Regeneración de plantas de té (Camellia sinensis) por cultivo in vitro de meristemas, yemas axilares y segmentos uninodales

Tres tipos de explantes de dos clones ( C H 1 4 I N TA y C H 3 1 8 I N TA ) d e t é (Camellia sinensis (L.) O. Kuntze) fueron evaluados para su regeneración in vitro, bajo la influencia de dos citocininas (BAP y CIN) y una giberelina (AG3). Previa desinfección, con etanol 70% (1 minuto) e hipoclorito de sodio 1,5% (20 minutos) y tres enjuagues con agua destilada estéril, los explantes fueron aislados y cultivados en los distintos medios de cultivo. Las mejores respuestas en formación de vástagos se registraron con los segmentos uninodales de ambos clones cultivados en el medio ½ MS + 1 mg/L de BAP o con el cultivo de yemas axilares del clon CH 14 INTA en el medio ½ MS + 1 mg/L de BAP o del clon CH 318 INTA en el medio ½ MS + 1 mg/L BAP + 1 mg/L AG3. Los mejores resultados con el empleo de meristemas caulinares se obtuvieron en el medio ½ MS + 1 mg/L de CIN y 1 mg/L de AG3. Los vástagos obtenidos fueron enraizados mediante su cultivo en ¼ MS + 6 mg/L de IBA.

Evaluation of molar teeth and buds in patients submitted to mandible distraction: Long-term results

Background: Despite all benefits offered by mandible distraction, complications and long-term consequences need to be evaluated to define its safety and morbidity. Forty mandible distractions were studied. Panoramic mandible radiographs obtained preoperatively, during distraction, and during the postoperative period were reviewed, with the intention of evaluating development and complications of molar buds and teeth in the distraction area. Methods: The mean patient age was 8.1 years. Twenty-five patients had craniofacial microsomia (one associated with a no. 10 facial cleft), five had temporomandibular joint ankylosis, two had familiar cases of auriculocondylar syndrome, one had a Tessier no. 30 facial cleft, and one had Treacher Collins syndrome. The severity of mandible hypoplasia was Pruzansky grade I in four cases, grade IIA in eight cases, grade 1113 in 16 cases, and grade III in one case. Mean radiographic follow-up was 44.8 months. Results: Molar buds located in the distraction area erupted without any deformity or displacement in 18 sides (45 percent). Fourteen cases presented distalization of a dental bud to a superior position in the mandibular ramus (four migrated back to the original position). Six molar buds presented perforations, four had shape deformities (two caused by dental fracture), and two had dental root injuries followed by root absorption lately. One case developed a dentigerous cyst. Conclusions: Almost half of the patients did not have any molar bud or tooth alterations after mandible distraction, and more than 20 percent presented only bud distalization. Therefore, preventive bud enucleation or tooth extraction should be avoided before mandible distraction.

Rapid increases in cytokinin concentration in lateral buds of chickpea (Cicer arietinum L) during release of apical dominance

We examined the role of cytokinins (CKs) in release of apical dominance in lateral buds of chickpea (Cicer arietinum L.). Shoot decapitation or application of CKs (benzyladenine, zeatin or dihydrozeatin) stimulated rapid bud growth. Time-lapse video recording revealed growth initiation within 2 h of application of 200 pmol benzyladenine or within 3 h of decapitation. Endogenous CK content in buds changed little in the first 2 h after shoot decapitation, but significantly increased by 6 h, somewhat later than the initiation of bud growth. The main elevated CK was zeatin riboside, whose content per bud increased 7-fold by 6 h and 25-fold by 24 h. Lesser changes were found in amounts of zeatin and isopentenyl adenine CKs. We have yet to distinguish whether these CKs are imported from the roots via the xylem stream or are synthesised in situ in the buds, but CKs may be part of an endogenous signal involved in lateral bud growth stimulation following shoot decapitation. To our knowledge, this is the first detailed report of CK levels in buds themselves during release of apical dominance.

Sox18 expression in blood vessels and feather buds during chicken embryogenesis

Sox18 encodes a transcription factor known to be important for the development of blood vessels and hair follicles in mice. In order to study the functional conservation of this gene through evolution, we have isolated and characterized Sox18 in chickens. cSox18 shows a high degree of sequence homology to both the mouse and human orthologues, particularly in the high mobility group DNA-binding domain and to a lesser extent in the transcriptional activation domain. A region of unusually high sequence conservation at the C-terminus may represent a further, previously unrecognized functional domain. Both the chicken and human proteins appear to be truncated at the N-terminus relative to mouse SOX18. In situ hybridization analyses showed expression in the developing vasculature and feather follicles, consistent with reported expression in the mouse embryo. In addition, cSox18 mRNA was observed in the retina and claw beds. (C) 2001 Elsevier Science B.V. All rights reserved.

XRCC3 THR241MET polymorphism influence on nuclear buds frequency in exposed to formaldehyde

Formaldehyde (FA), also known as formalin, formal and methyl aldehydes, is a colorless, flammable, strong-smelling gas. It has an important application in embalming tissues and that result in exposures for workers in the pathology anatomy laboratories and mortuaries. Occupational exposure to FA has been shown to induce nasopharyngeal cancer and has been classified as carcinogenic to humans (group 1) on the basis of sufficient evidence in humans and sufficient evidence in experimental animals. Manifold in vitro studies clearly indicated that FA is genotoxic. FA induced various genotoxic effects in proliferating cultured mammalian cells. The cytokinesis-block micronucleus (CBMN) assay was originally developped as an ideal system form easuring micronucleus (MN), however it can also be used to measure nucleoplasmic bridges (NBP) and nuclear buds (NBUD). Over the past decade another unique mechanism of micronucleus formation, known as nuclear budding has emerged. NBUDS is considered as a marker of gene amplification and/or altered gene dosage because the nuclear budding process is the mechanism by which cells removed amplified and/excess DNA.

Genotoxicity biomarkers micronucleus, nucleoplasmic bridges and nuclear buds

The increase of mortality from cancer brought urgency in identification and validation of predictive markers of risk and therefore early diagnosis. There is evidence that cytogenetic biomarkers are positively correlated with risk of cancer, and this is validated by studies of cohort and case-control. Cytokinesis-blocked micronucleus (CBMN) assay is used extensively in molecular epidemiology, and can be considered as a “cytome” assay covering cell proliferation, apoptosis, necrosis and chromosomal changes. The chromosomal alterations most reported and studied by the CBMN are: micronucleus (MN), nucleoplasmic bridges (NPB) and nuclear buds (NBUDS). The use of the MN assay in biomonitoring studies had a large increase in the last 15 years and international projects such as the HUMN have helped to increase the applicability and reliability of these tests.

Mechanical damage in cotton buds caused by the boll weevil

The boll weevil (Anthonomus grandis Boheman) causes high levels of bud abscission in cotton plants due to feeding or oviposition punctures. It has been reported that abscission is mainly due to enzymes present in the insect's saliva, but mechanical damage could also contribute to square abscission. The objective of this paper was to undertake an analysis of the morphological damages caused by the insect in cotton squares using microscopy. Anthers and ovules are the main target of boll weevil feeding. The process initiates by perforation of young sepal and petal tissues and proceeds with subsequent alimentation on stamen and ovary leading to abscission of floral structures.

Gray mold severity and vase life of rose buds after pulsing with citric acid, salicylic acid, calcium sulfate, sucrose and silver thiosulfate

Gray mold of roses (Rosa hibrida) caused by Botrytis cinerea requires many management strategies for its control. The effect of pulsing rose cv. Kiss with solutions of citric acid, salicylic acid, sucrose, calcium sulfate, and silver thiosulfate (STS) on disease severity and vase life of the flowers was evaluated. The solutions were applied to cut stems at different stages of harvest, the variation in the opening stage of harvest did not affect the results. Pulsing with STS reduced the values of area under the disease progress curve (AUDPC) and of severity of disease by 15% and 55%, respectively, and increased the vase life of the flowers by 20%. Calcium sulfate consistently reduced AUDPC by 66% and maximum severity by 88%, and increased vase life of the flowers by 37%. Therefore, pulsing rose buds with solutions of STS and calcium sulfate is potentially useful in reducing losses due to gray mold after harvest and in extending the vase life.

Cryopreservation of winter-dormant apple buds: 1 - variation in recovery with cultivar and winter conditions

The widely-adopted protocol for the cryopreservation of winter buds of fruit trees, such as Malus and Pyrus, was developed in a region with a continental climate, that provides relatively hard winters with a consequent effect on adaptive plant hardiness. In this study the protocol was evaluated in a typical maritime climate (eastern Denmark) where milder winters can be expected. The survival over two winters was evaluated, looking at variation between seasons and cultivars together with the progressive reduction in survival due to individual steps in the protocol. The study confirms that under such conditions significant variation in survival can be expected and that an extended period of imposed dehydration at -4oC is critical for bud survival. The occurrence of freezing events during this treatment suggests that cryodehydration may be involved, as well as evaporative water loss. To optimize the protocol for maritime environments, further investigation into the water status of the explants during cryopreservation is proposed. Keywords: Malus x domestica, cryopreservation, dormant bud, survival, grafting

Cryopreservation of winter-dormant apple buds: II - tissue water status after desiccation at -4°c and before further cooling

Abstract The established protocol for the cryopreservation of winter-dormant Malus buds requires that stem explants, containing a single, dormant bud are desiccated at -4°C, for up to 14 days, to reduce their water content to 25-30% of fresh weight. Using three apple cultivars, with known differences in response to cryopreservation, the pattern of evaporative water loss has been characterised, including early freezing events in the bud and cortical tissues that allow further desiccation by water migration to extracellular ice. There were no significant differences between cultivars in this respect or in the proportions of tissue water lost during the desiccation process. Differential Scanning Calorimetry (to -90°C) of intact buds indicated that bud tissues of the cultivar with the poorest response to cryopreservation had the highest residual water content at the end of the desiccation process and froze at the highest temperature Keywords: Malus, cryopreservation, dormant bud, dehydration

FIG CLONES SELECTION OF cv. ROXO DE VALINHOS OBTAINED BY IRRADIATED BUDS

The fig tree (Ficus carica L.) is a fruit tree of great world importance and, therefore, the genetic improvement becomes an important field of research for the crop improvement, being necessary to gather information on this species, mainly regarding its genetic variability so that appropriate propagation projects and management are made. However, the fig, in Brazil, is all produced from only one cultivar, Roxo de Valinhos, which produces seedless fruit, making impossible the conventional breeding. So, the fig breeding through induced mutagenic becomes a very important research line, greatly contributing to the fig culture development. The objective of this study was to select fig plants formed by cuttings treated with gamma ray. The plants used were obtained from buds of the cv. Roxo de Valinhos. The cuttings were irradiated with gamma rays in an irradiator Gamma Cell at 10 cm from the tip of the cutting, at doses of 30 Gy with dose rate of 238 Gy/h. The experiment consisted of 450 treatments, where each formed plant was a treatment. The treatments were numbered sequentially from 1 to 450 and spaced 2.5 x 1.5 m. It was evaluated the vegetative and the fruits characteristics, and the incidence of major crop pests and diseases. The analysis data showed that there is genetic variability among treatments and that the plants under numbers 1, 5, 20, 79, 164, 189, 194, 201, 221, 214, 258, 301, 322, 392, 433 and 440 are probably genetic mutants that should be tested as commercial orchards.

DAPK loss in colon cancer tumor buds: implications for migration capacity of disseminating tumor cells.

Defining new therapeutic strategies to overcome therapy resistance due to tumor heterogeneity in colon cancer is challenging. One option is to explore the molecular profile of aggressive disseminating tumor cells. The cytoskeleton-associated Death-associated protein kinase (DAPK) is involved in the cross talk between tumor and immune cells at the invasion front of colorectal cancer. Here dedifferentiated tumor cells histologically defined as tumor budding are associated with a high risk of metastasis and poor prognosis. Analyzing samples from 144 colorectal cancer patients we investigated immunhistochemical DAPK expression in different tumor regions such as center, invasion front, and buds. Functional consequences for tumor aggressiveness were studied in a panel of colon tumor cell lines using different migration, wound healing, and invasion assays. DAPK levels were experimentally modified by siRNA transfection and overexpression as well as inhibitor treatments. We found that DAPK expression was reduced towards the invasion front and was nearly absent in tumor buds. Applying the ECIS system with HCT116 and HCT116 stable lentiviral DAPK knock down cells (HCTshDAPK) we identified an important role for DAPK in decreasing the migratory capacity whereas proliferation was not affected. Furthermore, the migration pattern differed with HCTshDAPK cells showing a cluster-like migration of tumor cell groups. DAPK inhibitor treatment revealed that the migration rate was independent of DAPK's catalytic activity. Modulation of DAPK expression level in SW480 and DLD1 colorectal cancer cells significantly influenced wound closure rate. DAPK seems to be a major player that influences the migratory capability of disseminating tumor cells and possibly affects the dynamic interface between pro- and anti-survival factors at the invasion front of colorectal cancer. This interesting and new finding requires further evaluation.

Expression of Sonic hedgehog gene in regenerating newt limb blastemas recapitulates that in developing limb buds

This study aimed at characterizing the Sonic hedgehog (shh) gene in newt limbs, which encodes a signaling molecule of the zone of polarizing activity (ZPA) responsible for determining the anterior–posterior axis of the embryonic chicken and mouse limbs. The reverse transcription–PCR showed that adult newt regenerating limbs express shh genes. In situ hybridization experiments demonstrated that shh genes were expressed in mesenchymal cells of the posterior region of both embryonic buds and regenerating blastemas of newt limbs, strongly suggesting the presence of ZPA in these tissues. Experiments of the axial reversal graft of blastemas further supported this suggestion. The grafted blastemas regenerated supernumerary limbs, and this has been explained by three models: the polar coordinate model, the boundary model, and the polarizing zone model. In favor of the third model, the shh gene was expressed not only in the original region (new anterior region) of the graft, but also ectopically in the other region (new posterior region) of the same graft. This study implies that the regenerating limb blastema produces ZPA as the signaling center of the AP patterning as in the developing limb bud and, therefore, supports the notion that the limb regeneration recapitulates the limb development.

The Golgi-associated COPI-coated buds and vesicles contain β/γ-actin

It has been shown previously that the morphology and subcellular positioning of the Golgi complex is controlled by actin microfilaments. To further characterize the association between actin microfilaments and the Golgi complex, we have used the Clostridium botulinum toxins C2 and C3, which specifically inhibit actin polymerization and cause depolymerization of F-actin in intact cells by the ADP ribosylation of G-actin monomers and the Rho small GTP-binding protein, respectively. Normal rat kidney cells treated with C2 showed that disruption of the actin and the collapse of the Golgi complex occurred concomitantly. However, when cells were treated with C3, the actin disassembly was observed without any change in the organization of the Golgi complex. The absence of the involvement of Rho was further confirmed by the treatment with lysophosphatidic acid or microinjection with the constitutively activated form of RhoA, both of which induced the stress fiber formation without affecting the Golgi complex. Immunogold electron microscopy in normal rat kidney cells revealed that β- and γ-actin isoforms were found in Golgi-associated COPI-coated buds and vesicles. Taken together, the results suggest that the Rho signaling pathway does not directly regulate Golgi-associated actin microfilaments, and that β- and γ-actins might be involved in the formation and/or transport of Golgi-derived vesicular or tubular intermediates.

Interleaflet clear space is reduced in the membrane of COP I and COP II-coated buds/vesicles.

Intracellular transfers between membrane-bound compartments occur through vesicles that bud from a donor compartment to fuse subsequently with an acceptor membrane. We report that the membrane that delimits COP I or COP II-coated buds/vesicles from the endoplasmic reticulum and the Golgi complex has a thinner interleaflet clear space as compared with the surrounding, noncoated parental membrane. This change is compatible with a compositional change of the membrane bilayer during the budding process.