Hexaploid wheat genotypes from north-western Europe show low responses to current anther culture techniques. This phenomenon was investigated on 145 north-western European wheat lines. Twenty-seven lines from eastern Europe were included to observe the response pattern of wheat from an area, where the technique has been used successfully. On average, eastern European wheat lines produced 3.6 green plants per 111 anthers, while only 1.4 green plants per 111 anthers were obtained in north-western European lines. This difference was due to the high capacity for embryo formation among the eastern European lines, while the ability to regenerate green plants was widespread in both germplasm groups. Isolated wheat microspore culture performed on 85 of these wheat lines gave an average 3.7-fold increase in green plants per anther compared with the anther culture response. The increased recovery of green plants was due to improved plant regeneration and increased green plant percentage from embryos derived from isolated microspore culture

We studied reversible anther opening in a dichogamous aquatic plant Butomus umbellatus L. (Butomaceae) to assess its consequence on male fitness. Light microscope observations indicated that stomium breakage was simultaneous in all the anthers within an opening flower; however, detachment of the epidermis and outward bending of the anther wall were asynchronous. SEM observations showed that epidermis cells alternated between orbicular and crinkly shapes in response to absorption and loss of water. This generated centripetal and centrifugal forces which were significant enough to cause inward and outward bending of the anther wall, thus causing opened anthers to close and closed anthers to re-open respectively, depending on relative humidity of the environment. Behaviour of in vitro and in ...

Optimization of the isolation technique and initiation culture medium are two critical aspects that can determine the success of anthurium half-anther culture. Both aspects in half-anther culture of Anthurium andreanum Linden ex Andr? cv. ?Tropical? were studied and successfully improved. Untreated half-anthers, when cultured abaxial side down on medium, was the most suitable means of inducing callus. Callus formation was further improved by culturing half-anthers adaxial side down on Winarto-Teixeira (WT) medium (Winarto et al. in Plant Growth Regul 65:513?529, 2011b) supplemented with 0.01?mg/l ?-naphthaleneacetic acid (NAA), 1.0?mg/l 6-benzyladenine (BA) and 0.5?mg/l thidiazuron (TDZ). Gelrite enhanced callus formation (compared to agar) when the concentration was reduced from 2.0 to 1....

Abstract in portuguese A aveia (Avena spp.) tem sido pouco responsiva à haplodiploidização, um processo que aumenta a eficiência da seleção no melhoramento por gerar, em uma etapa, linhas puras homozigóticas. A fase mononucleada do micrósporo é critica para o sucesso da androgênese in vitro nos cereais de inverno e, em geral, pode ser inferida pelo tamanho da antera. Foram medidas anteras e analisados citológicamente micrósporos das cultivares de Avena sativa UPF 7, UPF 18, UFRGS 14 (more) , Stout e da linhagem CAV 3361 de Avena sterilis, cultivadas em câmaras de crescimento sob temperaturas dia-noite variando de 16ºC a 9ºC e 12 horas de intensidade luminosa de 300 mol m-2 s-1. O tamanho das anteras em cada fase de desenvolvimento dos micrósporos variou significativamente entre genótipos e de acordo com a região de inserção das espiguetas na panícula. A variação na arquitetura da panícula e a maturação não linear das espiguetas aumentam as dificuldades para a identificação das anteras potencialmente androgenéticas e podem explicar, em parte, os baixos resultados da androgênese na aveia. Os dados mostram a necessidade de uma análise citológica prévia para auxiliar a determinar a fase ideal de desenvolvimento dos micrósporos potencialmente responsivos à cultura de anteras, para o uso da androgenese na aveia. Abstract in english Oat (Avena spp.) is poorly responsive to the haplodiploidization process, which leads to the production of homozygous lines in one step, increasing breeding efficiency. Androgenetic haploids in small grain cereal crops are obtained from microspores cultured at the mononucleate stage, which can be identified by the size of anthers. In order to identify the appropriate anther size for in vitro culture, microspore cytological analyses were made in Avena sativa cultivars UPF (more) 7, UPF 18, UFRGS 14, Stout and Avena sterilis CAV 3361, cultivated in growth chamber under controlled light and temperature conditions. Variation was observed within and among genotypes for anther size at each microspore developmental stage and according to the position of spikelets in the panicle. Architecture variation in panicle shape and non-linear microsporogenesis maturation increased the challenge of identifying potentially androgenetic oat anthers. Cytological screening before culture is critical in identifying microspores at the right stage for oat androgenesis.

This report describes advances in the anther culture of Zantedeschia aethiopica. Important factors for improvement as compared to the earlier procedure were: (1) using flowers from inflorescences developed at relatively low temperature during winter, (2) high temperature stress treatment at 32^oC for 2 days in the beginning of the culture, (3) use of Gamborg B5 as anther culture medium, and (4) addition of sucrose at high concentration of 8% in the culture medium. Plants were obtained via a callus phase. Frequency of anthers producing calli was around 4-5%. About 87% of the calli gave regenerants, of which 52% were haploid, 36% were diploid and the rest had other ploidy levels. In addition to chromosome counting, cytological examination of the microspore development and amplified fragment ...

Anthurium anther culture was successfully established using half-anthers as explants. Explants were cultured on Winarto?Teixeira basal medium (WT-1) containing 0.01?mg/l ?-naphthalene acetic acid (NAA), 0.5?mg/l thidiazuron (TDZ), and 1.0?mg/l 6-benzylaminopurine (BAP), or on New Winarto?Teixeira basal medium (NWT-3) supplemented with 0.02?mg/l NAA, 1.5?mg/l TDZ, and 0.75?mg/l BAP for callus initiation. Regenerated calli produced multiple shoots on WT-1, which were then rooted in NWT-3 supplemented with 1% activated charcoal. Plantlets were acclimatized ex vitro using a mixture of burned rice husk, rice husk, and bamboo peat (1:1:1, v/v/v) as the potting medium. There was considerable morphological and cytological diversity of regenerants derived from anther culture, which are described in...

Transgenic rice was developed from both calli and immature embryos of popular cultivar ‘Swarna’ with a rice chitinase gene (chi11) by particle gun bombardment. Homozygous dihaploid transgenics were produced in less than a year through the anther culture of primary transgenics. Stable integration and expression of the chi11 transgene were confirmed by Southern and western analyses, respectively of primary as well as anther culture-derived dihaploid transgenics. The homozygous transgenics with functional transgene and varied levels of chitinase activity showed enhanced resistance to sheath blight fungus. The bioassay data were correlated with the molecular and biochemical results.   

http://www.springerlink.com/content/688481281g18035q/fulltext.pdf | In cereals, albinism is a major obstacle to produce doubled haploids (DH) for breeding programs. In order to identify QTLs for green plant percentage in barley anther culture, a specific populat...

The past 30 years have witnessed a series of systematic biotechnological advances made in pomegranate. These encompass optimization and establishment of in vitro culture techniques including micropropagation, somatic embryogenesis, synthetic seed production, plant regeneration via callus-mediated shoot organogenesis, adventitious shoot regeneration, anther culture, tetraploid induction and genetic transformation. This review attempts to provide a comprehensive account on the tissue culture-mediated biotechnological interventions made in pomegranate aimed at complementing conventional programmes for improvement of this nutraceutically important fruit crop. PMID:21161233

Protocols were developed for the generation of haploid or doubled haploid plants from developing microspores and ovules of Gentiana triflora. Plant regeneration was achieved using flower buds harvested at the mid to late uninucleate stages of microspore development and then treated at 4?C for 48?h prior to culture. Anthers and ovaries were cultured on modified Nitsch and Nitsch medium supplemented with a combination of naphthoxyacetic acid and benzylaminopurine. The explants either regenerated new plantlets directly or produced callus that regenerated into plantlets upon transfer to basal media supplemented with benzylaminopurine. Among seven genotypes of different ploidy levels used, 0?32.6% of cultured ovary pieces and 0?18.4% of cultured anthers regenerated plants, with all the genotype...

  The new brassinosteroid conjugate, teasterone-3-O-?-D-glucopyranoside, was found as a metabolite of teasterone in lily cell suspension cultures. Its structure was determined by means of FAB-MS and 1H-NMR upon comparison with the authentic compound. Furthermore, its presence in lily anthers was confirmed by FAB-MS and LC-APCI-SIM data. This is the first natural brassinosteroid conjugate glucosylated at a hydroxyl group in ring A.   

Salinity is a major factor that limits rice production worldwide. Rice is considered generally to be sensitive to salt stress during the reproductive stage. To determine the molecular mechanisms of salt tolerance at the reproductive stage, anther proteomic patterns for two contrasting rice genotypes IR64 (salt sensitive) and Cheriviruppu (salt tolerant) under salt stress were compared. Plants were grown in a greenhouse and salt stress (100 mM NaCl) was imposed at the booting stage. Anther samples were collected from control and salt-treated plants at the anthesis stage. The Na(+)/K(+) ratio in IR64 anthers under salt stress was >1.7 times greater than that under control conditions, whereas no significant change was observed in Cheriviruppu. We also observed an 83% reduction in IR64 pollen viability, whereas this reduction was only 23% in Cheriviruppu. Of 454 protein spots detected reproducibly on two-dimensional electrophoresis gels, 38 showed significant changes in at least one genotype in response to stress. Using Mass spectrometry (MALDI TOF/TOF) analysis, we identified 18 protein spots that were involved in several processes that might increase plant adaptation to salt stress, such as carbohydrate/energy metabolism, anther wall remodelling and metabolism, and protein synthesis and assembly. Three isoforms of fructokinase-2 were upregulated only in Cheriviruppu under salt stress. This upregulation might result in increased starch content in pollen, which would support pollen growth and development under salt stress. The results also suggested that anther and pollen wall remodelling/metabolism proteins contribute to the tolerance of rice to salt stress. PMID:22868211

For chromosome duplication of Potato virus Y (PVY)-resistant haploid mutants obtained by the combination treatment of tobacco anther culture and ion beam exposure, pith tissue culture was performed. Thirty-five plants were obtained by pith tissue culture and subsequent acclimatization. Their chromosome number ranged from 22 to 96, and 16 of them had 48 chromosomes. These plants produced fertile seeds by self-pollination. When plants derived from self-pollinated seeds were inoculated with PVY, resistant and susceptible plants were observed. Of the plants derived from the most resistant mutant in the haploid generation, about 50% was resistant to PVY.   

Abstract in portuguese Este estudo visa estabelecer a associação entre o tamanho do botão floral e o estágio de desenvolvimento dos micrósporos em cultivares brasileiras de soja. A distribuição dos estágios de desenvolvimento dos micrósporos foi analisada, nos anos de 1995 e 1997, em botões florais jovens das cultivares Década, IAS5 e RS7. Os dados indicam que, para um dado intervalo de tamanho do botão floral, os micrósporos das diferentes cultivares estavam em distintos estágios (more) de desenvolvimento, com as cultivares RS7 e Década exibindo estágios anteriores àqueles observados na cultivar IAS5. A distribuição dos estágios dos micrósporos entre as dez anteras de um mesmo botão floral foi também analisada nestas cultivares. As dez anteras de um mesmo botão estão claramente distribuídas em diferentes estágios de desenvolvimento. Cautela deve ser tomada ao adotar-se o procedimento padrão em cultura de anteras de utilizar o estágio de desenvolvimento dos micrósporos em uma única antera para representar a totalidade do botão floral. Abstract in english This study was carried out to establish the association between floral bud size and the corresponding microspore developmental stages for Brazilian soybean cultivars. Microspore developmental stage distributions were examined in young buds from cv Década, IAS5 and RS7. The data indicated that for a given bud-size group, the microspores of different cultivars were at different developmental stages, with cv RS7 and Década distributed at the youngest and cv IAS5 at the mos (more) t advanced stages. Microspore stages distribution were also compared among the ten anthers of the same bud of the above cultivars. The ten anthers from a given bud were clearly distributed at different developmental stages. Caution should be exercised when adopting the standard anther culture practice of using the microspore stage of one anther to represent the entire bud.

QTLs associated with ABA content in triticale anthers were analysed using the population of 72 DH lines derived from F1 cross between inbred line ?Saka 3006? and cv. ?Modus?. Using Windows QTL Cartographer and MapQTL 5.0 softwares, composite interval mapping (CIM) and association analysis (Kruskal?Wallis test) for ABA content before and after low temperature treatment inducing androgenic development were conducted. The concentration of ABA in freshly cut tillers varied from 0.8 to 2.2?nmol?g?1 DW. Intensive ABA accumulation (up to 5.3?nmol?g?1 DW) was observed in response to low temperature stress. No correlation between ABA concentration and androgenesis induction efficiency was detected, whereas negative correlation (up to r?=??0.33) was found between concentration of ABA in anthers coll...

Abstract in portuguese Anteras obtidas de botões florais da cultivar IAS-5 de soja foram cultivadas em dois meios de cultura basais (B5 e B5 longo). Análises citológicas das anteras cultivadas in vitro foram realizadas durante os primeiros 20 dias de cultura, a fim de avaliar a viabilidade (por testes de carmim propiônico e FDA) e o estágio de desenvolvimento dos grãos de pólen. As freqüências de grãos de pólen viáveis variaram significativamente entre os tamanhos de botões florais (more) na análise com carmim propiônico. O meio de cultura basal e o tamanho do botão floral não têm um claro efeito nas freqüências de pólens binucleados simétricos e multinucleados. Contagens cromossômicas de micrósporos metafásicos ao longo do período de cultura mostraram nível de ploidia superior ao normal (n=20). Abstract in english Anthers obtained from flowers buds of soybean cultivar IAS-5 were cultured in two basal culture media (B5 and B5 long). Cytological examinations of the in vitro anthers were performed during the first 20 days of culture to assay the viability (by propionic-carmine and fluorescein diacetate tests) and the stage of development of pollen grains. The frequencies of viable pollen grains varied significantly between bud sizes on the propionic-carmine analysis. The basal culture (more) media and bud size had no clear effect on the frequencies of binucleate symmetrical and multinucleate pollen grains. Chromosome counts of metaphasic microspores throughout the culture period showed microspores with higher ploidy level in addition to normal chromosome number (n=20).

The genus Asparagus is very large consisting of around 150 species found as herbaceous perennials, tender woody shrubs and vines. The cultivated species (Asparagus officinalis L., diploid) is a highly prized vegetable, grown in different environments ranging from cool temperate zones to deserts, Mediterranean climates and tropical areas. As a consequence, Asparagus breeders have developed different cultivars that differ for their morpho-agronomic traits, habit and ploidic status (few triploid and tetraploid cultivars are used). Several breeding methods are used for developing cultivars, among which a well developed in vitro anther culture technique produces homozygous clones useful for F1 hybrids constitution. A fluorescent based AFLP (amplified fragment length polymorphism) technique were...

Ploidy levels of in vitro shoots produced through anther culture of asparagus were determined by flow cytometry (FCM) analysis instead of counting the chromosome number of root tips that were difficult to regenerate. Of the 110 anther-derived shoots, there were 83 diploids, 21 tetraploids, 3 octaploids, and 3 mixoploids, but no haploids. When flower buds of different length were used as starting material for anther culture, the richest variation in polyploidy originated from short flower buds (1.0–1.5 mm). This result suggested that polyploids might develop by the fusion of nuclei during pollen-mother cell division before the tetrad stage. We used scanning electron microscopy (SEM) to measure the stomatal length of in vitro and field-grown shoots in order to clarify the differences among polyploids determined by FCM analysis. When the developmental stage of tissue was taken into account, we found statistically significant differences in stomatal length among all polyploids (diploid, tetraploid, and octaploid) for both stems and phyllodes (P < 0.001). Stomata on stems were longer than those on phyllodes, and mature tissue had longer stomata than immature tissue. Morphological observations with SEM found that immature tissue had guard cells set below epidermal cells, and were poorly developed. Field-grown diploids had much shorter stomatal length than in vitro diploids. Male plants grown in the field had significantly longer stomata than female plants in both their immature stems and mature phyllodes (P < 0.001). These results indicated that stomatal length can be a reliable ploidy index by removing any other influential factors such as sex, environment and developmental stage.   

Tree peony (Paeonia suffruticosa) is one of China?s most traditional flowers. It is also one of the most remarkable flowering ornamentals with a rich history of breeding. The speed and success of propagation have always been a major concern for breeders and cultivators. Tissue culture is one of the few effective methods to overcome these inherent difficulties. This article reviews advances made on various aspects of tree peony tissue culture and micropropagation, including the in vitro culture of buds, stem apex, leaves and petioles, sexual embryos and ovules, somatic embryos, anther and callus. Since the vast majority of literature is published in Chinese, this review provides a unique and valuable resource and spring-board from which to initiate or further studies related to in vitro cul...

Our project began on July 1, 1992, with the objective of developing systems that could be used in biotechnological approaches to switchgrass improvement. Within six months after initiation of the project, we had worked out protocols in which plants could be regenerated from callus cultures through both organogenesis and somatic embryogenesis. Documentation for both modes of regeneration was provided in our progress reports and in publications. One thousand regenerated plants were established in the field during the first year. We found that Alamo (lowland type) was much more amenable to in vitro culture, and plants could be regenerated much more easily than from Cave-in-Rock (upland type). During the first three years of the project, we studied the influence of genotype, culture medium components, explant type, etc., on regeneration. As mentioned, we found that the lowland cultivars Alamo and Kanlow were much easier to regenerate than upland cultivars, such as Trailblazer, Blackwell, and Cave-in-Rock. For callus induction, we initially used mature caryopses, young leaf tissue, and portions of seedlings. We were successful in inducing callus and regenerating plants from all explants. Two other systems developed during the 4th to 6th year period of the project included multiple shoot formation initiated from germinated seedlings and regenerable suspension cultures. The latter were initiated from embryogenic calluses produced from in vitro developed inflorescences. An important factor for producing multiple shoots was the presence of thidiazuron in the medium. The shoots could be easily rooted and numerous plantlets produced. The last 3 to 4 years of the project focused on anther and microspore culture experiments to produce haploid plants and on genetic transformation. Although thousands of putative haploid plants were produced from a few anthers, they were very weak and difficult to keep alive. Chromosome counts revealed the gametic number in cells where it was possible to count chromosomes. The isolated microspore culture experiments were not successful.

Anthers of Chinese narcissus (Narcissus tazetta L. var chinesis Roem) were used as explants for callus induction and plant regeneration. About 80% anthers produced callus and 28% of the callus differentiated out bulbs, making a good experiment system of tissue culture of Chinese narcissus for further cellular and gene engineering. The 700 callus were treated by 0.5% colchicin for 5-6 days and then transformed into a MS medium containing 3 mg/L 6-BA to induce differentiation. 90 bulbs were obtained and 55 bulbs among them were checked the chromosome number from their root tips for three times. 29 bulbs (53%, 29/55) still kept triploidy and the most cells of root tips contained 30 chromosomes. 22 bulbs (40%, 22/55) displayed aneuploidy and the most cells of its root tips contained 10-50 chromosomes. 4 bulbs displayed hexaploidy and contained 60 chromosomes. After three months growing, the cells of root tips containing aneuploidy chromosomes disappeared, and the bulbs became triploidy. The chromosomes of 4 hexaploidy bulbs did not changed during three checks. The origin and disappearance of aneuploidy cells of Chinese narcissus after treated by colchicin were discussed. PMID:17674781

The purpose of this microscopic histochemical study was to establish the developmental sequence leading to sterility in a cytoplasmic male sterile Helianthus petiolaris line. Histochemical studies showed that CMS anthers, in contrast to fertile anthers, lacked carbohydrate storage at sporogenous stage. CMS sporogenous cells were thick-walled and ascorbic acid-rich. Subsequent developmental pattern was not uniform in CMS anthers. The variations were found in different anthers and in different locules of the same anther. Two categories of CMS anthers lacked callose deposition and showed premature disintegration of tapetal cell layer, which lead to precocious formation of periplasmodium. In one type, CMS anthers contained naked microspores. In another type, coenocytic microspores were formed. The microspores lacked reserve metabolites. In the third category of CMS anthers normal callose deposition and tetrad formation were observed. In this category callose deposition and hypertrophied tapetal cell layer persisted until mature stage of the anther. Results in all 3 categories of CMS anthers link tapetal abnormality to male sterility.   

Cultivated and wild potato species synthesize a wide variety of steroidal glycoalkaloids (GA) that may affect either human health or biotic stress resistance. Therefore, GA composition must be a major criterion in the evaluation of breeding products when species genomes are merged and/or manipulated. This work reports the results of GA analysis performed on unique haploid (2n=2x=24) plants obtained from tetraploid (2n=4x=48) Solanum bulbocastanum-S. tuberosum hybrids through in vitro anther culture. Glycoalkaloids were extracted from tubers and analyzed by HPLC. Haploids generally showed the occurrence of parental GA. However, in several cases loss of parental GA and gain of new GA lacking in the parents was observed. It may be hypothesized that new GA profiles of our haploids is the resul...

Abstract in english Brugmansia candida (Solanaceae) is a native tree distributed across South-American and produces the pharmacologically- important group of tropane alkaloids including scopolamine. This biocompound is synthesised from hyoscyamine by action of Hyoscyamine 6-? hydroxylase (H6H, EC 1.14.11.11) at the end of the tropane alkaloid pathway. Here are reported the tissue and organ-specific expression of h6hmRNA by RT-PCR analyses and the isolation, cloning and sequencing of the (more) cDNA obtained from B. candida anthers and hairy root transformed cultures. Bioinformatic analysis of the nucleotide sequence revealed an uninterrupted ORF of 1038 bp and the predicted aminoacid sequence could be 344 aminoacid long. A database search showed that this sequence has high homology (97% identity) to Hyoscyamus niger H6H protein (Genbank accession number AAA33387.1).

The effect of anther-derived substances on pollen function was studied using pollen produced by in vitro culture of immature pollen of tobacco (Nicotiana tabacum L.) and petunia (Petunia hybrida). Addition of conditioned medium consisting of diffusates from in situ matured pollen strongly increased pollen germination frequency and pollen tube growth, as well as seed set after in situ pollination. Thin-layer chromatography and depletion of phenolic substances by Dowex treatment indicated that flavonols are present in the diffusate and may be the active compounds. When added to the germination medium, flavonols (quercetin, kaempferol, myricetin) but not other flavonoids strongly promoted pollen germination frequency and pollen tube growth in vitro. The best results were obtained at very low concentrations of the flavonols (0.15-1.5 mum), indicating a signaling function. The same compounds were also effective when added during pollen development in vitro. PMID:16653074

A protocol for raising highly frost-resistant winter wheat Triticum aestivum varieties has been developed. It is based on the transfer of the frost resistance trait from a wild wheat relative, intermediate wheatgrass (Agropyron glaucum). The protocol includes raising wheatgrass double haploid via anther culture, the selection of wheat genotypes that are able to produce green haploids in vitro, a proximate analysis of frost resistance, remote crosses of frost-resistant wheatgrass genotypes to wheat, and a new leaf-nurse method for transferring frost resistance from wheatgrass to wheat. A large collection of A. glaucum genotypes has been derived from the original material collected in Eastern Kazakhstan, at an elevated site with little snow. The climate of the site implies that the accession...

Reproductive studies were carried out on Brazilian accessions of Cephaelis ipecacuanha, which is considered a threatened species. Meiotic behavior was studied using squashing technique with 1% acetic carmine and floral buds and anther were measured for the following male gamete development stages: meiosis I, meiosis II, tetrads, microspores and pollen. Bud and anther length showed significant differences (P<0.01) among the stages of pollen development. There was high significant correlation between bud and anther length by Pearson correlation. Tukey test results indicated that each analyzed stage could be associated to bud length mean but it was not found the same association for anther length.   

Abstract in portuguese Anteras de duas cultivares de soja foram cultivadas em meio de cultura basal B5 longo gelificado com agarose ou Phytagel®. Análises citológicas das anteras foram conduzidas durante os primeiros 45 dias de cultura para avaliar a viabilidade e o estágio de desenvolvimento dos micrósporos. A freqüência de formação de calos foi analisada após 45 dias do início da cultura. A análise da variância da viabilidade do micrósporo mostrou interações significativas de (more) Cultivar X Agente Gelificante X Dias de Cultura. As freqüências de grãos de pólen viáveis diminuíram significativamente com o tempo de cultura, dentro de cada cultivar e agente gelificante testado. A interação Dia X Cultivar foi significante para as freqüências de grãos de pólen binucleados simétricos e estruturas multinucleados/multicelulares. O efeito do agente gelificante na freqüência de grãos de pólen binucleados simétricos e estruturas multinucleados/multicelulares não foi significante. Com relação às freqüências de calos e estruturas embriogênicas formadas, houve diferença significativa entre cultivares (IAS5= 14.8% e BRS 133=6.6%). O agente gelificante não mostrou efeito em tais freqüências. Abstract in english Anthers of two soybean cultivars were cultured in B5 long basal culture media gelled with agarose or PhytagelTM. Cytological examinations of the anthers were carried out during the first 45 days of culture to assay the viability and developmental stage of microspores. Frequency of callus formation was recorded at 45 days of culture. The analysis of variance of the microspore viability assay showed significant Cultivar X Gelling Agent X Day of Culture interactions. The fre (more) quencies of viable microspores decreased significantly with time of culture, within each cultivar and gelling agent tested. The interaction Day X Cultivar was significant for the frequencies of binucleate symmetrical grains and multinucleate/multicellular structures. The effect of gelling agents on the frequency of binucleate symmetrical pollens grains and multinucleate/multicellular structures was not significant. About the frequencies of calli and embryogenic calli formed, a significant difference was detected between the cultivars (IAS5= 14.8% and BRS 133=6.6%). Gelling agents showed no effect over these frequencies.

Abstract in portuguese Sementes de gergelin (Sesamum indicum L.) foram cultivadas in vitro em meio de cultura de Wetherall contendo 0,5 mg/l de 2,4-D e em seguida transferidas para meio de Murashige e Skoog (MS) contendo 0,1 mg/l de 2,4-D e 100 mg/l de inositol. Ambos, 2,4-D e inositol mostraram-se ser necessários para o desenvolvimento de calos a partir de sementes, do mesmo modo que para o contínuo crescimento dos meios em cultura. Foram também obtidos calos de explantes de anteras, cotiledones e de hipocotilo de Sesamum utilizando-se o meio MS com a ocorrência de estruturas globulares. Abstract in english Continuously growing Sesamum hypocotyl callus cultures were successfully initiated from hypocotyl tissues of seeds cultured on Wetherall's Medium containing 0.5 mg/l 2,4-D and subcultured on Murashige and Skoog (MS) medium containing 0.1 mg/1 2,4-D and 100 mg/l inositol. Both 2,4-D and inositol appear to be essential for maintenance of continous growth. Callus cultures were likewise established from explants of anthers, cotyledon, and hypocotyl on the MS medium with the occurrence of arrested globular structures in some cultures.

Abstract in portuguese Realizaram-se dois experimentos, o primeiro visando conhecer a capacidade androgênica de variedades de trigo e identificar as melhores condições para alcançar aquele objetivo, em termos de meios de cultura e microclima. No segundo, estudou-se maior número de variedades. No primeiro experimento, foram testadas as seguintes: PF 853031 e IAC 24, a primeira, usada como padrão; dois meios de cultura básicos, batata-2 e N6; duas auxinas, IAA e 2,4-D, nas doses de 3, 9 e (more) 27 ìmol/L, e cinetina, nas doses de 2, 6, 12 e 24 ìmol/L. As anteras plaqueadas foram acondicionadas em sala fotoperiódica: (a) com 14 h de luz, 60 ìmol.m-2.s-1 (3.200 lux), à temperatura de 25°C, e (b) submetidas a pré-tratamento por quatro dias a 6°C e, em seguida, retornadas à condição a. Notou-se que a melhor combinação de tratamentos para a variedade IAC 24 foi o meio batata-2, 2,4-D a 27 .ìmol/L e cinetina 2 a 6 ìmol/L, por promover maior indução de estruturas androgênicas. Já para a variedade PF 853031, a mesma combinação de tratamentos, exceto 2,4-D a 9 µmol/L, promoveu também maior indução de estruturas androgênicas. Por outro lado, o pré-tratamento de quatro dias de frio causou maior indução de estruturas androgênicas em ambas as variedades. No segundo experimento, foram estudadas cinco variedades: PF 853048, usada como padrão, IAC 21, BH 1146, IAC 60 e Anahuac; dois meios de cultura básicos: batata-2 e N6; duas combinações hormonais: CH1 (10 µmol/L IAA e 30 ìmol/L de cinetina), e CH2 (10 ìmol/L de 2,4-D e 3,0 ìmol/L de cinetina). Observou-se maior indução de estruturas androgênicas para a 'PF 853048', seguida da 'Anahuac'. Todavia, somente foram obtidas plantas para a primeira. Essa constatação sugere que a capacidade androgênica seja controlada geneticamente no trigo. Na combinação hormonal 10 ìmol/L de 2,4-D e 3 ìmol/L de cinetina houve maior formação de estruturas androgênicas. Transplantaram-se as plantas regeneradas para vasos contendo solo, alocando-as em casa de vegetação para aclimatação, crescimento e florescimento. As plantas mostraram-se inférteis, mas produziram sementes após a duplicação pela técnica da colchicina. Abstract in english Experiments were carried out to establish conditions and to access varietal effects for wheat androgenesis. The varieties PF 853031, a standard known for androgenic capacity, and the IAC 24 of high agronomic value, were tested in two culture mediums, potato-2 and N6, including two auxins, IAA and 2.4-D, at 3, 9 and 27 ìmol/L, and kinetin, at 2, 6, 12 and 24 µmol/L. The experiment was installed in factorial design with six blocks and 20 anthers per plate (a block). (more) About 5,760 anthers were plated per variety. The experiment was left in a photoperiodic room with (a) 14 h of cool fluorescent light, 60 ìmol.m-2.s-1 (3,200 lux), at temperature of 25°C, and (b) pretreated in dark at 6°C for four days, then backed to a. The variety IAC 24 showed higher androgenic capacity than the PF 853031 variety, in medium potato-2, 2.4-D at 27 ìmol/L and kinetin from 2 to 6 µmol/L. Cold treatment for four days was beneficial in improving androgenesis to both varieties. Five other varieties were studied, PF 853048, as a androgenic control, IAC 21, BH 1146, IAC 60 and Anahuac; two culture mediums, potato-2 and N6, along with two hormonal combinations, CH1 (IAA 10 µmol/L and kinetin 30 µmol/L) and CH2 (2.4-D 10µmol/L and kin 3 .mol/L) in the condition as (b) above. The standard variety PF 853048 ranked first and Anahuac, the second in terms of androgenic response. However, plants were only obtained from PF 853048. It is likely that the androgenic effect is under genotype control in wheat. The hormonal combination 10 µmol/L of 2.4-D and 3 µmol/L of kinetin were observed to induce higher androgenic response. Regenerated plants were potted and left to flower. All of them showed to be infertile, but produced seeds after colchicine treatment.

The effect in vitro of washed and crushed anther/pollen and stigma extracts from six sorghum lines with different levels of ergot susceptibility on ergot conidial germination was determined. Conidia plated on water agar amended with either anther/pollen or stigma extracts generally stimulated highe...

In this study, polysaccharide and RNA contents of anthers were investigated on different phases of sporogenesis by using light microscopy techniques from histological and cytological point of view in Leucojum aestivum. Paraffin and semi-thin sections of anthers were stained with toluidine blue and PAS. Anthers were tetrasporangiate. The wall of the anther consists of an epidermis, endothecium, middle layer and glandular tapetum. During one nucleated microspore and mature pollen phase microspores and tapetum cells began to degenerate and they were become very rich of RNA in L. aestivum. And also RNA content was increased in endothecium and middle layer cells except the epidermis cells of anther wall. An increase in RNA content indicates cell activation. Polysaccharides were not seen in youn...

The Role of Culture in Elder Abuse Cultural values, beliefs, and traditions significantly affect family life. They dictate family members' roles and responsibilities toward one another, how family ...

The authors present an approach for conducting and interpreting results of newly established plant cell culture in toxicity studies. xtended culturing produces uniform suspension and facilities sampling. rimary (new) cultures are more representative of all responses of their plan...

Anther and pollen development in staminate and pistillate flowers of dioecious Melicoccus lepidopetalus (Sapindaceae) were examined by light and electron microscopy. Young anthers are similar in both types of flowers; they consist of epidermis, endothecium, two to four middle layers and a secretory tapetum. The microspore tetrads are tetrahedral. The mature anther in staminate flowers presents compressed epidermal cells and endothecium cells with fibrillar thickenings. A single locule is formed in the theca by dissolution of the septum and pollen grains are shed at two-celled stage. The mature anthers of pistillate flowers differ anatomically from those of staminate flowers. The epidermis is not compressed, the endothecium does not develop fibrillar thickenings, middle layers and tapetum a...

Dyckia pseudococcinea L.B.Sm. is endemic to the restingas of Marica, State of Rio de Janeiro, in southeastern Brazil. However, since this area is under intense ecological stress, D. pseudococcinea is considered an endangered plant species. An important step toward the conservation of this species is establishing the developmental stage most favorable to in vitro propagation. Accordingly, the present study analyzed the androecium to establish the developmental stages of anthers, emphasizing anther wall development, androsporogenesis and androgametogenesis. The relationships between the size of flowers and anthers were also studied. Anther wall development follows the basic-type, while the tapetum, which is originated from the subepidermal layer (inner secondary parietal layer 2 and archespo...

  Beta-1,3-glucanases are referred to as pathogenesis-related proteins and they are also involved in several developmental processes. We isolated a cDNA for ?-1,3-glucanase from rice anther and named it Oryza sativa glucanase 1 (Osg1). Phylogenetic analysis showed that Osg1 belonged to monocotyledonous endo-?-1,3-glucanase subgroup A. RT-PCR analysis revealed that Osg1 transcripts were present in leaves, roots, and anthers.   

The maize (Zea mays) late pollen gene ZmMADS2 belongs to the MIKC type of MADS box transcription factor genes. Here, we report that ZmMADS2, which forms a homodimer in yeast (Saccharomyces cerevisiae), is required for anther dehiscence and pollen maturation. Development of anthers and pollen was arrested at 1 d before dehiscence in transgenic plants expressing the ZmMADS2-cDNA in antisense orientation. Temporal and spatial expression analyses showed high amounts of ZmMADS2 transcripts in endothecium and connective tissues of the anther at 1 d before dehiscence and in mature pollen after dehiscence. Transient transformation of maize and tobacco (Nicotiana tabacum) pollen with the luciferase reporter gene under the control of different ZmMADS2 promoter deletion constructs demonstrated the functionality and tissue specificity of the promoter. Transgenic maize plants expressing a ZmMADS2-green fluorescent protein fusion protein under control of the ZmMADS2 promoter were used to monitor protein localization during anther maturation and pollen tube growth. High amounts of the fusion protein accumulate in degenerating nuclei of endothecial and connective cells of the anther. A possible function of ZmMADS2 during anther dehiscence and pollen maturation and during pollen tube growth is discussed. PMID:15001699

Cultivated and wild potato species synthesize a wide variety of steroidal glycoalkaloids (GA) that may affect either human health or biotic stress resistance. Therefore, GA composition must be a major criterion in the evaluation of breeding products when species genomes are merged and/or manipulated. This work reports the results of GA analysis performed on unique haploid (2n=2x=24) plants obtained from tetraploid (2n=4x=48) Solanum bulbocastanum-S. tuberosum hybrids through in vitro anther culture. Glycoalkaloids were extracted from tubers and analyzed by HPLC. Haploids generally showed the occurrence of parental GA. However, in several cases loss of parental GA and gain of new GA lacking in the parents was observed. It may be hypothesized that new GA profiles of our haploids is the result of either genetic recombination or combinatorial biochemistry events. To highlight differences between haploids and parents, soluble proteins and antioxidant activities were also determined. Both were always higher in haploids compared to their parents. The nature of the newly formed GAs will be further investigated, because they may represent new metabolites that can be used against pest and diseases, or are useful for human health. PMID:20730954

We present data on the morphological, cytological, biochemical and genetic characteristics of tomato regenerants obtained through anther culture. As a result of induced androgenesis, more than 6,000 rooted regenerants were developed that differed both from the donor plants and among each other with respect to habitus and leaf, flower and inflorescence morphology. Cytological analysis revealed a great variability in chromosome number in the cells of the regenerated plants. While most of the regenerants were mixoploid, the majority of the cells had a haploid chromosome number. R(1) and R(2) progenies were tested for their resistance to Clavibacter michiganense subsp. michiganense ( Cmm 7). Some of the regenerants were resistant to the pathogen. A biochemical analysis of fruit from R(3) and R(4) plants showed a higher content of dry matter, sugars and vitamin C in the regenerant plants obtained from the hybrids than in those from the cultivars and control plants. The values of the parameters of hybrid regenerants grown in the greenhouse were about 1.5-fold higher than those of the hybrid regenerants grown in the field, and this trend is clearly expressed in all of the hybrid regenerants. The results obtained suggest that induced androgenesis and gametoclonal variation may be used as an additional tool to create a large range of new forms. The application of the latter in breeding programs would accelerate the development of tomato lines and varieties that would be more productive, disease-resistant, highly nutritive and flavour-acceptable. PMID:14551733

Transposon display (TD) is a powerful technique to identify the integration site of transposons in gene tagging as a functional genomic tool for elucidating gene function. Although active endogenous DNA transposons have been used extensively for gene tagging in maize, only two active endogenous DNA transposons in rice have been identified, the 0.43-kb element mPing of the MITE family and the 0.6-kb nDart element of the hAT family. The nDart transposition was shown to be induced by crossing with a line containing its autonomous element aDart and stabilized by segregating aDart under natural growth conditions, while mPing-related elements were shown to transpose in cultured cells, plants regenerated from an anther culture, and ?-ray-irradiated plants. No somaclonal variation should occur in nDart-promoted gene tagging because no tissue culture was involved in nDart activation. As an initial step to develop an effective tagging system using nDart in rice, we tried to visualize GC-rich nDart-related elements comprising 18 nDart-related sequences of 0.6-kb and 63 nDart-related elements longer than 2 kb in Nipponbare by TD. Comparing the observed bands in TD with the anticipated virtual bands of the nDart-related elements based upon the available rice genome sequence, we have improved our TD protocol by optimizing the PCR amplification conditions and are able to visualize approximately 87% of the anticipated bands produced from the nDart-related elements. To compare the visualization efficiency of these nDart-related elements with that of 50 mPing elements and a unique Ping sequence in Nipponbare, we also tried to visualize the mPing-related elements; all mPing-related elements are easily visualized. Based on these results, we discuss the parameters affecting the visualization efficiencies of these rice DNA transposons. We also discuss the utilization of nDart elements in gene tagging for functional genomics in rice.   

As is the case with other organisms, plants respond to genotoxic stresses by expressing DNA repair genes upon DNA damage. To uncover the mechanisms involved in regulated expression of the DNA damage-responsive gene, we investigated the tissue specific expression and DNA damage-responsiveness of the Arabidopsis RAD51 (AtRAD51) gene promoter in Arabidopsis and tobacco. Transgenic Arabidopsis and tobacco plants harboring AtRAD51 promoter- ?-glucuronidase (GUS) were used to study the detailed expression pattern of the AtRAD51 gene. A histochemical GUS assay of bleomycin- or UV-treated plants showed that the AtRAD51 promoter in young tissues is actively expressed particularly in meristematic cells of the root and shoot apex of seedlings. In the absence of genotoxic stress, GUS activities were detected only at very low levels in these same organs. In mature plants, the AtRAD51 promoter is mainly expressed in flower bud, sepal, stigma, later anther, pedicel when treating with DNA damaging agent. The expression patterns of reporter assays were consistent with the AtRAD51 mRNA accumulation pattern. These results suggest that the regulated expression of the AtRAD51 gene is controlled mainly at the level of transcription directed primarily by the promoter function of the gene.   

Brassinosteroids (BRs) are plant hormones that regulate growth and development. They share structural similarities with animal steroids, which are decisive factors of sex determination. BRs are known to regulate morphogenesis and environmental stress responses, but their involvement in sex determination in plants has been only speculative. We show that BRs control sex determination in maize revealed through characterization of the classical dwarf mutant nana plant1 (na1), which also feminizes male flowers. na1 plants carry a loss-of-function mutation in a DET2 homolog--a gene in the BR biosynthetic pathway. The mutant accumulates the DET2-specific substrate (24R)-24-methylcholest-4-en-3-one with a concomitant decrease of downstream BR metabolites. Treatment of wild-type maize plants with BR biosynthesis inhibitors completely mimicked both dwarf and tasselseed phenotypes of na1 mutants. Tissue-specific na1 expression in anthers throughout their development supports the hypothesis that BRs promote masculinity of the male inflorescence. These findings suggest that, in the monoecious plant maize, BRs have been coopted to perform a sex determination function not found in plants with bisexual flowers. PMID:22106275

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Anther structure and pollen ontogeny of six species of Hosta Tratt. was studied to provide additional data for its classification. Microscopic observation found that their male gametogenesis has the following structural characteristics: (1) the four microsporangia are linearly arranged on introrse side of anther connective tissue; (2) mature anther wall is composed of an epidermis, endothecium, middle layer(s), and tapetum; (3) microsporogenesis is successive; (4) mature pollen grains are three-celled at anthesis, with a vegetative cell filled with large amounts of starch grains; (5) pollen ontogeny is obviously asynchronous; (6) each tepal has about 9?13 vascular traces in cross-section view. The above observations suggest that genus H. Tratt. should be classified as family Hostaceae, in ...

It is not well documented which floral parts may trigger the onset of tepal or petal senescence and or flower bud abscission. Asiatic hybrid (Liliumxelegans Thunb.) 'Red Carpet' lily flowers were selected as a model to study this relationship because the various floral organs of a lily flower can be easily dissected and collected for physiological studies. We hypothesized that anther is the organ that triggers flower bud abscission. Ethanol-soluble sucrose, fructose, and glucose were analyzed in the tepal, anther, filament, and pistil. The analysis was conducted on flower buds weighing 1.5g (small buds) and 3.9g (large buds). The sucrose content in the anthers of small buds remained high when cut flowers were held at 21^oC for 5d, suggesting that sucrose is not hydrolyzed into fructose and...

The most chilling-sensitive stage of rice has been found to be at the onset of microspore release. The microsporocytes produce a wall of callose between the primary cell wall and the plasma membrane, and it has been shown that precise regulation of callose synthesis and degradation in anther is essential for fertile pollen formation. In this study, genes for 10 callose synthases in the rice genome were fully annotated and phylogenetically analyzed. Expression analysis of these genes showed that OsGSL5, an ortholog of microsporogenesis-related AtGSL2, was specifically expressed in anthers, and was notably downregulated by cooling treatment. Gene expression profiles of Rho-type small GTP-binding proteins in rice anther were also analyzed. The mechanisms of callose synthesis in rice pollen formation and its relationships with cool tolerance are discussed.   

Bioluminescence, mRNA levels, and toluene degradation rates in Pseudomonas putida TVA8 were measured as a function of various concentrations of toluene and trichloroethylene (TCE). TVA8 showed an increasing bioluminescence response to increasing TCE and toluene concentrations. Compared to uninduced TVA8 cultures, todC1 mRNA levels increased 11-fold for TCE-treated cultures and 13-fold for toluene-treated cultures. Compared to uninduced P. putida F1 cultures, todC1 mRNA levels increased 4,4-fold for TCE-induced cultures and 4.9-fold for toluene-induced cultures. Initial toluene degradation rates were linearly correlated with specific bioluminescence in TVA8 cultures.

Agrobacterium tumefaciens-mediated genetic transformation and the regeneration of transgenic plants was achieved in Hevea brasiliensis. Immature anther-derived calli were used to develop transgenic plants. These calli were co-cultured with A. tumefaciens harboring a plasmid vector containing the H. brasiliensis superoxide dismutase gene (HbSOD) under the control of the CaMV 35S promoter. The beta-glucuronidase gene (uidA) was used for screening and the neomycin phosphotransferase gene (nptII) was used for selection of the transformed calli. Factors such as co-cultivation time, co-cultivation media and kanamycin concentration were assessed to establish optimal conditions for the selection of transformed callus lines. Transformed calli surviving on medium containing 300 mg l(-1) kanamycin showed a strong GUS-positive reaction. Somatic embryos were then regenerated from these transgenic calli on MS2 medium containing 2.0 mg l(-1) spermine and 0.1 mg l(-1) abscisic acid. Mature embryos were germinated and developed into plantlets on MS4 medium supplemented with 0.2 mg l(-1) gibberellic acid, 0.2 mg l(-1) kinetin (KIN) and 0.1 mg l(-1) indole-3-acetic acid. A transformation frequency of 4% was achieved. The morphology of the transgenic plants was similar to that of untransformed plants. Histochemical GUS assay revealed the expression of the uidA gene in embryos as well as leaves of transgenic plants. The presence of the uidA, nptII and HbSOD genes in the Hevea genome was confirmed by polymerase chain reaction amplification and genomic Southern blot hybridization analyses. PMID:14551734

Screening of an anther cDNA expression library resulted in the isolation of two almost identical cDNA clones, termed mipA and mipB, showing homology with sequences encoding transmembrane channel proteins from the MIP family. Both clones were expressed in several tissues, but not in pollen. MipA was preferentially expressed in the surrounding sporophytic tissues of stamens. Anthers subjected to drought were induced to accumulate even more mip transcripts, which was entirely due to higher mipA gene expression. On basis of isolation procedures, sequence homology and drought inducibility of mipA we conclude that the encoded proteins probably are constituents of the pollen coat and are aquaporins. PMID:9177322

The embryological features of microsporangiate wall formation, microsporogenesis and male gametogenesis in Aster subulatus, Kalimeris indica, Heteropappus arenarius and Erigeron annuus are described for the first time. Although external morphology might suggest that these four species differ from each another, we found no distinctly different embryological features. They all share similar characteristics, such as a tetrasporangiate anther, coexisting amoeboid and glandular tapeta, simultaneous cytokinesis (except for 12% occurrence of successive cytokinesis in E. annuus), 3-celled mature pollen grains and endothecium of anther wall that does not develop fibrous thickenings. Based on the embryological data, we suggest that Kalimeris and Heteropappus may be included within Aster, and that Er...

BACKGROUND:: Culture change initiatives propose to improve care by addressing the lack of managerial supports and prevalent stressful work environments in the industry; however, little is known about how culture change facilities differ from facilities in the industry that have not chosen to affiliate with the resident-centered care movements. PURPOSE:: The aim of this study was to evaluate representation of organizational culture values within a random sample of U.S. nursing home facilities using the competing values framework and to determine whether organizational values are related to membership in resident-centered culture change initiatives. DESIGN AND METHODS:: We collected reports of cultural values using a well-established competing values framework instrument in a random survey of facility administrators and directors of nursing within all states. We received responses from 57% of the facilities that were mailed the survey. Directors of nursing and administrators did not differ significantly in their reports of culture and facility measures combined their responses. FINDINGS:: Nursing facilities favored market-focused cultural values on average, and developmental values, key to innovation, were the least common across all nursing homes. Approximately 17% of the facilities reported that all cultural values were strong within their facilities. Only high developmental cultural values were linked to participation in culture change initiatives. Culture change facilities were not different from non-culture change facilities in the promotion of employee focus as organizational culture, as emphasized in group culture values. Likewise, culture change facilities were also not more likely to have hierarchical or market foci than non-culture change facilities. PRACTICE IMPLICATIONS:: Our results counter the argument that culture change facilities have a stronger internal employee focus than facilities more generally but do show that culture change facilities report stronger developmental cultures than non-culture change facilities, which indicates a potential to be innovative in their strategies. Facilities are culturally ready to become resident centered and may face other barriers to adopting these practices. PMID:22936002

This second part of Mythologising Culture examines the responses of Aboriginal people in western Sydney to the valorizing of Aboriginality by the Australian nation. As Aboriginal culture has become the object of restitution, regret and reconciliation, Aboriginal people are being called upon to represent and produce Culture in iconic forms such as painting, dancing and other performances and representations. These activities, often depicted as remedial, are seldom initiated and controlled by Aboriginal people, but rather disturb cultural adjustments and established social relations. I discuss a range of responses elicited by these post-colonial activities, from enthusiastic embrace to the rejection of state sponsored culture. Further, I consider the structural implications of suburban Abori...

1. gamma-Aminobutyric acid (GABA) responses were recorded from rat superior cervical ganglia (SCG) in culture using the whole cell recording technique. 2. Zinc (50-300 microM) reversibly antagonized the GABA response in embryonic and young post-natal neurones, while neurones cultured from adult anim...

The development of anther and pollen is important for male reproduction, and this process is coordinately regulated by many external and internal cues. In this study, we systematically examined the male reproductive phenotypes of a series of brassinosteroid biosynthetic and signaling mutants and fou...

In flowering plants, the male gametophyte, the pollen, develops in the anther. Complex patterns of gene expression in both the gametophytic and sporophytic tissues of the anther regulate this process. The gene expression profiles of the microspore/pollen and the sporophytic tapetum are of particular interest. In this study, a microarray technique combined with laser microdissection (44K LM-microarray) was developed and used to characterize separately the transcriptomes of the microspore/pollen and tapetum in rice. Expression profiles of 11 known tapetum specific-genes were consistent with previous reports. Based on their spatial and temporal expression patterns, 140 genes which had been previously defined as anther specific were further classified as male gametophyte specific (71 genes, 51%), tapetum-specific (seven genes, 5%) or expressed in both male gametophyte and tapetum (62 genes, 44%). These results indicate that the 44K LM-microarray is a reliable tool to analyze the gene expression profiles of two important cell types in the anther, the microspore/pollen and tapetum.

3-Di-p-coumaroylsophoroside-5-malonylglucoside and its demalonyl derivative were isolated from blue petals of Veronica persica Poiret. Blue, violet and purple cells coexist in the petal. These colors might be due to the varying pH of the vacuole between 5 and 7 unit. Only the demalonylated pigment was detected in the blue anthers.   

Abstract in portuguese Flores de três clones de Hevea brasiliensis, RRIM 527, RRIM 600 e GT 1, foram analisadas, sob lupa e microscopia eletrônica de varredura, a fim de se observar as taxas de ocorrência de hermafroditismo. Os resultados mostraram um total de 71,49% de flores hermafroditas, sendo que destas 29,83% apresentaram anteras residuais, não completamente desenvolvidas. As análises ao microscópio de varredura não mostraram diferença ao nível de epiderme de anteras em flores ma (more) sculinas e hermafroditas de RRIM 527 e RRIM 600. No clone GT 1 (macho estéril) a epiderme das anteras mostrou-se frouxa já no início do desenvolvimento floral e completamente enrugada ao final da maturação, demonstrando que as anteras estão vazias neste estádio. Abstract in english Flowers of three Hevea brasiliensis clones, RRIM 527, RRIM 600 and GT 1, were analyzed under stereomicroscope and scanning electron microscope, aiming to observe hermaphroditism rates. Results showed 71.49% hermaphrodite flowers, 29.83% of which exhibited incompletely developed, residual anthers. The scanning electron microscope analysis did not detect differences in anther epidermis of male and bisexual flowers of RRIM 600 and RRIM 527. In GT 1 clone (sterile male), the (more) anther epidermis was already weak at the beginning of floral development and completely wrinkled at the end of maturation. Consequently, the anthers were empty by this stage.

The occurrence of polytenic nuclei in the anther tapetum of field grown variety K 5269 of a bean species, Vigna unguiculata has been recorded. There are pronounced polytene nuclei with chromocenters unassociated with chromatin bundles. In comparison to polytenic nuclei of other plants they were relatively smaller with a small nucleolus. These features of polytenic nuclei suggest low level of endoreduplication in Vigna unguiculata.   

Cultural evolutionary theory is an interdisciplinary field in which human culture is viewed as a Darwinian process of variation, competition, and inheritance, and the tools, methods, and theories developed by evolutionary biologists to study genetic evolution are adapted to study cultural change. It is argued here that an integration of the theories and findings of mainstream social psychology and of cultural evolutionary theory can be mutually beneficial. Social psychology provides cultural evolution with a set of empirically verified microevolutionary cultural processes, such as conformity, model-based biases, and content biases, that are responsible for specific patterns of cultural change. Cultural evolutionary theory provides social psychology with ultimate explanations for, and an understanding of the population-level consequences of, many social psychological phenomena, such as social learning, conformity, social comparison, and intergroup processes, as well as linking social psychology with other social science disciplines such as cultural anthropology, archaeology, and sociology. (Contains 4 footnotes, 1 table, and 2 figures.)

Previously, it was shown that ?-ketoacyl-CoA synthase CER6 is necessary for biosynthesis of very-long-chain fatty acids (VLCFAs) with chain lengths beyond C28 in Solanum lycopersicum fruits and C26 in Arabidopsis thaliana leaves and the pollen coat. CER6 loss-off-function in Arabidopsis resulted in conditional male sterility since pollen coat lipids are responsible for contact-mediated pollen hydration. In tomato, on the contrary, the pollen hydration does not rely on the pollen coat lipids. Nevertheless, mutation in SlCER6 impairs the fertility and floral morphology. Herein, the contribution of SlCER6 to the sexual reproduction and flower development in tomato was addressed. Cytological analysis and cross-pollination experiments revealed that the slcer6 mutant has male sterility caused by (i) hampered pollen dispersal and (ii) abnormal tapetum development. SlCER6 loss-of-function provokes a decrease of n- and iso-alkanes with chain lengths ? C27 and of anteiso-alkanes with chain lengths ? C28 in flower cuticular waxes, but has no impact on the flower cuticle ultrastructure and the cutin content. Expression analysis confirmed high transcription levels of SlCER6 in the anther and the petal, preferentially in sites subject to the epidermal fusion. Hence, wax deficiency was proposed to be the primary reason for the flower fusion phenomena in tomato. The SlCER6 substrate specificity was revisited. It might be involved in elongation of not only linear but also branched VLCFAs, leading to production of the corresponding alkanes. SlCER6 implements a function in the sexual reproduction of tomato which is different from the one in Arabidopsis: SlCER6 is essential for regulation of timely tapetum degradation and, consequently, microgametogenesis. PMID:23144186

Objective: This second paper follows an exploration of the nature of blame and addresses the balance between a `?blame-free'? health culture and appropriate accountability. This paper aims to define and describe accountability as a key component of clinical governance and a responsive, fair and transparent health culture. Methods: The literature is examined and the concept of a fair and transparent health culture is explored. The case vignette in Part 1 is used to illustrate a particular issue of accountability. Results: The place of accountability in relation to clinicians and health organisations is elucidated. Conclusions: The necessary conditions for an accountable, responsive, fair and transparent health culture are proposed.

Psychiatrists and anthropologists have taken distinct analytic approaches when confronted with differences between emic and etic models for distress: psychiatrists have translated folk models into diagnostic categories whereas anthropologists have emphasized culture-specific meanings of illness. The rift between psychiatric and anthropological research keeps "individual disease" and "culture" disconnected and thus hinders the study of interrelationships between mental health and culture. In this article we bridge psychiatric and anthropological approaches by using cultural models to explore the experience of nerves among 27 older primary care patients from Baltimore, Maryland. We suggest that cultural models of distress arise in response to personal experiences, and in turn, shape those ex...

To study calcium imaging data of cell populations that have various response patterns in peak amplitude and frequency of calcium oscillation in response to stimulation, comprehensive characterization based on statistical analysis of each response is important. In cultures of cells that are flat and in contact with each other, it is difficult to distinguish individual cells in calcium imaging data. We have developed a novel method to determine areas corresponding to individual cells in calcium imaging data. Rat neonatal cerebral astrocytes were filled with the calcium indicator Fura2, stained with acridine orange, and illuminated with UV light. The cell nuclei were clearly visualized. In addition, the images of these nuclei were useful for analyzing concentration-dependent alteration of calcium oscillation of cultured astrocytes in response to glutamate. This novel method may be useful for studying factors affecting calcium response patterns of cultured cell populations, including culture conditions, stimulus paradigms, and synthetic compounds.   

Phytohemagglutinin response of peripheral blood lymphocytes (PBL) of sheep was studied. Assessment of proliferative response was performed by determination of nuclear volumes and cell counts in cultures from 14 sheep and by incorporation of tritiated thymidine in cultures in four additional sheep. PBL of sheep were found to transform and proliferate with PHA similarly to human peripheral blood lymphocytes with minor differences. Quantitation of the proliferative response by determining the cell count and nuclear volumes provided more information on cell kinetics in culture than the commonly used isotope-labeled thymidine incorporation method.

Abstract in spanish El objetivo de esta investigación embriológica,la primera realizada en el género, fue tipificar el proceso de microsporogénesis en Hymenocallis caribaea. La meiosis se desarrolla cuando los botones florales aún se encuentran dentro del bulbo. La microsporogénesis sigue un patrón de tipo sucesivo, común en las monocotiledóneas. El proceso es asincrónico entre yemas de una misma inflorescencia (indeterminado o acrópeto), anteras de una misma flor, y entre el ápi (more) ce y la base de la misma antera (basípeto). Las anteras poseen estomas, el tapete es de tipo ameboide o periplasmodial y las células del endotecio no desarrollan engrosamientos en la pared, al contrario de lo que ocurre en la mayoría de los representantes de la familia. El tapete invade tempranamente la cavidad de la antera y produce pollenkit, una secreción amarillenta de tipo lipídico que se deposita sobre los granos de polen maduros confiriéndoles propiedades adherentes. Las micrósporas son uninucleadas, con patrón de ornamentación incipiente al momento de ser liberadas de la tétrada. Los granos de polen son monocolpados, como en la mayor parte de las monocotiledóneas; la exina tiene un patrón de ornamentación denso en los extremos y un retículo más laxo en la zona central. Al momento de la dehiscencia de la antera el polen es binucleado, condición que se considera ancestral en las angiospermas junto a la presencia de estomas en las anteras. Abstract in english An embryological investigation was undertaken for the first time in a member of the genus Hymenocallis, to describe the microsporogenesis in Hymenocallis caribaea. Meiosis takes place when flower buds are still inside the bulb. Microsporogenesis follows a successive type, common in monocots. The process is asynchronous between buds of the same inflorescence (undeterminate or acropetal), anthers of the same flower, and between the apex and the base of each anther (basipeta (more) l). Anthers have stomata, the tapetum is ameboid, or periplasmodial, and the endothecium doesn?t develop wall thickenings, contrary to what happens in most family representatives. Tapetum invades anther cavity early and produces pollenkit, a yellowish lipid secretion which is deposited over the mature pollen grains giving them adherent properties. When released from tetrads, microspores are uninucleate and show an undeveloped pattern of wall sculpturing. Pollen grains are monosulcate as in most monocots; the exine shows a dense sculpturing pattern at the ends and a loose reticulum at the central zone. At the time of anther dehiscence, pollen is binucleate, a condition which is considered ancestral within the angiosperms as is the presence of stomata in the anthers.

Abstract in portuguese Este experimento, realizado na região de Jaboticabal (SP), utilizou uma cultura de pêssego (Prunus persica L.), durante a sua florada com a finalidade de verificar a atuação dos insetos visitantes nas flores na produção de frutos. A concentração média de açucares no néctar e a quantidade média produzida por dia de néctar é de 27,9% e de 3,2 mg, respectivamente. O peso médio das anteras por flor foi de 1,59 mg. A abelha Apis mellifera (73%) foi o principal i (more) nseto visitante seguida da Trigona spinipes (17%) e Xylocopa sp (4%). Observou-se a presença de beija-flores (6%), coletando néctar. A freqüência máxima das abelhas A. mellifera, para coleta de néctar e pólen, ocorreu as 12 horas. O número de frutos resultantes do tratamento em que as flores recebiam as visitas foi 14% maior que no tratamento em as flores não eram visitadas. Do total de frutos colhidos no tratamento coberto (sem visitas), 82% apresentaram-se perfeitos, com boa formação e simetria. No tratamento descoberto, 90,2% apresentaram-se com boa formação, havendo diferença estatística entre os dois tratamentos. Abstract in english This experiment, accomplished in the area of Jaboticabal (SP), it used a peach culture (Prunus persica L.), during yours bloomed with the purpose of verifying the insects visitors performance in the flowers in the production of fruits. The medium concentration of sugar in the nectar and the medium amount produced a day of nectar it is of 27,9% and of 3,2 mg, respectively. The medium weight of the anthers for flower was of 1,59 mg. The honeybee Apis mellifera (73%) it was (more) the principal insect visitor followed by the Trigona spinipes (17%) and Xylocopa sp (4%). The presence of hummingbird was observed (6%), collecting nectar. The maximum frequency of the bees A. mellifera, for nectar collection and pollen, happened the 12 hours. The number of resulting fruits of the treatment in that the flowers received the visits it was 14% larger than in the treatment in the flowers were not visited. Of the total of fruits picked in the covered treatment (without visits), 82% came perfect, with good formation and symmetry. In the discovered treatment, 90,2% came with good formation, having difference statistics among the two treatments.

The unfolded protein response (UPR) is a regulatory system to maintain the homeostasis of ER functions. Here we report a comparison of express levels of UPR relevant genes in Aspergillus oryzae between solid-state and submerged cultivation. The results were that up-regulation of the UPR mechanism in solid-state culture was higher than in submerged culture (heat-shock or non-stress conditions). This might have been a result of changing culture conditions.   

The intensity of lymphocyte proliferation in response to pokeweed mitogen depends on cortisol level in the peripheral blood in the early post-traumatic period of penetrating eye injury. Lymphocyte proliferation in 72- and 96-h cultures from patients with high levels of endo genous hormone was suppressed. In 120-h cultures, the intensity of proliferation remains unchanged. Lymphocyte blast transformation was increased in 120-h cultures from patients with normal cortisol concentration and remained unchanged in case of low cortisol level.

To improve rat embryo culture conditions, responses of Wistar 2-cell embryos from 2 breeders to oxygen tension (5 vs 20%) and bovine serum albumin (BSA) (0 vs 3 mg/ml) were examined using rat 1-cell embryo culture medium (mR1ECM). Supplementation of 3 mg/ml BSA significantly stimulated and accelerated development to the blastocyst and expanded blastocyst stages during 72 and 96 h culture, while reduced oxygen tension stimulated cell division. Fetus development after transfer of blastocysts obtained from 72 h culture under 5% O2 with BSA was significantly higher than those cultured under atmospheric oxygen without BSA. However, the nuclear numbers of in vitro cultured blastocysts and fetus development after embryo transfer were still significantly lower than in vivo developed blastocysts, indicating the current culture condition is still suboptimal.   

The plasmid profiles of 619 cultures of Bacillus anthracis which had been isolated and stored between 1954 and 1989 were analyzed using the Laboratory Response Network real-time PCR assay targeting a chromosomal marker and both virulence plasmids (pXO1 and pXO2). The cultures were stored at ambient ...

Many xenobiotic compounds are biodegradable in laboratory bacterial cultures, but results in the environment are not as reassuring. Actual biodegradation rates of aromatics under natural conditions may be very low. This study mimicked limiting conditions of oxygen phosphate and nitrogen in chemostat cultures of Pseudomonas putida and studied the inducibility of TOL plasmid pathway in response to the nonmetabolizable inducer-o-xylene.

Two opposite effects of actinomycin D on antibody synthesis have been studied in organ cultures of rabbit lymph node fragments. These cultures were prepared from previously primed rabbits and stimulated with antigen(s) on day 0 to yield a secondary response, whose inductive phase extended to about ...

The growth response of normal and human papillomavirus (HPV)-transformed cervical keratinocytes to interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha was investigated in monolayer and organotypic raft cultures. The proliferation rates of monolayer cultures were assessed by [3H]TdR incorpor...

Recent linguistic research has shown that cultural processes operating over an extended timescale may be responsible for many aspects of syntax. Other evidence from artificial language learning studies indicates a strong bias for systematicity, potentially conflicting with these cultural accounts o...

Coastal barrier systems consist of two primary components, or behavioral elements. The most obvious component is the seaward system, or active shoreface, whose evolution is dominated by transport and reworking by waves. The shoreface is affected by both extreme (storm) events and the longer-term effects of less extreme wave conditions. The terrestrial system, the barrier landward of the shoreline, can be controlled by a number of different processes, primarily storm overwash, aeolian dune development, and, within the Anthropocene, human interventions. The terrestrial and shoreface systems are connected at the coast, they both tend to change dramatically during storms, and, similarly, are slowly active between extreme events. However, other than the coupling at the shared boundary of the shoreline, the dynamics and processes of these two systems are largely independent of one anther. Dynamic coupling of these two systems within a morphodynamic feedback framework allows a better understanding of how the coupled system behaves over long timescales (decades, centuries, to millennia). An extremely simplified ‘panel’ model demonstrates that a hypothesized equilibrium rollover model of barrier response to sea-level rise (as assumed by all geometric models) would require that these disparate systems evolve to a state where they become directly coupled, with the overwash, or terrestrial, component driving the long-term evolution of the shoreface, even determining the direction of long-term sediment fluxes. This simple modeling framework also illuminates the characteristic temporal scales and sediment transport rates for long-term equilibrium behaviors (one where sea level rise does not drown barrier islands), demonstrating the potential for instabilities within the coupled system, either in response to changes in rates of sea level rise or changes to the backbarrier slope. A more detailed modeling framework allows investigation of different relationships for barrier overwash, including modeling of the impacts of aeolian dune development and human interventions on the subaerial barrier. These modeling investigations, by demonstrating the importance of overwash on long-term, large-scale coastal evolution, motivate the need to quantify the timing and flux of overwash in natural environments. Field investigations using ground-penetrating radar (GPR) can provide a means to quantify sediment volumes of previous overwash events. We present, along with the theoretical results, preliminary results of GPR surveys of overwash fans along the active barriers of Cape Cod and Martha’s Vineyard, Massachusetts, demonstrating the capability to use gridded high-density geophysical surveys to constrain overwash fluxes within a local framework. These field investigations provide a means to test the general predictions of the morphodynamic models, shedding further light on long-term barrier evolution.

summary, changes in immunological parameters were shown to be occurring in cells obtained from in-flight dissections of ... alterations in immune responses induced by space flight. ..... cultures of C3H/HeJ murine thymus (2 month old males).

The macrophage migration inhibition test has been used to study the immune responses of guinea-pigs immunized with injections of whole cells of both an allogeneic and a syngeneic hepatoma grown as established cell lines in tissue culture.

Hepcidin is an iron regulatory hormone expressed in the retina. In the present study, evidence from mice and tissue culture suggest that hepcidin is upregulated in response to increased retinal iron levels and normally serves to prevent retinal iron excess.

Physiological concentrations of silicon (Si) have been shown to affect cellular immune function in cultured lymphocytes. Thus, experiments were performed with rats to determine whether (Si) deprivation influences bone turnover and the inflammatory response by altering circulating cytokines and blood...

The smut genus Thecaphora contains plant parasitic microfungi that typically infect very specific plant organs. In this study, we describe a new species of Thecaphora from Oxalis lanata var. rosea (Oxalidaceae) in the Cape Floristic Region of South Africa. Molecular phylogenetic reconstructions based on large subunit ribosomal DNA sequence data confirmed the generic placement of the fungus and confirmed that it represents an undescribed species for which the name T. capensis sp. nov. is provided. The closest known sister species of the new taxon is T. oxalidis that infects the fruits of Oxalis spp. in Europe, Asia and the Americas. In contrast, T. capensis produces teliospores within the anthers of its host. This is the first documented case of an anther-smut from an African species of Oxalis and the first Thecaphora species described from Africa. PMID:20396584

Micro- and megasporogenesis as well as male and female gametogenesis in genus Agapanthus are reported for the first time. The results are as follows: The anther wall, whose development is of dicotyledonous type, consists of four layers: the epidermis, the endothecium, one or two middle layers, and the secretory tapetum. Fibrous thickenings are developed in the endothecium when shed. Successive cytokinesis during microsporogenesis results in a decussate tetrad of microspores. Ellipsoidal pollen grains are 2-celled when anthers dehisce, with monosulcate extending to polar area; pollen ornamentation is pitted and reticulate. Ovary is superior and trilocular, with axial placenta. The ovule is anatropous, bitegmic, and tenuinucellate. The archesporial cell below nucellus epidermis functions dir...

Summary. Western blot analysis indicated the presence of two epitopes recognized by the anti-arabinogalactan protein antibodies JIM13 and LM2 and the absence of the JIM4 epitope in mature tobacco anthers. Immunoenzyme localization of arabinogalactan proteins (AGPs) with JIM13 showed that AGPs accumulate mainly at the early stages of anther development. AGP content and distribution were also investigated at the ultrastructural level in pollen tubes grown in vivo and in vitro. Abundant AGPs were present in the transmitting tissue of styles, and the AGP content of the extracellular matrix changed during pollen tube growth. In pollen tubes, immunogold particles were mainly distributed in the cell wall and cytoplasm, especially around the peripheral region of the generative-cell wall. ?-D-Gluco...

Gametophyte development is an important process in flowering plants. Seeds are the primary organs for harvesting, and therefore spikelet fertility is especially vital for rice yield. Here, we report the characterization and genetic mapping of a novel rice male and female sterile mutant, mfs1, which was generated from a transgenic line of japonica rice variety Zhonghua 9. Anther transverse sections indicate that the microspores in these plants were abnormally degenerated at the meiotic leptotene stage, while the tapetum layer was not degenerated and remained present; therefore, no functional pollen grains formed in the mutant anther locules. In addition, the majority of mutant embryo sacs did not undergo differentiation or the differentiation was incomplete, resulting in the absence of func...

Effects of epidermal growth factor (EGF) on the expression of alpha(2)-adrenergic responses were examined in primary cultures of adult rat hepatocytes. The alpha(2)-responses were assessed by the inhibition of the rate of forskolin-stimulated cAMP formation by the selective alpha(2)-adrenergic agonists, oxymetazoline and UK-14304. Hepatocytes cultured with EGF (20 ng/ml) at a high cell density (1.0 x 10(5)/cm2) showed almost no response to the alpha(2)-adrenergic agonists, oxymetazoline and UK-14304 (1-100 mu M). In contrast, when cultured at a low cell density (3.3 x 10(4) cells/cm2) with EGF, forskolin-stimulated cAMP production was inhibited by oxymetazoline and UK-14304 in a dose-dependent manner. The alpha(2)-response was blocked by the alpha2-antagonist yohimbine (10 mu M). It was also reversed by treatment of hepatocytes with pertussis toxin (100 ng/ml). In addition, the effects of EGF on the appearance of alpha(2)-responses were almost completely inhibited by treatment of the hepatocytes with genistein (10 mu M) or cytochalasin B (10 mu M). The alpha(2)-response was abolished when cycloheximide (5 mu M) was added to the cultures. These results demonstrate that when cultured at a low cell density with EGF, adult rat hepatocytes acquire a significant alpha(2)-adrenergic response. The expression of this alpha(2)-response is associated with de novo protein synthesis. PMID:8860950

When the female plant of Silene latifolia is infected with the smut fungus Microbotryum violaceum, its rudimentary stamens develop into anthers which contain fungus teliospores instead of pollen. To identify genes required for maturation of anthers in S. latifolia, we performed a cDNA subtraction approach with healthy male buds and female buds infected with M. violaceum. We isolated five cDNA clones, which were preferentially expressed in healthy male buds during stages associated with a burst in tapetal activity. These five cDNAs are predicted to encode a mandelonitrile lyase protein (SlMDL1), a strictosidine synthase protein (SlSs), a glycosyl hydrolase 17 protein (SlGh17), a proline-rich protein APG precursor (SlAPG), and a chalcone-synthase-like protein (SlChs). All five genes showed expression in both healthy and fungus-infected male buds, but not expressed in either healthy or infected female buds. The first three genes were highly expressed in both tapetum and pollen grains while the last two genes were expressed only inside the tapetum of male flower buds. Phylogenetic analysis results showed that SlChs and SlGh17 belong to anther-specific subgroups of chalcone-synthase-like genes and glycosyl hydrolase 17 family genes, respectively. Our results suggest that the isolated five genes are related to the fertility of the anther leading to the development of fertile pollen. The smut fungus was not able to induce the expression of the five genes in the infected female buds. This raises the possibility that these genes are under the control of master gene(s) on the Y chromosome.   

Summary A new species of Chlorophytum is described and illustrated. The species is allied to Chlorophytum arundinaceum Baker but differs in its roots possessing root buds and ellipsoid tubers, its pedicels being jointed above the middle, perianth lobes 3 ? 5-nerved and filaments shorter than anthers. In addition to sexual reproduction, the species propagates through formation of buds on roots, which grow into new plants. The chromosome number of the species is 2n = 42.

Aegilops tauschii Coss. (syn Ae. squarrosa L.) is a wild diploid wheat species. It has a wide natural species range in central Eurasia, spreading from northern Syria and Turkey to western China. Ae. tauschii is known as the D genome progenitor of hexaploid bread wheat. The genealogical and geographical structure of variation of morphological traits was analyzed using a diverse array of 205 sample accessions that represented the entire species range. In total, 27 traits, including anther and pistil shape and internode length, were examined in this study. Large-scale natural variation was found for all examined traits. Geographically, significant longitudinal clines were detected for anther size, internode length and spike size and shape. Anthers tended to be small in accessions from the eastern region. Internodes also tended to be short, whereas spikes tended to be long in accessions from the eastern region. Spikelet density per spike tended to be high in the eastern habitats. In the process of west-to-east dispersal, Ae. tauschii underwent extensive morphological, genetic and ecological diversification that produced the variation seen among today’s natural populations.   

Several genes encoding transcription factors have been shown to be essential for male fertility in plants, suggesting that transcriptional regulation is a major mechanism controlling anther development in Arabidopsis. DYSFUNCTIONAL TAPETUM?1 (DYT1), a putative bHLH transcription factor, plays a critical role in regulating tapetum function and pollen development. Here, we compare the transcriptomes of young anthers of wild-type and the dyt1 mutant, demonstrating that DYT1 is upstream of at least 22 genes encoding transcription factors and regulates the expression of a large number of genes, including genes involved in specific metabolic pathways. We also show that DYT1 can bind to DNA in a sequence-specific manner in vitro, and induction of DYT1 activity in vivo activated the expression of the downstream transcription factor genes MYB35 and MS1. We generated DYT1-SRDX transgenic plants whose fertility was dramatically reduced, implying that DYT1 probably acts as a transcriptional activator. Furthermore, we used yeast two-hybrid assays to show that DYT1 forms homodimers and heterodimers with other bHLH transcription factors. Our results demonstrate the important role of DYT1 in regulating anther transcriptome and function, and supporting normal pollen development. PMID:22775442

The Arabidopsis AtSUC1 protein has previously been characterized as a plasma membrane H+-sucrose symporter. This paper describes the sites of AtSUC1 gene expression and AtSUC1 protein localization and assigns specific functions to this sucrose transporter in anther development and pollen tube growth. RNase protection assays revealed AtSUC1 expression exclusively in floral tissue, which was confirmed by analyses of AtSUC1 promoter-beta-glucuronidase (GUS) plants. In situ hybridizations identified AtSUC1 expression in anther connective tissue, in funiculi and in fully developed pollen grains. Indirect immuno-fluorescence analyses with anti-AtSUC1 antiserum confirmed AtSUC1 protein localization in the connective tissue and funiculi. In mature pollen grains, however, despite high AtSUC1 mRNA levels no AtSUC1 protein was found. Only after pollination of stylar papillae was AtSUC1 protein detected inside the pollen and later inside the growing pollen tubes, suggesting a translation of pre-existing AtSUC1 mRNA after pollination. Pollen germination analyses underlined the important role of sucrose for pollen tube growth. The data presented suggest a role of AtSUC1 in the controlled dehiscence of Arabidopsis anthers. It is postulated that an important function of AtSUC1 is the cell-specific modulation of water potentials. PMID:10476074

Recent work on biculturalism has made theoretical and methodological inroads into our understanding of the relation of cultural processes with psychological functioning. Through the use of cultural priming methodologies, investigators have demonstrated that biculturals, or individuals who have experienced and identify with more than one culture, can switch between various “cultural frames of reference” in response to corresponding social cues (Hong, Morris, Chiu, & Benet-Martinez, 2000). Drawing on this work on the cognitive implications of biculturalism, the purpose of the present study was to examine the assumption that independent and interdependent self-construals are associated with the motivational standards of self-enhancement and self-criticism, respectively. Mor...

Teacher educators have a civic responsibility to prepare novice teachers to foster relationships across cultural, racial, and socioeconomic divides. Care ethics acknowledges this imperative and context's role in determining varied meanings of care. The voices of teachers of color can help us to understand the demands of care across differences. This study presents findings from interviews with teachers of color whose preparation included an introduction to care ethics. Five themes emerged: the importance of assuming students' caring intentions, the importance of teachers' persistence in relationship with students, connection to students' cultures, connection of students' cultures to an academic culture, and acknowledgment and confrontation of racism. (Contains 1 note.)

Culturally responsive science teaching is using knowledge about the culture and life experiences of students to structure learning that is conducive to their needs. Understanding what teachers need to prepare them to be culturally responsive is a matter of continuous debate. As the focus of multicultural education ventures farther away from its roots, advocating the civil rights of historically oppressed groups, concerns about the gravity of racial inequity on schooling continues. How will this shift in focus influence teachers? capacity to accommodate students? needs resulting from racial inequities in this society, particularly African American students? What knowledge is essential to their effectiveness? This qualitative study examined the instructional practices of two effective middle...

The mitogen phytohemagglutinin (PHA) works well in both human and cynomolgus monkey (Macaca fascicularis) lymphocyte cultures to stimulate T cell proliferation. T cells from rhesus monkeys (Macaca mulatta) are less responsive than human cells, producing few metaphases when thousands are required, e.g. in biological dosimetry studies. We show that staphylococcal enterotoxin A (SEA), one of the most potent mitogens known, at a concentration of 0.5 microgram/ml stimulated peripheral lymphocytes to grow with a mitotic index (MI) averaging 0.13 metaphases/cell in old, irradiated rhesus macaques. This was significantly greater (p < 0.001) than that produced by PHA (MI < 0.01) in lymphocytes from the same animals. Whole blood was cultured for 96, 120 and 144 h for five irradiated individuals and for two controls. All cells cultured with SEA produced a high MI with a peak response at 120 h whereas the same cultures showed low MI for each PHA stimulated culture.

Rosiglitazone (RSG) has been known to play a role in the modulation of inflammatory responses. Therefore, we sought to elucidate the underlying molecular mechanism by which RSG regulates the development of rheumatoid arthritis. Firstly, we examined the preventive effect of RSG on the inflammatory mediators induced by spheroid culture of synovial sarcoma SW982. Expression of proinflammatory cytokines under spheroid culture was more elevated than that under monolayer culture while RSG abolished inflammatory responses. The upregulation of inflammation-related genes by spheroid culture was closely associated with NFkappaB (NFkB) activation. Also, activation of p38 and c-Jun N-terminal kinase (JNK) by spheroid culture was abrogated with RSG treatment. Lastly, it was demonstrated that RSG reduce...

Abstract in spanish Los objetivos del trabajo fueron determinar la época de maduración del polen y del estigma y establecer el tipo de polinización de la uchuva en condiciones de invernadero con cinco genotipos de uchuva. La investigación se realizó en la Granja Botana de la Universidad de Nariño (2.820 msnm, 13°C, precipitación de 800 mm/año y humedad relativa de 82%). P. peruviana tomó 37 días para la apertura floral, la cual se efectuó entre las 7:00 a.m. y las 10:00 a.m. En e (more) l 85% de las flores la primera antera fue dehiscente al día siguiente de la apertura floral. Los granos de polen tuvieron 97% de viabilidad a los 35 días. El polen maduró antes de la antesis y el estigma fue receptivo antes de la apertura de la flor. Tanto el polen como el pistilo maduraron a los 35 días de desarrollo, dos días antes de la apertura floral; la receptividad del pistilo se presentó dos días antes de la antesis, fenómeno que restringe la autopolinización. En invernadero, la ausencia de vectores influyó en la polinización de P. peruviana. En flores emasculadas y sometidas a libre polinización se presentó baja formación de frutos y semillas, existió respuesta diferencial a la polinización dentro del invernadero entre las muestras evaluadas. P. peruviana presentó polinización mixta con 54% de polinización cruzada. Abstract in english The objectives of the study were to determine the time of pollen and stigma ripening and to establish the kind of pollination in Physalis. This research was conducted in the Botana Farm at the University of Nariño, Colombia ( 2820 m a s l, 13 ° C, pluvial precipitation of 800 mm / year and relative humidity of 82%). P. peruviana took 37 days to anthesis, which took place between 7:00 and 10:00 am. In 85% of the flowers the first anther dehiscent was the day after the op (more) ening of the flower. The pollen grains showed 97% of viability to 35 days. Pollen matured before anthesis and stigma was receptive before the opening of the flower. Both, pollen and the pistil matured to 35 days of development, two days before anthesis; receptivity of the pistil was filed two days prior to anthesis, a phenomenon that restricts the self. The absence of vectors influenced pollination of P. peruviana. In emasculate flowers and subjected to self-pollination is introduced low fruit and seed formation, there was a differential response in pollination among samples inside the greenhouse. P. peruviana showed mixed pollination with 54% of cross-pollination.

Barley microspores from five field-grown breeding lines were isolated using an ultra-speed blender and the effect of co-culture with young florets was investigated. Floret co-culture in the induction stage increased the formation of MCS, ELS and green plant regeneration. The florets of teraploid plant were more effective than ones of diploid plant. For line S23, co-culture with florets from tetraploid plants gave rise to 2.6 and 7.8 times more MCS and ELS, respectively, than non-co-culture control, whereas co-culture with florets from diploid plants resulted in 1.8 and 6.1 times more MCS and ELS, respectively, than non-co-culture control (Table?2). Florets subjected to cold treatment for 10?20 days induced a greater response than fresh ones, and florets with uninucleate microspores surpass...

Calls for cultural sensitivity in the design and implementation of human services programs have become a standard response to the increasing diversity among the families and communities being served. In this article, we take a critical look at the construct, using data from a multi-year evaluation of a statewide family support program. We examine how selected, locally implemented programs operationalize the state programs cultural sensitivity standard, using both etic and emic approaches; that is, we apply extant cultural competence definitions to assess program cultural sensitivity and document how the program staff articulated their approach to the cultural sensitivity. Findings suggest that programs focus more directly on the immediate cultural relevance of their services, rather than o...

The purpose of this study is to examine the perceptions of elementary school teachers about the sensitiveness of principals, teachers, and curriculum on multicultural education. Education provides the transmission and the advancement of its culture while it is developing and enhancing the common values, the integrity and the progress of multicultural society (Sahin, 2006). If the society has multi-ethnic culture, the educational policy should cover all kinds of multi-ethnic cultures to exchange cultural values each other. The findings of this study indicate that Turkish educational system ignores multiculturalism in their schools. Curriculum does not cover cultural differences. Principals and teachers performing their responsibility relatively show respect different cultures even if it is not at the expected level. (Contains 5 tables.)

This study on the impact of educational interventions on organizational culture is an evaluation of a major educational initiative undertaken by an urban federal agency, namely the National Aeronautics and Space Administration's Langley Research Center (NASA-LaRC). The design of this educational evaluation captures the essence of NASA-LaRC's efforts to continue its distinguished and international stature in the aeronautical research community following the Challenger tragedy. More specifically, this study is an evaluation of the educational initiative designed to ameliorate organizational culture via educational interventions, with emphasis on communications, rewards and recognition, and career development. After completing a review of the related literature, chronicling the educational initiative, interviewing senior managers and employees, and critically examining thousands of free responses on employee perceptions of organizational culture, it is found that previous definitions of organizational culture are more accurately classified as manifestations of organizational culture. This research has endeared to redefine 'organizational culture' by offering a more accurate and diagnostic perspective.

Apoptosis plays an important role in eliminating cells from populations when cells have been exposed to UV irradiation and damaged. Studies of cells in culture have provided some details of the mechanisms involved when stress response genes act after exposure to UV irradiation and other environmental stresses. However, little is known about the responses of intact sections of human skin growing in organ culture to UV irradiation. In the work reported here, it was found that the response of organ-cultured human skin after exposure to UV irradiation is different than the response of cultured cells. At wavelengths below 300 nm, the action spectrum obtained from organ-cultured skin samples showed a lower sensitivity than that observed at 300 nm, indicating that the overlying stratum corneum and upper epidermal cell layers had probably caused a selective absorption of incident UV radiation at some wavelengths. At 3 hours after UV irradiation, p53 was phosphorylated at Ser 15 and Ser 46, and accumulated in the cell nuclei, notably after exposure to 280-320 nm wavelengths. Accumulations of Bax, active Caspase-3 and cleaved PARP were detected in apoptotic cells at 24 hours post-exposure, along with a reduction of Bcl-2 levels, notably after exposure to 300-365 nm light. This difference in apoptotic responses may result from the characteristics of the different irradiation wavelengths used, and from details in the skin's structure. The data obtained in this study using an organ-culture system utilized direct measurements of the biological effects of different wavelengths of UV lights.   

Australia is a culturally and linguistically diverse country with a population derived from over 140 countries and including 240 language groups. Reflecting this, there has been a significant increase in cultural diversity among undergraduate dental students. It has been recognized that in order for dental students to interact and respond effectively to the diverse cultural needs of their patients, students themselves must be aware of cultural differences and respect patients' worldviews. In response to this challenge, dental students will need to have the theoretical knowledge to understand culturally-influenced health behaviours as well as the ability to communicate effectively with culturally diverse patients. Currently, the culture of dental students contrasts with the patients they treat, which may in turn affect the interaction between dental students and their patients. Given this context, new graduates need both to effectively communicate with patients from diverse communities and have an understanding of culturally influenced health behaviours. It has been proposed that dental graduates need to improve their knowledge of a variety of cultural values, beliefs, practices and attitudes. The literature in the area of cultural awareness and education for oral health professionals concentrates on both exploring health professionals' knowledge and attitudes toward transcultural care or the need for transcultural training. This paper provides an overview of the transcultural issues in oral health care which might confront dental students when treating culturally diverse patients. It will also discuss possible modifications to the dental curriculum to ensure that the future oral health workforce understands the complex health care needs of a multicultural society. This information will give planners and stakeholders an insight into the nature of the cultural issues which future dentists are likely to encounter while treating patients from diverse cultural backgrounds. This would help to establish the need to incorporate transcultural awareness modules to enhance quality of care and to respond effectively and sensitively to cultural issues. PMID:22998318

Persson's (2012a) target article calls for a cultural sensitive research paradigm in the science of giftedness. It charts the potential threats to research validity affected by cultural bias having implications on study and practice in gifted education. The eight recommendations heading under: (1) mindset and habits; (2) research skills; and (3) self-knowledge and cultural competence proposed by Persson (2012a) are very important. The first seven, considered by researchers, will definitely improve credibility and contribute to developing culture-sensitive research paradigms. The eighth recommendation involving the acquisition of cultural competence learning about own and other cultural beliefs, and value patterns is the most difficult to accomplish. In this commentary, the author poses some questions concerning: (1) cultural clusters constituting countries; (2) fundamental perceptions differences; and (3) cultural relativism in research and teacher education. In addition, the author attempts to answer the following question: What do cultural relativism and culturally responsible research have to do with teacher certification programmes and gifted education services?

The objective of this study was to examine the in vitro responsiveness of cultured luteinized human granulosa cells over time to insulin-like growth factor 1 (IGF-1), human follicle-stimulating hormone (FSH), and human chorionic gonadotropin (hCG) for the induction of aromatase activity. Granulosa cells were retrieved from preovulatory follicles in patients undergoing in vitro fertilization. Cells were cultured for a period of 72 hours or 10 days. The ability of hCG, human FSH, and/or IGF-I to induce aromatase activity was assayed by the stereospecific release of tritium from (1B-3H)androstenedione. Short-term cultures (72 hours) demonstrated a marked rise in aromatase activity in response to human FSH and IGF-I, whereas a smaller response to hCG was observed. In contrast, 10-day cultures demonstrated responsiveness predominantly to hCG rather than human FSH for the induction of aromatase activity with no remarkable effect of IGF-I. Luteinized human granulosa cells undergo a transformation from an initial human FSH and IGF-I responsive state to an hCG responsive state in long-term cultures.

The primary and secondary cytotoxic T cell responses of C57Bl/6 (H-2b) mouse spleen cells to P-815 membrane fragments (H-2d) were examined. The primary response required the addition of a supernatant from mitogen-stimulated spleen cells, or costimulator, to the culture medium. Costimulator had littl...

In plant/parasitic plant interaction, little is known about the host plant response before the establishment of the parasite within the host. In the present work, we focused on host responses to parasitic plant, O. ramosa in the early stage of infection. We used a co-culture system of A. thaliana su...

Abstract in portuguese No manejo do cultivo da pinha (Annona squamosa), a polinização artificial é uma prática preconizada para, obter maior pegamento dos frutos bem como uniformização do formato dos mesmos. Nesse sentido, conduziu-se este trabalho, com o objetivo de avaliar a viabilidade dos grãos de pólen de flores de pinheira em diferentes horários de coleta. O pólen foi obtido a partir de flores no estádio funcionalmente estaminada. Foram avaliados oito horários de coleta de pó (more) len: zero hora, 1 hora, 2 horas, 3 horas, 4 horas, 5 horas, 6 horas e 7 horas da manhã. Foi utilizado meio de cultura padrão para germinação de pólen, com concentração de 10% de sacarose. As flores foram coletadas nos horários estabelecidos e os grãos de pólen foram retirados das anteras com auxílio de um pincel número 2 e em seguida inoculados em placas de Petri contendo o meio de cultura. O delineamento experimental foi inteiramente casualizado com quatro repetições, sendo cada parcela experimental constituída por duas placas Petri. Foram contados 100 grãos de pólen por placa. Após 6 horas de inoculação, os grãos de pólen foram visualizados sob lupa. Foram considerados germinados os grãos de pólen que possuíam tubo polínico com tamanho igual ou superior ao diâmetro do próprio pólen. Não foram observadas diferenças significativas entre o horário de coleta dos grãos de pólen. A percentagem média da germinação variou de 46,75% a 53,62% dos grãos de pólen germinados. Abstract in english In the management of the sugar apple (Annona squamosa) crop, the artificial pollination is a preconized practice to obtain higher establishment of the fruits as well as their standardization. This study was carried out to evaluate the viability of sugar apple pollen grains at different collecting times. The pollens were obtained from the flowers at the functional staminate stage. The pollen grains were collected every hour, starting from 00:00am and ending at 07:00am, tot (more) alizing 8 collections. The standard culture medium with 10% sucrose was used for the germination of the pollen. The flowers were collected at the scheduled hours and the pollen grains were taken from the anthers, by using a brush # 2. They were then inoculated on Petri dishes containing the culture medium. The completely randomized experimental design was used with four replicates, and each plot was constituted by two Petri dishes. One hundred pollen grains were counted in each Petri dish. After six hours of inoculation, the pollen grains were visualized under magnifying glass. Those pollen grains containing the pollinic tube with the same or higher size than their own pollen diameter were considered germinated. No significant differences were found among the different hours of the pollen grain collections, and the germination percentage ranged from 46.75% to 53.62%.

This study examines differences in tumour cellular response using clonogenic cell survival between uniform and non-uniform irradiation. Cells were irradiated with a 6 MV x-ray intensity-modulated beam, in a single large flask (i.e. intercellular communication is possible) or in three small flasks (i.e. intercellular communication is inhibited across the dose gradient). For non-small-cell lung cancer and melanoma cell lines, the dose response over the entire cell culture was significantly different between freely communicating cell cultures and those with inhibited communication across the dose non-uniformity. Communicating cells exhibited poorer survival in the low dose region of the field but improved survival in the high dose region. In general, the response to non-uniform irradiation appeared to 'average out' over the entire cell culture. This was not seen when intercellular communication was inhibited. The results add strength to the body of evidence regarding bystander effects and the inter-dependence of cellular response.

Endothelial cells (ECs) that line the inner surface of blood vessels are continuously exposed to shear stress induced by blood flow, in vivo, and shear stress affects the intracellular calcium ([Ca2+]i), which initiates cellular responses. Although shear-stress induced [Ca2+]i responses shortly (within 10min) after initiation of flow have been studied, the effects of long-term exposure (24hr) of shear stress on [Ca2+]i responses have not. Here, we studied the effect of long-term exposure of shear stress on [Ca2+]i responses in cultured ECs by using a confocal laser microscope and calcium indicator (Calcium Green-1/AM). At the initiation of shear stress of 2Pa (0hr), 27% of the cells exhibited [Ca2+]i responses. This percentage gradually decreased with increasing exposure time, reaching about 4% after 24hr of exposure. These data indicate that long-term shear-stress exposure affects [Ca2+]i responses in cultured ECs.   

The purpose of our study was to examine if lipopolysaccharide (LPS) from Porphyromonas gingivalis (P.g.) modifies the vasomotor responses to Endothelin-1 (ET-1) and Sarafotoxin 6c (S6c) in rat coronary arteries. The arteries were studied directly or following organ culture for 24 h in absence and presence of 2.5EU/ml LPS. The contractile responses of coronary arteries were investigated by using the selective ETB receptor agonist S6c (1 pM-0.3 µM) and ET-1 (1 pM-0.3 µM). The functional studies demonstrated an augmented contractile response only to S6c in isolated rat coronary arteries after organ culture (with or without LPS). These contractile responses by S6c were blocked by the selective ETB receptor antagonist BQ788 in both vessel groups. The augmented contractile response to S6c was supported by immunohistochemistry, where a significant increase in fluorescence intensity for ETB receptors in smooth muscle cells was observed after organ culture. The presence of LPS in the culture medium significantly increased the sensitivity of endothelium-intact coronary artery to S6c as compared to endothelium-denuded segments. Our results showed a significant increase in both ETB receptor protein levels and S6c-induced maximal contraction in coronary arteries upon 24 h of organ culture, which was further sensitized by LPS.

Dilution assays are quantal dose-response assays that detect a positive or negative response in each individual culture within groups of replicate cultures that vary in the dose of cells/organisms tested. We propose three jackknife versions of the maximum likelihood estimator of the unknown parameter, i.e., the frequency of a well-defined cell within the context of limiting dilution assays or the density of organisms within the context of serial dilution assays. The methods have been evaluated with artificial data from extensive Monte Carlo experiments. As a result of these experiments and theoretical considerations, the jackknife version based on deleting one individual culture at a time is proposed as the statistical procedure of choice. The next best method is the jackknife version based on leaving out the same replicate from each of the culture groups at a time. PMID:3233248

Abstract Osteoblasts (OBs) exert a prominent regulatory effect on hematopoietic stem cells (HSCs). We evaluated the difference in hematopoietic expansion and function in response to co-culture with OBs at various stages of development. Murine calvarial OBs were seeded directly (fresh) or cultured for 1, 2, or 3 weeks prior to seeding with 1000 Lin-Sca1-+-cKit+ (LSK) cells for 1 week. Significant increases in the following hematopoietic parameters were detected when comparing co-cultures of fresh OBs to co-cultures containing OBs cultured for 1, 2, or 3 weeks: total hematopoietic cell number (up to a 3.4-fold increase), total colony forming unit (CFU) number in LSK progeny (up to an 18.1-fold increase), and percentage of Lin-Sca1+ cells (up to a 31.8-fold increase). Importantly, these studi...

The goal of this work was to investigate the influence of Baltic cyanobacteria Anabaena variabilis and Nodularia spumigena cells and cell-free filtrates on the growth of green algae Chlorella vulgaris.We have demonstrated that Anabaena variabilis and Nodularia spumigena caused allelopathic effects against microalgae.The cyanobacterial and microalgal cultures were provided on liquid medium, in 22 °C at continuous light. Cell-free filtrates were obtained by centrifugation and filtering aliquots of cyanobacterial cultures (including cultures in exponential and stationary phase of growth).Growth response of free cells (batch culture technique) and immobilized cultures (in alginate beads) of the unicellular green algae to cyanobacteria allelochemicals were tested and compared. In this experiment Anabaena variabilis supressed the growth of microalgae compared to control samples. Nodularia spumigena stimulated the growth of Chlorella vulgaris in most cases, however both positive and negative effects were observed.

Mixed lymphocyte culture (MLC) responses and lymphocyte proliferative responses to phytohaemagglutinin (PHA) stimulation were determined before and after pelvic irradiation with 40 Gy in 10 patients with endometrial carcinoma stage I. The MLC responses were depressed after irradiation, and were not normalized after adherent cell depletion. Adherent cells did not significantly suppress the responses of non-adherent lymphocytes. Indomethacin could not restore the depressed PHA responses occurring after irradiation. Autologous post treatment serum was not suppressive. The results favour the assumption that irradiation causes immunodepression by injuring the responding cells or some amplifier system, rather than by activation of suppressor systems.

Vascular endothelial growth factor (VEGF) and neuroglobin (Ngb) participate in neuronal responses to hypoxia and ischemia, but the relationship between their effects, if any, is unknown. To address this issue, we measured Ngb levels in VEGF-treated mouse cerebrocortical cultures and VEGF levels in cerebrocortical cultures from Ngb-overexpressing transgenic mice. VEGF stimulated Ngb expression in a VEGFR2/Flk1 receptor-dependent manner, whereas Ngb overexpression suppressed expression of VEGF. These findings provide further insight into hypoxia-stimulated neuronal signaling pathways.

Vibrio cholerae O1 in a river water specimen in South Africa was reported, and a public health response followed in order to prevent an outbreak. Further investigation determined this to be a pseudoalert of V. cholerae O1, possibly linked to laboratory contamination. Following culture of bacteria from the water specimen, the testing laboratory possibly contaminated the culture with a V. cholerae O1 reference strain and then mistakenly reported isolation of V. cholerae O1. PMID:22162543

Objectives: To evaluate the implementation of a quality management system based on ISO-15189 in a urine culture unit. Design and methods: The effectiveness of improvement actions was measured by quality indicators. Results: The errors in the pre-analytical phase and the rate of contaminated urine decreased significantly. The traceability, response time and external quality control were fulfilled. Conclusions: The implementation of ISO-15189 was effective in improving the management of a urine culture unit.

Direct and safe manipulation of neurons by external means is an increasingly studied therapeutic modality with the potential to treat many neurological diseases. Anticipating such future applications, we investigated reversible bioeffects of very low dose focused ultrasound on neuronal cell morphology and function in vitro. To test morphological changes, undifferentiated PC12 cells were serum-cultured. The culture plates were placed on an inverted optical microscope. An f/1.1 ultrasound transducer with a water-filled coupling cone was focused on the culture and excited with 30-ms 4.67-MHz 100-kPa pulses. To test functional changes, rat hippocampal slices were cultured and individually transferred to the well of a 60-channel multi electrode array. An f/2.1 ultrasound transducer with a water-filled coupling cone was focused on a culture and excited with 100-?s 4.04-MHz 77-kPa pulses. The culture was stimulated before and after the ultrasonic stimulus with a 100-?s 100-?A biphasic electrical stimulus. Optical microscopy of PC12 cultures under insonification revealed that cells that were clustered near the ultrasound focal region elongated by approximately 2 ?m during insonification and returned to approximately their original shapes following insonification. We conclude that the acoustic radiation force is capable of reversibly deforming cultured cells. In the rat hippocampal cultures, the ultrasonically and electrically evoked responses exhibited similar biphasic waveforms. In addition, robust electrically evoked responses following insonification indicated that the insonified cultures remained viable. We conclude that low-dose ultrasound can stimulate neurons; the mechanism is currently under investigation.

This review describes cross-cultural studies of pitch including intervals, scales, melody, and expectancy, and perception and production of timing and rhythm. Cross-cultural research represents only a small portion of music cognition research yet is essential to i) test the generality of contemporary theories of music cognition; ii) investigate different kinds of musical thought; and iii) increase understanding of the cultural conditions and contexts in which music is experienced. Converging operations from ethology and ethnography to rigorous experimental investigations are needed to record the diversity and richness of the musics, human responses, and contexts. Complementary trans-disciplinary approaches may also minimize bias from a particular ethnocentric view.   

In this study, we analyzed a bacterial community closely associated with Cochlodinium polykrikoides that caused harmful algal blooming in the sea. Filtration using a plankton mesh and percoll gradient centrifugation were performed to eliminate free-living bacteria. Attached bacteria were analyzed by culture-dependent and culture-independent methods. Five culturable bacterial strains were isolated and identified from the C. polykrikoides mixed bacterial community. The isolates belonged to ?-Proteobacteria (Nautella sp., Sagittula sp., and Thalassobius sp.) and ?-Proteobacteria (Alteromonas sp. and Pseudoalteromonas sp.). All of the 5 isolates showed algicidal activity against C. polykrikoides and produced extracellular compounds responsible for algicidal properties after entering the statio...

Purpose - The purpose of this research is to examine Gitga'at First Nation approaches and objectives concerning the use of local biological and cultural resources through the lens of a locally-driven proposal to establish an eco-cultural tourism enterprise. Design/methodology/approach - This project was developed in collaboration with the Gitga'at First Nation and employed a qualitative case study approach. Primary data gathering techniques were active participation, semi-structured interviews, focus group discussions and work with key informants. Findings - Participant responses highlight the interconnectivity and importance of social, ecological and cultural integrity in local economic development. Three major principles for resource use were widely expressed: control and management by G...

During the mixed culture of Lactobacillus hilgardii 5w, a common spoilage wine bacteria and Oenococcus oeni X(2)L, an amensalistic growth response of the malolactic bacteria was produced due to a competition for nitrogenous nutrients, mainly peptides. Arginine was fully consumed and peptide concentration diminished 60% with respect to both pure cultures at the end of exponential growth. Histamine release increased 34% with respect to L. hilgardii single culture. Under the poor nutritional conditions present during winemaking, L. hilgardii could increase histamine production and adversely affect malolactic fermentation conducted by O. oeni and hence the quality of the final product. PMID:20361233

Organ culture of blood vessels provides a useful technique to investigate long-term effects of drugs because tissue architecture and function are well preserved. Various growth factors are responsible for structural and functional changes during vascular diseases. We investigated long-term effects of fetal bovine serum (FBS) which contains such factors on endothelium-dependent relaxation using organ-culture method. Rat isolated mesenteric arteries with endothelium were cultured for 3days without or with 10% FBS (FBS). Acetylcholine- and bradykinin-induced endothelium-dependent relaxations were significantly impaired in FBS, whereas sodium nitroprusside-induced relaxation of endothelium-removed artery was unchanged. Morphological examination revealed that endothelium was intact in FBS. Acet...

Introduction Younger breast cancer survivors often lead extremely busy lives with multiple demands and responsibilities, making them difficult to recruit into clinical trials. African American women are even more difficult to recruit because of additional historical and cultural barriers. In a randomized clinical trial of an intervention, we successfully used culturally informed, population-specific recruitment and retention strategies to engage younger African-American breast cancer survivors. Methods Caucasian and African American breast cancer survivors were recruited from multiple communities and sites. A variety of planned recruitment and retention strategies addressed cultural and population-specific barriers and were guided by three key principals: increasing familiarity with the st...

The direct effects of steroid hormones on the production of immunoglobulins and DNA synthesis by human T and B lymphocytes was evaluated in cultures of peripheral blood mononuclear cells. As detected by a reverse hemolytic plaque assay, the addition of 0.1 mM to 10 nM hydrocortisone to lymphocytes in culture in the absence of other stimulants or mitogens, resulted in the dramatic induction of immunoglobulin production with responses comparable to those seen in similar cultures stimulated with pokeweed mitogen. Steroid-stimulated immunoglobulin production was first seen after 48 h and peaked at 8-10 d of culture. The production of IgG, IgA, and IgM was induced following incubation with steroid. Glucocorticoids, but not estrogens or androgens, were capable of mediating this effect, and only compounds with affinity for the glucocorticoid receptor were active. The induction of immunoglobulin production was dependent on both T cells and monocytes; cultures depleted of either cell type did not produce immunoglobulin when stimulated with glucocorticoid hormones. Proliferation of B cells or T cells could not be detected by (/sup 3/H)thymidine incorporation or total cell recovery from steroid-stimulated cultures, even though such cultures demonstrated marked increases in immunoglobulin production. The mechanism responsible for this functional maturation of B cells to become high rate immunoglobulin producing cells is as yet undefined, although it appears to involve more than merely steroid mediated inactivation of suppressor T cells.

The cultured cells and intact plants of Glycyrrhiza glabra (Fabaceae) produce betulinic acid and oleanane-type triterpene saponins (soyasaponins and glycyrrhizin). To elucidate the regulation of triterpenoid biosynthesis in G. glabra, the cDNA of lupeol synthase, an oxidosqualene cyclase (OSC) responsible for betulinic acid biosynthesis, was cloned, and expression patterns of lupeol synthase and two additional OSCs, ?-amyrin synthase and cycloartenol synthase, were compared. The mRNA expression levels of lupeol synthase and ?-amyrin synthase were consistent with the accumulation of betulinic acid and oleanane-type triterpene saponins, respectively. The transcript of lupeol synthase was highly expressed in the cultured cells and root nodules. The transcript of ?-amyrin synthase, an OSC responsible for oleanane-type triterpene biosynthesis, was highly expressed in the cultured cells, root nodules and germinating seeds, where soyasaponin accumulates, and in the thickened roots where glycyrrhizin accumulates. In the cultured cells, the addition of methyl jasmonate up-regulated ?-amyrin synthase mRNA and soyasaponin biosynthesis, but down-regulated lupeol synthase mRNA. Furthermore, the addition of gibberellin A3 down-regulated ?-amyrin synthase mRNA but not lupeol synthase mRNA in the cultured cells. The mRNA levels of cycloartenol synthase, an additional OSC responsible for sterol biosynthesis, in the intact plant and cultured cells were relatively constant in these experiments.   

This study correlates the potential for somatic embryogenesis in cultured leaf explants of oil palm trees with changes in the expression of hormone-responsive genes. The results show that expression of the putative Aux/IAA gene EgIAA9 is specifically correlated to somatic embryogenesis. A regression model for embryogenesis indicated a predictive R2 value of 63.55?% for this expression marker. Auxin is the plant growth regulator used for induction during the early stages of oil palm tissue culture. Expression of EgIAA9 decreased in response to increasing concentrations of exogenous 2,4-D. EgIAA9 expression profiles in cultured leaf explants from highly embryogenic palms, in embryogenic and non-embryogenic calli, and in response to increasing exogenous 2,4-D, suggested that endogenous auxin ...

This research is located within the call for schools to raise the achievement of students from culturally, linguistically and economically diverse (CLED) backgrounds. It combines quantitative and qualitative research methodologies to compare and contrast preservice teachers' (PSTs) responses to "scenarios" that explore beliefs regarding students from disadvantaged communities. Demographic data is used to determine an innovative Cultural Capital Index, while qualitative responses are coded on a continuum of Cultural Responsiveness to establish and analyse pre-service teachers' beliefs. This paper finds that many PSTs hold deficit attitudes regarding disadvantaged groups of students that may render these teachers unable and subconsciously unwilling to affect a system in need of reform. (Contains 7 notes and 6 figures.)

Abstract in portuguese A biologia reprodutiva de Senna sylvestris foi estudada na Estação Ecológica do Panga (EEP), Uberlândia-MG e no distrito de Macaúbas, Patrocínio-MG. S. sylvestris é um arbusto com altura máxima de 5m e período de floração entre Janeiro e Abril. As flores são hermafroditas, zigomorfas, com corola de cor amarelo intenso, dispostas em inflorescências do tipo panícula. O estigma é do tipo úmido não papiloso e crateriforme, possuindo pêlos ao seu redor. As an (more) teras são poricidas e o androceu é heterântero, com três estaminódios, quatro estames menores na porção superior da flor, dois estames maiores na porção inferior e entre eles um estame central com antera mais delgada. S. sylvestris é auto-estéril e não agamospérmica, com auto-esterilidade envolvendo fenômenos de ação tardia. Tubos polínicos de autopolinizações e polinizações cruzadas crescem e penetram os óvulos, mas frutos e sementes só se desenvolvem após polinizações cruzadas com anteras grandes. Os estames menores e o central de antera mais delgada, produzem pólen inviável. Estes estames servem apenas como pontos de fixação durante as visitas das abelhas. O pólen das anteras pequenas pode formar tubos polínicos, mas nenhum fruto resultou das polinizações com este tipo de pólen. Os principais visitantes e polinizadores são abelhas grandes Xylocopa brasilianorum, Oxaea flavescens, Bombus morio e espécies do gênero Centris que coletam pólen por vibração das anteras. Foram observadas diferenças na riqueza de espécies de abelhas entre as áreas e entre os anos de estudo. Menor número de polinizadores em Macaúbas pode ser explicado pelo maior grau de perturbação da vegetação. Entretanto diferenças na riqueza e no número de polinizadores não parecem explicar a produção natural de frutos, que foi baixa nas espécies estudadas. Abstract in english The reproductive biology of Senna sylvestris was studied in the Panga Ecological Station, Uberlândia and in the Macaúbas district of Patrocínio, Minas Gerais state, Central Brazil. S. sylvestris is a shrub or small tree up to 5m which flowers from January to February. Flowers are hermaphrodite, zigomorphous, with an yellow corolla and disposed in paniculate inflorescences. The stigma is wet, non-papillous and crateriform, surrounded by hairs. Anthers are poricidal in a (more) heteranterous androecium, with three staminodia, four small stamens in the upper portion and two larger ones below with another one with a thin anther in between them. S. sylvestris is self-sterile and non-apomictic, with self-sterility involving late-acting self-incompatibility type phenomena. Self- and cross pollen tubes grow to the ovary and penetrate ovules without any detectable diference, but fruits and seeds developed only from cross pollinations using pollen from the two larger anthers. Smaller stamens and even the large one with a thinner anther produced non-viable pollen. These stamens serve as holding places for the visiting bees. Pollen from these anthers may germinate but no fruit resulted from pollination with this kind of pollen. The main visitors and pollinators were large bees Xylocopa brasilianorum, Oxaea flavescens, Bombus morio, and species of Centris and all collect pollen by vibrating the anthers (buzz pollination). Differences in pollinator species richness were observed between sites and study years. Smaller number of pollinators in Macaúbas may be the result of the increased disturbance in that area. However, species richness and number of pollinators did not explain the constantly low fruit set in the studied species.

The proliferative response to beryllium chloride of cells in a population of human lung fibroblasts was quantitatively assessed using time-lapse cinematography. A dose of 0.02 ..mu..g Be/ml, known to decrease the growth rate of fibroblasts, affects an estimated 75% of the cells in the population, increasing their interdivision time (IDT) by approximately 5 hr. The differences in mean ln(IDT) between treated and control cells were essentially constant for comparable culture sizes ranging from 25 to 250 cells. There was no correlation between mother and daughter cell IDTs in control or treated culture at any culture size. IDTs of sister pairs were highly correlated in control cultures at selected culture sizes while sister pair IDTs of treated cultures were not. The data suggest that while sister pair IDTs of treated cultures were not. The data suggest that while beryllium alters the IDT of fibroblasts, an effect not related to culture size, any given cell affected by beryllium does not impart effects of the mineral to its progeny.

The specific function of the epithelium as critical barrier between the intestinal lumen and the organism's internal microenvironment is reflected by permanent maintenance of intercellular junctions and cellular polarity. The intestinal epithelial cells are responsible for absorption of nutritional components, facing mechanical stress and a changing oxygen supplementation via blood stream. Oxygen itself can regulate the barrier and the absorptive function of the epithelium. Therefore, we compared the dish cell culture, the transwell-like membrane culture and the oxygen enriched air-liquid interface (ALI) culture. We demonstrated strong influence of the different culture conditions on morphology and function of intestinal porcine epithelial cell lines in vitro. ALI culture resulted in a significant increase in cell number, epithelial cell layer thickness and expression as well as apical localisation of the microvilli-associated protein villin. Remarkable similarities regarding the morphological parameters were observed between ALI cultures and intestinal epithelial cells in vivo. Furthermore, the functional analysis of protein uptake and degradation by the epithelial cells demonstrated the necessity of sufficient oxygen supply as achieved in ALI cultures. Our study is the first report providing marked evidence that optimised oxygen supply using ALI cultures directly affects the morphological differentiation and functional properties of intestinal epithelial cells in vitro. PMID:21681518

This paper examines three key considerations underlying the programmatic management of cultural resources that may be affected by a large federal project. These considerations are statutory background and the compliance process, cultural resource compliance tasks, and quality assurance. The first consideration addresses the legal requirements and steps that must be met and taken for federal agencies to fulfill their cultural resource compliance responsibilities. The second consideration focuses on the tasks that must be performed by technical specialists to facilitate related federal and state compliance actions. The third consideration ensures that compliance requirements are being properly fulfilled. In the technical literature and compliance planning, archaeological and historic sites and Native American cultural resources are grouped under the general heading of cultural resources. Also included under this heading are the traditions and resources of Folk societies. Cultural resources encompass both material and nonmaterial aspects of our cultural heritage and include buildings, structures, objects, sites, districts, archaeological resources, places of religious importance, and unique, distinctive, or unusual lifeways. For compliance purposes, it is useful to treat these resources within four roughly chronological culture-historical periods: prehistoric, ethnohistoric, historic, and contemporary. 6 refs., 6 tabs.

This learning package is designed for use in conjunction with inservice training for law enforcement personnel in California as well as for use in law enforcement academies and community college administration of justice classes in police community relations. It consists of a manual on cultural awareness and a companion workbook. Based in part on the categories outlined by the California Commission on Peace Officer Standards and Training, the learning package reflects the broader area of cultural awareness by including more information from cultural anthropology and sociology. The manual contains the objectives and basic information to be covered. Topics include the following: introduction to cultural awareness; legal basis for cultural training; elements of culture; intercultural dynamics; conflict perspective; structural functionalism; interactionist perspective; gender (dynamics); sexual harassment policy; customs; art; music; food; ceremonies and rituals; family and kinship; marriage; religion; dress or appearance; values; bias; language; attitudes toward law enforcement; immigrant (terms); cultural change; immigrant and racial groups; cultural perception; hate crimes; communication skills; and social stratification. The workbook is designed around the use of Gestalt psychology and behavioral psychology with cued exercises and opportunities to use the manual to revise responses. (YLB)

Osteogenic maturation of the osteoblast is crucial for bone formation. In this study, multi-walled carbon nanotubes (MWCNTs) and graphite (GP) were pressed as compacts. The greater ability of carbon nanotubes to adsorb proteins, compared with graphite, was shown. Human osteoblast-like SaoS2 cells were cultured and the cell response to the two kinds of compacts was compared in vitro. Meanwhile, we used cell culture on the culture plate as a control. Assays for osteonectin, osteopontin and osteocalcin gene expression, total protein (TP) amount, alkaline phosphatase activity (ALP) and DNA of cells cultured on the samples were done. During the conventional culture, significantly higher osteonectin, osteopontin and osteocalcin gene expression level, ALP/DNA and TP/DNA on carbon nanotubes were found. To confirm the hypothesis that the larger amount of specific proteins adsorbed on the carbon nanotubes was crucial for this, the compacts were pre-soaked in culture medium having additional recombinant human bone morphogenetic protein-2 (rhBMP-2) before cell culture. Compared with GP, osteonectin, osteopontin and osteocalcin gene expression level, ALP/DNA and TP/DNA of the cells tested increased more on the MWCNTs after the compacts were pre-soaked in the culture medium with rhBMP-2. The results indicated that the carbon nanotubes might induce osteogenic maturation of the osteoblast by adsorbing more specific proteins.

Background and Aims The close relationship between distylic Cordia leucocephala and the bee Ceblurgus longipalpis, both endemic to the Caatinga, north-east Brazil, was investigated, emphasizing reproductive dependence, morphological adaptations of the partners, and pollen flow. Methods In the municipality of Pedra, in the Caatinga of Pernambuco, the breeding system and reproductive success of C. leucocephala, its interaction with flower visitors and inter- and intramorph pollen flow were determined. Key Results The bee Ceblurgus longipalpis, the unique flower visitor and effective pollinator of self-incompatible Cordia leucocephala, presents morphological features adapted to exploit hidden pollen and nectar in the long and narrow corolla tubes. Pollen of low-level anthers is collected with hairs on prolonged mouthparts and pollen of high-level anthers with clypeus, mandibles, and labrum, showing pollen removal from both levels with the same effectiveness. In both morphs, this results in similar legitimate, i.e. intermorph cross-pollen flow. Illegitimate pollen flow to stigmas of pin flowers, however, was much higher than to stigmas of thrum flowers. Moreover, more illegitimate pollen was transported to stigmas of pin and less to those of thrum flowers when compared with legitimate pollen flow. Conclusions The study reveals a one-to-one reproductive inter-dependence between both partners. Data indicate that this relationship between bee species and plant species is one of the rare cases of monolecty among bees. Monotypic Ceblurgus longipalpis, the only rophitine species of Brazil, evolved prolonged mouthparts rare among short-tongued bees that enable them to access pollen from flowers with short-level anthers hidden for bees of other species, and nectar at the base of the flower tube. PMID:17589193

Suspension-cultured cells of Solanum lycopersicum cv Micro-Tom were used to evaluate the effect of methyl jasmonate and cyclodextrins, separately or in combination, on the induction of defense responses. An extracellular accumulation of two sterols (isofucosterol and b-sitosterol) and taraxasterol, a common tomato fruit cuticular triterpene, were observed. Their levels were higher in Micro-Tom tomato suspension cultured cells elicited with cyclodextrins than in control and methyl jasmonate-treated cells. Also, their accumulation profiles during the cell growth phase were markedly different. The most striking feature in response to cyclodextrin treatments was the observed enhancement of taraxasterol accumulation. Likewise, the exogenous application of methyl jasmonate and cyclodextrins indu...

Studies were undertaken to define the helper T-cell requirements for in vitro specific antibody production to influenza virus. Subpopulations of human T cells were separated on the basis of their reactivity with the monoclonal antibody OKT4. B cells cultured with OKT4+ T cells produced specific antibody to influenza virus, while B cells cultured with OKT4- T cells did not. Irradiation (1200 rads) of the OKT4- subset to potentially eliminate suppressor-cell activity did not augment the helper-cell function of that subset. Thus, unlike the cytotoxic T-cell response to influenza, help for an in vitro antibody response is mediated only by OKT4+ T cells.

Internal attribution for bad events, along with stable and global attributions, has been regarded as a component of pessimism, a precursor of negative work outcomes. Most evidence in support of this conceptualisation has come from research conducted in individualist cultures. We questioned if internal attribution has the same pessimistic implication in a collectivist culture. Findings from two studies conducted on Chinese employees supported our expectations that the stability and globality dimensions (but not the internality dimension) would predict disengagement responses (such as quitting and being neglectful at work) and lack of engagement responses (such as voicing suggestions and being loyal to the organisation). A reconceptualisation of pessimism in the workplace is therefore necess...

This study is grounded in the debate surrounding the perceived value of environmental social responsibility (ESR). Applying the Managerial Theory of the Firm, in-depth interviews were conducted to identify managerial motives, perceptions, and perceived value of ESR. Using sport and public assembly facilities as the research context, environmentally responsible information was obtained from facility managers who were members of the International Association of Venue Managers. In total, 15 one-hour, interviews with key facility personnel demonstrate that (1) internal stakeholder pressure, (2) organizational culture, (3) financial cost-benefit, (4) competitiveness, and (5) ethical motives were the drivers for ESR engagement. Taken together, the findings suggest that establishing a culture of ...

PurposeWe determined toll-like receptor expression in normal human urothelium and functional responses in normal human urothelial cell cultures to bacterial lipopolysaccharide via toll-like receptor-4 and to flagellin via toll-like receptor-5. Materials and MethodsToll-like receptor protein expression was examined immunohistochemically. Toll-like receptor transcript expression was determined in freshly isolated urothelium, and in proliferating and differentiated normal human urothelial cultured cells. Lipopolysaccharide binding was assessed by flow cytometry. Functional responses of proliferating and differentiated normal human urothelial cells to lipopolysaccharide and flagellin were determined by interleukin-6 and 8 secretion, and transcription factor activation. Polymyxin B and siRNA we...

The field of migration studies has well-documented children and youths roles as `culture brokers,' by which they mediate relationships, information, and services between the immigrant household and the institutions of the host society. There is growing interest in understanding the contribution of children and youth to socially valued reproductive activity within immigrant households in the United States. Ethnographic studies reveal that children and adolescents in immigrant families have significant responsibilities related to daily life and family functioning. This article focuses on the practice of sibling caretaking, in which older children supervise and socialize younger children, according to culturally informed family roles, responsibilities, and obligations. The purpose of this rev...

Radiation doses of 20, 50 or 100 Gy caused the same time related decrease for RNA and proteoglycan (PG) synthesis in embryonic cartilage in vitro (4 days culture). In this paper, participation of lysosomes in this radiation response is investigated. Therefore, we employ a cytochemical method using beta-glycerophosphate as substrate for acid phosphatase (AP) detection. Increase of AP was found 2 days after irradiation and increased during the whole culture period. The increase was more pronounced with a higher radiation dose. Stimulation of AP activity explains the observed radiation response of RNA and PG synthesis.

Several studies have been conducted to determine the increase of CO2 concentration on the species' growth response. Previous studies revealed that fast-growing species have a greater growth response to high concentrations of CO2 than slow-growing species but it is still unknown if this is a direct response or not. This study examined the relationship between growth rate and response to CO2 and finding which genetic traits enhance response to high CO2 levels. This study was conducted on twenty-nine genotypes of Picea glauca which were cultured in pots at ambient and elevated CO2 and average relative growth rate (RGR) was determine all along the experiment. Results showed no sign of relationship between RGR and CO2 growth response and differences in RGR were linked to several different traits. This study highlighted that more researches are required to determine which traits are responsible for differences in CO2 growth responses both within and between species.

An in vitro system using neuroglia from neonatal rat brain was developed to examining the morphologic, immunocytochemical and biochemical response of oligodendroglia to ionizing radiation. Following acute [gamma]-radiation at day-in-culture (DIC) 8, oligodendrocyte counts at DIC 14 were 55% to 65% of control values after 2 Gy, and 29% to 36% after 5 Gy. Counts increased to near-normal levels at DIC 21 in the 2 Gy group and to 75% of normal in the 5 Gy group. Myelin basic protein levels (MBP) at DIC 14 were 60% of control values after 2 Gy, and 40% after 5 Gy. At DIC 21, MBP after 2 Gy was 45% greater than that observed at DIC 14, but MBP, as a fraction of age-matched control values, dropped from 60% to 50%. Following 5 Gy, absolute MBP changed little between DIC 14 and DIC 21, but decreased from 40% to 25% of control cultures. It was concluded that oligodendrocytes in irradiated cultures had significantly lower functional capacity than did unirradiated controls. The response to split-dose irradiation indicated that nearly all sublethal damage in the oligodendrocyte population (and its precursors) was repaired within 3 h to 4 h. At DIC 14, the group irradiated in a single fraction had significantly lower oligodendrocyte counts than any group given split doses; all irradiated cultures had marked depression of MBP synthesis, but to significant differences referable to time interval between doses. At DIC 21, cultures irradiated at intervals of 0 h to 2 h had similar oligodendrocyte counts to one another, but these counts were significantly lower than in cultures irradiated at intervals of 4 h to 6 h; MBP levels remained depressed at DIC 21 for all irradiated cultures. The oligodendrocyte response to dose rate (0.03 to 1.97 Gy/min) was evaluated at DIC 14 and DIC 21. Exposure at 0.03 Gy/min suppressed oligodendrocyte counts at DIC 21 less than did higher dose rates in 5-Gy irradiated cultures.

Cassia siamea plants growing at two different sites (polluted and non-polluted) on two important roads of Agra city exhibited significant differences in their flowering phenology and floral morphology. The flowering in plants growing at polluted site is delayed and there was a marked reduction in flowering density, flowering period, size of floral parts, pollen fertility, fruit and seed-set. SEM observations revealed the presence of well developed glandular structures and reduction in the number and size of large stomata on the anther surface at polluted site. These changes were found to be closely associated with the extent of air pollution caused mainly by significant in the number of automobiles. PMID:15847337

The reproductive morphology of Sargentodoxa cuneata (Oliv) Rehd. et Wils. is investigated through field, herbarium, and laboratory observations. Sargentodoxa may be either dioecious or monoecious. The functionally unisexual flowers are morphologically bisexual, at least developmentally. The anther is tetrasporangiate, and its wall, of which the development follows the basic type, is composed of an epidermis, endothecium, two middle layers, and a tapetum. The tapetum is of the glandular type. Microspore cytokinesis is simultaneous, and the microspore tetrads are tetrahedral. Pollen grains are two-celled when shed. The mature ovule is crassinucellate and bitegmic, and the micropyle is formed only by the inner integument. Megasporocytes undergo meiosis resulting in the formation of four megas...

  We did in situ hybridization to identify the sites at which DnaJ homologs (SGJ1 and SGJ3) and Hsp70 were expressed in male and female flowers of the Japanese willow, Salix gilgiana Seemen. Transcripts of SGJ1, SGJ3, and Hsp70 were detected in microspores and in the tapetal layers of immature anthers, in the inner integument of ovules, and in the funicular epidermal layers of ovaries. Transcripts of SGJ3 and Hsp70 were also detected in the nectaries of male and female flowers, in which transcripts of SGJ1 were undetectable.   

Pollination biology of Annona squamosa was investigated in the context of functional specialization and pollination syndrome. Hermaphroditic flower exhibited protogynous dichogamy as stigma became receptive a day before anther dehiscence. Flowers produce moderately high number (16,280+/-324) of spherical and medium size (98.6mmx87.3mm) pollen with pollen/ovule ratio of 120.6. Nitidulid beetles; Carpophilos domidiatus and Carpophilos hemipterous were identified as pollinators in terms of visitation frequency, pollination potential index score and pollination efficiency. However C. hemipterous was relatively effective pollinator than C. domidiatus. Floral traits and specific reward system of A. squamosa clearly exhibit obligate specialization by filtering only C. hemipterous and C. domidiatu...

Morphological and cytological studies are complementary approaches to understand the molecular mechanisms that regulate floral developmental pathways. To better understand abnormal mutant phenotypes in floral development, we conducted detailed observations and investigations of the morphology, cytology, and cell ultrastructure of wild-type Chinese cabbage (Brassica campestris L. ssp. chinensis Makino and syn. B. rapa ssp. chinensis) flowers when they developed from primordia to anthesis. First, we measured bud and organ length with a stereo microscope and observed the developmental status and characteristics of the floral organs using a scanning electron microscope; then we made thin slices of anthers to observe the developmental stage and characteristics of pollen using an optical microsc...

Background and Aims The close relationship between distylic Cordia leucocephala and the bee Ceblurgus longipalpis, both endemic to the Caatinga, north-east Brazil, was investigated, emphasizing reproductive dependence, morphological adaptations of the partners, and pollen flow. Methods In the municipality of Pedra, in the Caatinga of Pernambuco, the breeding system and reproductive success of C. leucocephala, its interaction with flower visitors and inter- and intramorph pollen flow were determined. Key Results The bee Ceblurgus longipalpis, the unique flower visitor and effective pollinator of self-incompatible Cordia leucocephala, presents morphological features adapted to exploit hidden pollen and nectar in the long and narrow corolla tubes. Pollen of low-level anthers is collected with...

Background: The opportunistic pathogen, Staphylococcus aureus, experiences fluctuations in fluid shear during infection and colonization of a human host. Colonization frequently occurs at mucus membrane sites such as in the gastrointestinal tract where the bacterium may experience low levels of fluid shear. The response of S. aureus to low fluid shear remains unclear. Methods: S. aureus was cultured to stationary phase using Rotating-Wall Vessel (RWV) bioreactors which produce a physiologically relevant low fluid shear environment. The bacterial aggregates that developed in the RWV were evaluated by electron microscopy as well as for antibiotic resistance and other virulence-associated stressors. Genetic expression profiles for the low-shear cultured S. aureus were determined by microarray analysis and quantitative real-time PCR. Results: Planktonic S. aureus cultures in the low-shear environment formed aggregates completely encased in high amounts of extracellular polymeric substances. In addition, these aggregates demonstrated increased antibiotic resistance indicating attachment-independent biofilm formation. Carotenoid production in the low-shear cultured S. aureus was significantly decreased, and these cultures displayed an increased susceptibility to oxidative stress and killing by whole blood. The hfq gene, associated with low-shear growth in Gram negative organisms, was also found to be down-regulated in S. aureus. Conclusions: Collectively, this data suggests that S. aureus decreases virulence characteristics in favor of a biofilm-dwelling colonization phenotype in response to a low fluid shear environment. Furthermore, the identification of an Hfq response to low-shear culture in S. aureus, in addition to the previously reported responses in Gram negative organisms, strongly suggests an evolutionarily conserved response to mechanical stimuli among structurally diverse prokaryotes.

The aim of this study was to standardize in vitro culture conditions for human acute lymphoblastic leukemia (ALL) cells. The cells were cultured in medium containing 10% fetal calf serum (FCS) and in the four serum-free media X-vivo 10, X-vivo 15, X-vivo 20 and Stem Span. Native ALL blasts could proliferate in all four serum-free media, but the strongest responses were usually observed with Stem Span. Native leukemia blasts were also cultured in the presence of various single cytokines or cytokine combinations. The highest proliferation was usually observed in the presence of Flt3-Ligand (Flt3-L) when single cytokines were examined, and these responses could be further increased especially by combining Flt3-L with interleukin 3 (IL3), IL7 or stem cell factor (SCF). Proliferation could also be increased when ALL blasts were cultured in the presence of two commercially available fibroblast cell lines (Hs27 and HFL1). Based on these results we suggest that in vitro culture conditions for native human ALL blasts can be standardized by using serum-free culture media supplemented with exogenous Flt3-L+IL3+SCF, and the use of accessory cells can also be standardized by using well-characterized fibroblast cell lines. Detectable ALL blast proliferation can then be observed for most patients. Our experimental model can thereby be used for in vitro evaluation of possible antileukemic treatment strategies, and it will then allow comparison of experimental results between different studies. PMID:12620297

Cultural Content Technology(CT, Culture Technology) for the development of cultural industry and the commercialization of technology, cultural contents, media, mount, pass the value chain process and increase the added value of cultural products that are good for all forms of intangible technology. In the field of Culture Technology, Music by analyzing the characteristics of the development of a variety of applications has been studied. Associated with EEG measures and the results of their research in response to musical stimuli are used to detect and study is getting attention. In this paper, the musical stimuli in EEG signals by amplifying the corresponding reaction to the averaging method, ERP (Event-Related Potentials) experiments based on the process of extracting sound methods for removing noise from the ICA algorithm to extract the tone and noise removal according to the results are applied to analyze the characteristics of EEG. In addition, using LORETA(Low Resolution Brain Electromagnetic Tomography)program have attempt to simulate the brain CT scan.LORETA programs by using the active region of the brain to review and listen to music when trying to identify new areas of the brain response that is meaningful.   

Prediction of human response to potential therapeutic drugs is through conventional methods of in vitro cell culture assays and expensive in vivo animal testing. Alternatives to animal testing require sophisticated in vitro model systems that must replicate in vivo like function for reliable testing applications. Advancements in biomaterials have enabled the development of three-dimensional (3D) cell encapsulated hydrogels as in vitro drug screening tissue model systems. In this study, we have developed an in vitro platform to enable high density 3D culture of liver cells combined with a monolayer growth of target breast cancer cell line (MCF-7) in a static environment as a representative example of screening drug compounds for hepatotoxicity and drug efficacy. Alginate hydrogels encapsulated with serial cell densities of HepG2 cells (10{sup 5}-10{sup 8} cells/ml) are supported by a porous poly-carbonate disc platform and co-cultured with MCF-7 cells within standard cell culture plates during a 3 day study period. The clearance rates of drug transformation by HepG2 cells are measured using a coumarin based pro-drug. The platform was used to test for HepG2 cytotoxicity 50% (CT{sub 50}) using commercially available drugs which further correlated well with published in vivo LD{sub 50} values. The developed test platform allowed us to evaluate drug dose concentrations to predict hepatotoxicity and its effect on the target cells. The in vitro 3D co-culture platform provides a scalable and flexible approach to test multiple-cell types in a hybrid setting within standard cell culture plates which may open up novel 3D in vitro culture techniques to screen new chemical entity compounds. - Graphical abstract: Display Omitted Highlights: > A porous support disc design to support the culture of desired cells in 3D hydrogels. > Demonstrated the co-culture of two cell types within standard cell-culture plates. > A scalable, low cost approach to toxicity screening involving multiple cell types.

Recent studies have revealed the existence of stem cells in various human tissues including dental structures. We aimed to establish primary cell cultures from human dental pulp and periodontal ligament, to identify multipotential adult stem cells in these cultures, and to study the differentiation capacity of these cells to osteogenic and to neuronal fates. Dental pulp and the periodontal ligament were isolated from extracted human wisdom teeth. The extracellular matrix was enzymatically degraded to obtain isolated cells for culturing. Both dental pulp and periodontal ligament derived cultures showed high proliferative capacity and contained a cell population expressing the STRO-1 mesenchymal stem cell marker. Osteogenic induction by pharmacological stimulation resulted in mineralized differentiation as shown by Alizarin red staining in both cultures. When already described standard neurodifferentiation protocols were used, cultures exhibited only transient neurodifferentiation followed by either redifferentiation into a fibroblast-like phenotype or massive cell death. Our new three-step neurodifferentiation protocol consisting of (1) epigenetic reprogramming, then (2) simultaneous PKC/PKA activation, followed by (3) incubation in a neurotrophic medium resulted in robust neurodifferentiation in both pulp and periodontal ligament cultures shown by cell morphology, immunocytochemistry and real time PCR for vimentin and neuron-specific enolase. In conclusion, we report the isolation, culture and characterization of stem cell containing cultures from both human dental pulp and periodontal ligament. Furthermore, our data clearly show that both cultures differentiate into mineralized cells or to a neuronal fate in response to appropriate pharmacological stimuli. Therefore, these cells have high potential to serve as resources for tissue engineering not only for dental or bone reconstruction, but also for neuroregenerative treatments. PMID:20388961

Cultured human peripheral blood mononuclear cells suppressed the allogeneic response of fresh autologous lymphocytes. This suppressor activity developed gradually over a period of one week. The cells primarily responsible for this effect were enriched by Ficoll density gradient centrifugation. It was found that the suppressor cell is a large, low density nylon wool adherent, radioresistant, phagocytic, and nonspecific esterase positive mononuclear cell. Moreover, these cells did not form E rosettes and were Fc positive. Electron microscopy confirmed that suppressor cells were macrophage like. Suppressor activity was not due to cytotoxicity, crowding, or steric hinderance by the cultured cells. The suppressor macrophage population did not appear to inhibit the allogeneic response via prostaglandin or arginase release, or interfere with the tritiated thymidine uptake by release of endogenous thymidine. The above system is viewed as an in vitro model of immune regulation by suppressor macrophages, in the context of allogeneic response.

The purpose of this project is improvement of investigation, materialization and evaluation techniques on effectiveness for functional natural compounds throughout development of tissue/cell culture techniques for mass production of useful metabolites using radiation. Research scope includes 1) Development of a technique for radiation tissue and cell culture, 2) Database construction for radiation response in plants and radiation effects, 3) Construction of general-purpose national based techniques of cell culture technique using radiation. Main results are as follow: mass culture of the adventitious roots of mountain ginseng (Panax ginseng C. A. Meyer) roots using rare earth elements in bioreactor: characterization of a transcription factor EoP gene from centipedegrass and the transcription regulation of LexA from Synechocystis sp PCC6803 and E. coli: identification of gamma-ray induced hydrogenase synthesis in hox gene transformed E. coli: transformation and the selection of the EoP transgene from Arabidopsis, rice and lettuce: Identification of the maysin and maysin derivatives in centipedegrass: characterization of gamma-ray induced color change in Taxus cuspidata: verification of the expression of antioxidant proteins (POD, APX and CAT) to gamma-ray in Arabidopsis: comparison of the response of the expression level to gamma-ray or H{sub 2}O{sub 2} in Arabidopsis; verification of the responses and effects to gamma-ray from plants (analysis of NPQ and ROS levels): the development method for rapidly enhancing maysin content of centipede grass; establishment of mass culture system for red beet

The photodynamic toxicity of benzo(a)pyrene (BaP) to Paramecium caudatum and a variety of other in vivo systems is well established. Recent improvements in the culture of P. caudatum have facilitated standardization of the photodynamic response. A simple bioassay procedure, based on this response, was developed as a preliminary to the assay for photodynamic toxicity of crude organic extracts, such as atmospheric pollutants.

The aim of this article is to explore explanations for unethical healthcare practice and identify educational responses. The meaning of unethical practice is outlined and causes of it are suggested, primarily relating to individual perpetrators and organisational culture or climate. Empirical and theoretical literature is reviewed and research findings are discussed. Individual resilience and the ethical climate of healthcare organisations are considered as responses to unethical practice. Role modelling is explored, acknowledging the role of effective leadership. PMID:22848954

Gentians (Gentiana triflora, G. scabra, and hybrids of the two) are mainly cultivated as ornamental flowers in Japan. Because gentians are allogamous plants, their diversity and heterozygosity have become a major problem. Recently, explants were clonally cultured to maintain genetic purity, but culture conditions have not been studied systematically, thus the essential nutrients required for gentian culture are unknown. We therefore investigated the effects of potassium (K) and phosphorus (P) deficiency in culture media. Explants grew under K or P deficiency conditions, but P deficiency caused the formation of new structures which are similar to overwintering buds. To elucidate the mechanism behind the gentian response to mineral deficiency, we performed targeted metabolome analyses using ...

The addition of 2, 4-dichlorophenoxyacetic acid (2, 4-D) to Vitis sp. cell cultures significantly enhanced the production of quercetin 3, 7, 4?-tri-O-glucoside, 3, 7-di-O-glucoside and 3, 4?-di-O-glucoside from quercetin. This enhancement of glucosylation by 2, 4-D was also observed in cell cultures of other plant species. The activity of UDP-glucose:flavonol glucosyltransferase (UFGT) in cell-free extracts of Vitis sp. cell cultures increased approximately 10-fold, 48h after the addition of 2, 4-D to the culture medium. The UFGT activity increased linearly up to 15h and showed a maximal response to the addition of 10-50mg/l of 2, 4-D at 48h. The promotive effect of 2, 4-D was inhibited by cycloheximide suggesting that de novo protein synthesis was involved in this phenomenon. Interestingly, similar promotive effects on the UFGT activity were observed for other phytohormones such as kinetin and several anti-auxins.   

An efficient protocol was developed for Agrobacterium tumefaciens-mediated transformation of tomato (Solanum lycopersicum) cultivars using cotyledon explants. The transformation frequency was assessed in response to several different factors, including seed germination medium, seedling age, pre-culture duration, pre-culture and co-cultivation media, inoculation medium, medium pH, washing medium, and kanamycin concentration in initial selection medium. Cotyledons excised from 6-d-old seedlings germinated on half-strength Murashige and Skoog?s (MS) basal medium containing 8.9 ?M benzyladenine (BA) produced the most suitable explant material. Six days of explant pre-culture and 5 min inoculation with Agrobacterium culture in MS medium, containing 8.9 ?M BA, 9.3 ?M kinetin, and 0.4 mg l?1 thia...

Moving from cell monolayers to three-dimensional (3D) cultures is motivated by the need to work with cellular models that mimic the functions of living tissues. Essential cellular functions that are present in tissues are missed by 'petri dish'-based cell cultures. This limits their potential to predict the cellular responses of real organisms. However, establishing 3D cultures as a mainstream approach requires the development of standard protocols, new cell lines and quantitative analysis methods, which include well-suited three-dimensional imaging techniques. We believe that 3D cultures will have a strong impact on drug screening and will also decrease the use of laboratory animals, for example, in the context of toxicity assays.

Cyclodextrin glucanotransferase production from Bacillus clausii E16, a new bacteria isolated from Brazilian soil samples was optimized in shake-flask cultures. A 24 full-factorial central composite design was performed to optimize the culture conditions, using a response surface methodology. The combined effect among the soluble starch concentration, the peptone concentration, the yeast extract concentration, and the initial pH value of the culture medium was investigated. The optimum concentrations of the components, determined by a 24 full-factorial central composite design, were 13.4 g/L soluble starch, 4.9 g/L peptone, 5.9 g/L yeast extract, and initial pH 10.1. Under these optimized conditions, the maximum cyclodextrin glucanotransferase activity was. 5.9 U/mL after a 48-h fermentation. This yield was 68% higher than that obtained when the microorganism was cultivated in basal culture medium.

Abstract Accumulation of secondary metabolites is one of the common reactions of plants to ozone exposure in nature. To investigate the effect of ozone on the production of desired compounds of plant cell cultures, we assayed hypericin production of Hypericum perforatum suspension cell cultures treated with different doses of ozone at different culture phases. The results show that hypericin contents of the cells treated with 60 to 180 nL L-1 ozone are significantly higher than those of the control, showing that ozone exposure may stimulate hypericin synthesis. Hypericin production of the cells treated with ozone at exponential phase is higher than that of lag and stationary phase, which suggests that exponential phase cell cultures are more responsive to ozone exposure than lag and statio...

Cultural sensitivity is recommended in education, but to our knowledge it has not been defined operationally. We found no research suggesting that behavioral interventions work differently with students differing in ethnicity, gender, or religion, although socialization may play a role in shaping behavior, selecting or responding to particular reinforcers, and judging the acceptability of interventions. Differences in response to behavioral interventions based on cultural identification may exist, but we found no evidence suggesting that behavioral practices are more likely to be effective if they are based on the cultural markers we examined. For behavioral educators, cultural sensitivity requires respect for the individual student based on scientific evidence--measuring the effects of scientific interventions in achieving socially valid behavioral objectives for individual students and their parents.

Understanding Corporate Social Responsibility (CSR) as having explicit policies and implicit norms situated in cultural systems highlights the connections between institutional and cultural structures of nation states and business' commitment to CSR as reflected in the strategies used to communicate CSR to public audiences via the Internet.  To frame CSR from a situated perspective (Matten & Moon 2005) implies a shift in understanding relations between corporations and their stakeholders from a corporate-centered model to a cultural systems perspective.  This paper describes an approach to cultural systems in which can be used to frame our understanding of implicit norms with respect to CSR, and demonstrates how these norms result in different practices of communicating CSR in the WalMart and Maersk corporate websites.

Purpose of the Study: In recent years, there has been an increasing focus on the role of safety culture in preventing costly adverse events, such as medication errors and falls, among nursing home residents. However, little is known regarding critical organizational determinants of a positive safety culture in nursing homes. The aim of this study was to identify organizational climate predictors of specific aspects of the staff-rated resident safety culture (RSC) in a sample of nursing homes. Design and Methods: Staff at 4 Michigan nursing homes responded to a self-administered questionnaire measuring organizational climate and RSC. Multiple regression analyses were used to identify organizational climate factors that predicted the safety culture dimensions nonpunitive response to mistakes...

The present work was conducted to develop a simple and efficient protocol for in vitro shoot regeneration and plantlet formation from unpollinated ovary culture in Allium chinense. Ovaries that were excised from flowers 2-3 days before anthesis and cultured on induction medium with 9.05mM 2,4-dichlorophenoxyacetic acid (2,4-D) and 8.88mM benzylaminopurine (BAP) for 2 weeks gave the best organogenic response (56.32%). For shoot regeneration, the highest percent organogenic ovaries (64.53%) were obtained on B5 induction medium containing 9.05mM 2,4-D and 8.88mM BAP. Ovaries cultured on B5 containing 10.74mM naphtaleneacetic acid (NAA) and 0.44mM BAP exhibited maximum shoot regeneration (60.32%). Shoots elongated and produced normal plantlets when cultured on B5 medium supplemented with 10.74...

An enterprise culture is one in which -certain enterprising qualities - such as self reliance, personal responsibility, boldness and a willingness to take risks in the pursuit of goals - are regarded as human virtues and promoted as such-. Work is not seen as a constraint upon the individual but rather as an opportunity for self-fulfilment. The pervasiveness of enterprise culture has led to observations that it influences aspects of social life beyond the workplace. However, the kind of self that is imagined in enterprise culture is also a male self, with female entrepreneurship measured against a male norm. In this article, then, we present evidence of metaphor as a strategy for coping with the gendered expectations embodied by life in enterprise culture. We show how metaphor is simultane...