaria, antitumoral, abortiva, etc). La actividad antitubulínica fue evaluada mediante una versión modificada del ensayo turbimétrico del ensamblaje/desensamblaje de la tubulina. Se determinó la actividad límite de los nuevos agentes antitubulínicos siendo los extractos de Tamarindus indica L., Lawsonia inermes L and Xanthium strumarium L. los de mejor actividad antitubulínica según las condiciones ensayadas.

In vitro and in vivo antiproliferative activity of Calotropis procera stem extracts

The cytotoxic potential of stem organic extracts from Calotropis procera (Asclepiadaceae) was firstly evaluated against cancer cell lines by MTT assay. Subsequently, samples considered cytotoxic were tested for antimitotic activity on sea urchin egg development and for in vivo antiproliferative activity in mice bearing Sarcoma 180 tumor. Among the five extracts (hexane, dichloromethane, ethyl acetate, acetone and methanol), ethyl acetate and acetone extracts displayed higher cytotoxic potential against tumor cells, with IC50 ranging from 0.8 to 4.4 μg/mL, while methanolic extract was weakly cytotoxic. Cytotoxic extracts also exhibited cell division inhibition capacity by antimitotic assay, revealing IC50 values lower than 5 μg/mL. In the in vivo antitumor assessments, ethyl acetate- and acetone-treated animals showed tumor growth inhibition ratios of 64.3 and 53.1 percent, respectively, with reversible toxic effects on liver and kidneys. Further studies are in progress in order to identify C. procera cytotoxic compound(s) and to understand the mechanism of action responsible for this tumor-decreasing potential.

O potencial citotóxico de extratos orgânicos do caule de Calotropis procera (Asclepiadaceae) foi primeiramente avaliado frente a linhagens de células tumorais através do ensaio de MTT. Aquelas amostras consideradas citotóxicas foram sub-sequentemente testadas para atividade antimitótica sobre o desenvolvimento de ovos de ouriço-do-mar e para atividade antiproliferativa in vivo em camundongos transplantados com tumor Sarcoma 180. Dentre os cinco extratos estudados (hexano, diclorometano, acetato de etila, acetona e metanol), os extratos acetato de etila e acetona mostraram maior potencial citotóxico contra células tumorais, com CI50 variando de 0,8 to 4,4 μg/mL, enquanto o extrato metanólico revelou ser fracamente citotóxico. s extratos citotóxicos também exibiram capacidade de inibição da divisão celular com valores de CI50 menores que 5 μg/mL. Nas avaliações antitumorais in vivo, os animais tratados com os extratos acetato de etila e acetona mostraram taxas de inibição do crescimento tumoral de 64,3 e 53,1 por cento, respectivamente, com efeitos tóxicos reversíveis sobre o fígado e os rins.

Anti-mitotic activity towards sea urchin eggs of dichloromethane fraction obtained from Allamanda schottii Pohl (Apocynaceae) / Atividade anti-mitótica da fração diclorometano obtida de Allamanda schottii Pohl (Apocynaceae) sobre ovos do ouriço do mar

Allamanda (Apocynaceae) is a genus of climbing shrubs known for producing compounds with a range of biological activities. Previous works have shown the anti-proliferative effect of the ethanolic extract of Allamanda schottii on leukemic cells. The present work was conducted to evaluate the effects of dichloromethane fraction, obtained from Allamanda schottii, on sea urchin Echinometra lucunter eggs, as a multicellular model for evaluating anti-tumor activity. Our results show an inhibition of sea urchin development in a dose-dependent manner in the presence of dichloromethane fraction. The IC50 values for first and third cleavage and blastulae stage were 103.7 µg/mL, 33.1 µg/mL and 10.2 µg/mL, respectively. These results also demonstrate the cumulative effect of this fraction on sea urchin embryos. In the present work, the expressive anti-mitotic activity of dichloromethane fraction towards sea urchin eggs, a multicellular model, reinforces the anti-tumor potential of the Allamanda schotti.

Allamanda (Apocynacea) é um gênero de arbustos escandentes conhecido por produzir compostos com várias atividades biológicas. Trabalhos anteriores têm mostrado um efeito anti-proliferativo do extrato etanólico de Allamanda schottii sobre células leucêmicas. O presente trabalho foi realizado para avaliar o efeito da fração diclorometano, obtida de Allamanda schotti, sobre os ovos de ouriço-do-mar de Echinometra lucunter, como um modelo multicelular para estudar atividade anti-tumoral. Nossos resultados mostram uma inibição do desenvolvimento dos ovos de uma maneira dose-dependente na presença da fração diclorometano. Os valores de IC50 para a primeira e terceira clivagem e para o estágio de blástula foram de 103,7 µg/mL, 33.1 µg/mL e 10,2 µg/mL, respectivamente. Estes resultados também demonstram um efeito acumulativo da fração sobre os embriões do ouriço-do-mar. No presente trabalho, esta expressiva atividade anti-mitótica da fração diclorometano sobre o desenvolvimento embrionário do ouriço-do-mar, um modelo multicelular, reforça o potencial antitumoral de Allamanda schotti.

Segurança e efetividade no tratamento do pterígio usando infiltração de 5-fluoruracila no intraoperatório / Safety and efficacy of intraoperative 5-fluorouracil infiltration in pterygium treatment

Objetivo: Avaliar a segurança e a efetividade do uso do 5-fluoruracila(5-FU) como tratamento adjuvante do pterígio, aplicado sob a forma de infiltração subconjuntival, no período intraoperatório. Métodos: Foram avaliados prospectivamente 125 indivíduos (125 olhos) portadores depterígio. Os indivíduos foram operados segundo a técnica de retalho de deslizamento e receberam, ao final do procedimento, injeção subconjuntivalde 0,2 mL de 5-FU (25 mg/mL). Foram anotados os dados do paciente como idade, sexo, profissão, características da lesão (primário ou recidivado, tamanho, carnoso ou involutivo) e feito seguimento pósoperatório, aos 7, 21, 60 e 180 dias. Os dados foram submetidos àavaliação estatística. Resultados: Não foram observados casos de complicaçãodecorrente do uso do 5-FU em injeção no intraoperatório do pterígio. A taxa de recidiva geral observada aos 180 dias de pós-operatório foi de 35,8%, sendo de 35,7% para os pterígios primários e de 36,4% para os recidivados. Conclusão: A aplicação do 5-FU no período intraoperatório sob a forma de infiltração subconjuntival é segura. Entretanto, ainda resulta em altas taxas de recidiva e novos estudos devem serrealizados a fim de conhecer a concentração/dose ideal que permitirámenores chances de recidiva da lesão.

Purpose: To investigate the safety and efficacy of intraoperative infiltration of 5-fluorouracil (5-FU) as an adjuvant drug in pterygium treatment. Methods: Of 125 consecutive patients, 125 eyes with primary and recurrent pterygium underwent pterygium excision with intraoperative 5-FU (25 mg/mL) infiltration. The superior and inferior conjunctiva was approximated to cover the scleral bed and 0.2 mL 5-FU wasinjected at the end of the surgical procedure. The gender, occupation, pterygium characteristics and the follow-up at 7, 21, 60 e 180 days after surgery were evaluated and the data were statistically analyzed. Results: With follow-up of 180 days the patients had no serious complications observed duringor after surgery. The relapse rate was 35.8% and occurred in primary (35.7%) and recurrent (36.4%) lesions with no statistical difference. Conclusions: This study suggests that intraoperative infiltration of 5-FU is safe. However the high recurrence rate indicated that other studies would be necessary to show the concentration/dose to better prevent it.

Exposição de fibroblastos da cápsula de Tenon normal de portadores de pterígio ao 5-fluorouracil e à mitomicina C / Exposure of normal Tenon's capsule fibroblasts from pterygium to 5-fluorouracil and mitomycin C

OBJETIVO: Avaliar a atividade proliferativa dos fibroblastos da cápsula de Tenon normal, proveniente de portadores de pterígios primários e recidivados. MÉTODOS: Foi realizado estudo prospectivo, aleatório, avaliando-se fragmentos da cápsula de Tenon normal, removidos de 20 portadores de pterígios primários e 21, recidivados. A taxa de proliferação foi avaliada em fibroblastos de terceira passagem, quando as culturas foram expostas a agentes antimitóticos: mitomicina C e 5-fluorouracil. Os dados obtidos foram submetidos à análise estatística. RESULTADOS: Dentre os 41 espécimes cultivados, apenas 1 de pterígio primário e 2 de recidivado proliferaram. Quando expostos a mitomicina C e ao 5-fluorouracil não houve diferença estatisticamente significativa quanto a inibição de proliferação celular. CONCLUSÃO: Desta forma, in vitro, ambos os antimitóticos estudados têm a mesma eficácia na inibição da proliferação sobre os fibroblastos de cápsulas de Tenon normal.

PURPOSE: To evaluate the fibroblast proliferation activity of normal Tenon's capsule from primary and recurrent patients with pterygium. METHODS: A randomized prospective study was performed with 41 normal Tenon's capsule fragments from 21 primary and 20 recurrent patients with pterygium. The sample was collected from the inferior cul-de-sac. Proliferation rate from fibroblasts were evaluated after mitomycin C and 5-fluorouracil exposition. Data were submitted to statistical analysis. RESULTS: Of the 41 cultivated normal Tenon's capsules, only 1 from primary and 2 from recurrent pterygium patients proliferated. After antimitotic exposition, the proliferation rate was similar with both drugs. CONCLUSION: Mitomycin and 5-fluorouracil promote similar inhibition regarding proliferation of normal Tenon's fibroblast cultures.

Effects of topical application of 5-FU on the corneoscleral limbus of rabbits / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To observe the effects of topical application of 5-fluorouracil(5-FU)on the rabbits corneal limbus.METHODS:5-FU was applied topically to the corneoscleral region of rabbits eyes.The animals were sacrificed immediately or 7,15,30 and 60 days later,and tissues were submitted to histological examination.RESULTS:All animals presented corneosclerai deepithelization close to the site of application during the immediate postoperative period,whereas on the 4th postoperative day ulceration was no longer present.Histological examination showed absence of epithelium and slight edema.After one week(G2),2 animals presented epithelial defects,thickened epithelium with Iarger basal cells and loose chromatin,and slight subepithelial edema.The remaining groups showed no alterations.CONCLUSION:The 5-FU topically on the corneoscleral limbus Dostpone the epitelization.

A Case of Therapy-related Acute Myeloid Leukemia associated with inv(16) / 대한진단검사의학회지

The inv(16)(p13q22) is found in de novo AML and is closely associated with the FAB subtype M4eo. The inv(16) is rarely reported in therapy-related AML (t-AML) patients. Herein, we report a case of t-AML with inv(16) after combination chemotherapy using antimitotic agent and alkylating agent (cis-platin-paclitaxel) for ovarian serous cystadenocarcinoma.