Dynamics and elasticity of the fibronectin matrix in living cell culture visualized by fibronectin–green fluorescent protein

Fibronectin (FN) forms the primitive fibrillar matrix in both embryos and healing wounds. To study the matrix in living cell cultures, we have constructed a cell line that secretes FN molecules chimeric with green fluorescent protein. These FN–green fluorescent protein molecules were assembled into a typical matrix that was easily visualized by fluorescence over periods of several hours. FN fibrils remained mostly straight, and they were seen to extend and contract to accommodate movements of the cells, indicating that they are elastic. When fibrils were broken or detached from cells, they contracted to less than one-fourth of their extended length, demonstrating that they are highly stretched in the living culture. Previous work from other laboratories has suggested that cryptic sites for FN assembly may be exposed by tension on FN. Our results show directly that FN matrix fibrils are not only under tension but are also highly stretched. This stretched state of FN is an obvious candidate for exposing the cryptic assembly sites.

Opposite Effects on Spodoptera littoralis Larvae of High Expression Level of a Trypsin Proteinase Inhibitor in Transgenic Plants1

This work illustrates potential adverse effects linked with the expression of proteinase inhibitor (PI) in plants used as a strategy to enhance pest resistance. Tobacco (Nicotiana tabacum L. cv Xanthi) and Arabidopsis [Heynh.] ecotype Wassilewskija) transgenic plants expressing the mustard trypsin PI 2 (MTI-2) at different levels were obtained. First-instar larvae of the Egyptian cotton worm (Spodoptera littoralis Boisd.) were fed on detached leaves of these plants. The high level of MTI-2 expression in leaves had deleterious effects on larvae, causing mortality and decreasing mean larval weight, and was correlated with a decrease in the leaf surface eaten. However, larvae fed leaves from plants expressing MTI-2 at the low expression level did not show increased mortality, but a net gain in weight and a faster development compared with control larvae. The low MTI-2 expression level also resulted in increased leaf damage. These observations are correlated with the differential expression of digestive proteinases in the larval gut; overexpression of existing proteinases on low-MTI-2-expression level plants and induction of new proteinases on high-MTI-2-expression level plants. These results emphasize the critical need for the development of a PI-based defense strategy for plants obtaining the appropriate PI-expression level relative to the pest's sensitivity threshold to that PI.

The Boron Requirement and Cell Wall Properties of Growing and Stationary Suspension-Cultured Chenopodium album L. Cells1

Suspension-cultured Chenopodium album L. cells are capable of continuous, long-term growth on a boron-deficient medium. Compared with cultures grown with boron, these cultures contained more enlarged and detached cells, had increased turbidity due to the rupture of a small number of cells, and contained cells with an increased cell wall pore size. These characteristics were reversed by the addition of boric acid (≥7 μm) to the boron-deficient cells. C. album cells grown in the presence of 100 μm boric acid entered the stationary phase when they were not subcultured, and remained viable for at least 3 weeks. The transition from the growth phase to the stationary phase was accompanied by a decrease in the wall pore size. Cells grown without boric acid or with 7 μm boric acid were not able to reduce their wall pore size at the transition to the stationary phase. These cells could not be kept viable in the stationary phase, because they continued to expand and died as a result of wall rupture. The addition of 100 μm boric acid prevented wall rupture and the wall pore size was reduced to normal values. We conclude that boron is required to maintain the normal pore structure of the wall matrix and to mechanically stabilize the wall at growth termination.

Effects of Xylem pH on Transpiration from Wild-Type and flacca Tomato Leaves1 : A Vital Role for Abscisic Acid in Preventing Excessive Water Loss Even from Well-Watered Plants

The pH of xylem sap from tomato (Lycopersicon esculentum) plants increased from pH 5.0 to 8.0 as the soil dried. Detached wild-type but not flacca leaves exhibited reduced transpiration rates when the artificial xylem sap (AS) pH was increased. When a well-watered concentration of abscisic acid (0.03 μm) was provided in the AS, the wild-type transpirational response to pH was restored to flacca leaves. Transpiration from flacca but not from wild-type leaves actually increased in some cases when the pH of the AS was increased from 6.75 to 7.75, demonstrating an absolute requirement for abscisic acid in preventing stomatal opening and excessive water loss from plants growing in many different environments.

Cellular adherence elicits ligand-independent activation of the Met cell-surface receptor.

Cell adhesion has a fundamental role in the proliferation and motility of normal cells and the metastasis of tumor cells. To identify signaling pathways activated by the adherence of tumor cells, we analyzed the tyrosine phosphorylation of proteins in mouse melanoma cells before and after attachment to substrata. We discovered that cellular adherence activated the protein-tyrosine kinase of the cell surface receptor Met, whose ligand is hepatocyte growth factor and scatter factor. The activation was exceedingly prompt, affected the great majority of Met in the cells, persisted so long as the cells remained adherent, and was rapidly reversed as soon as the cells were detached from substrata. Activation of Met required that cells be adherent but not that they spread on the substratum, and it occurred in the absence of any apparent ligand for the receptor. Ligand-independent activation of Met occurred in several varieties of tumor cells but not in normal endothelial cells that express the receptor. The activation of Met described here may represent a means by which cells respond to mechanical as opposed to biochemical stimuli.

A mammalian 2-5A system functions as an antiviral pathway in transgenic plants.

Resistance to virus infections in higher vertebrates is mediated in part through catalysis of RNA decay by the, interferon-regulated 2-5A system. A functional 2-5A system requires two enzymes, a 2-5A synthetase that produces 5'-phosphorylated, 2',5'-linked oligoadenylates (2-5A) in response to double-stranded RNA, and the 2-5A-dependent RNase L. We have coexpressed these human enzymes in transgenic tobacco plants by using a single plasmid containing the cDNAs for both human RNase L and a low molecular weight form of human 2-5A synthetase under control of different, constitutive promoters. Expression of the human cDNAs in the transgenic plants was demonstrated from Northern blots, by specific enzyme assays, and by immunodetection (for RNase L). Infection of leaves, detached or in planta, of the coexpressing transgenic plants by tobacco mosaic virus, alfalfa [correction of alfafa] mosaic virus, or tobacco etch virus resulted in necrotic lesions. In contrast, leaves expressing 2-5A synthetase or RNase L alone and leaves containing the plasmid vector alone produced typical systemic infections. While alfalfa mosaic virus produced lesions only in the inoculated leaves regardless of the concentration of virus in the inoculum, high, but not low, levels of tobacco etch virus inoculum resulted in escape of virus to uninoculated leaves. Nevertheless, there was a substantial reduction of tobacco etch virus yield as measured by ELISA assay in the coexpressing transgenic plants. These results indicate that expression of a mammalian 2-5A system in plants provides resistance to virus infections.

A membrane-associated form of sucrose synthase and its potential role in synthesis of cellulose and callose in plants.

Sucrose synthase (SuSy; EC 2.4.1.13; sucrose + UDP reversible UDPglucose + fructose) has always been studied as a cytoplasmic enzyme in plant cells where it serves to degrade sucrose and provide carbon for respiration and synthesis of cell wall polysaccharides and starch. We report here that at least half of the total SuSy of developing cotton fibers (Gossypium hirsutum) is tightly associated with the plasma membrane. Therefore, this form of SuSy might serve to channel carbon directly from sucrose to cellulose and/or callose synthases in the plasma membrane. By using detached and permeabilized cotton fibers, we show that carbon from sucrose can be converted at high rates to both cellulose and callose. Synthesis of cellulose or callose is favored by addition of EGTA or calcium and cellobiose, respectively. These findings contrast with the traditional observation that when UDPglucose is used as substrate in vitro, callose is the major product synthesized. Immunolocalization studies show that SuSy can be localized at the fiber surface in patterns consistent with the deposition of cellulose or callose. Thus, these results support a model in which SuSy exists in a complex with the beta-glucan synthases and serves to channel carbon from sucrose to glucan.

Adaptation of Spodoptera exigua larvae to plant proteinase inhibitors by induction of gut proteinase activity insensitive to inhibition.

Tobacco plants were transformed with a cDNA clone of chymotrypsin/trypsin-specific potato proteinase inhibitor II (PI2) under the control of a constitutive promoter. Although considerable levels of transgene expression could be demonstrated, the growth of Spodoptera exigua larvae fed with detached leaves of PI2-expressing plants was not affected. Analysis of the composition of tryptic gut activity demonstrated that only 18% of the proteinase activity of insects reared on these transgenic plants was sensitive to inhibition by PI2, whereas 78% was sensitive in insects reared on control plants. Larvae had compensated for this loss of tryptic activity by a 2.5-fold induction of new activity that was insensitive to inhibition by PI2. PI2-insensitive proteolytic activity was also induced in response to endogenous proteinase inhibitors of tobacco; therefore, induction of such proteinase activity may represent the mechanism by which insects that feed on plants overcome plant proteinase inhibitor defense.

Contemplative Education CenterStage: Training the Mindful Performer

In the late 20th century and early 21st century, contemplative education/studies courses, concentrations, and initiatives have emerged in the academy. Although there has been significant discussion of postsecondary courses and programs that have integrated contemplative views and practices in the literature, there have been few studies of contemplative curricula and pedagogy in higher education. Additionally, there have been even fewer inquiries of the influence of contemplative education on performing arts training within conservatories and college and university departments. The aim of this qualitative study was two-fold: (1) to describe, interpret, and appraise the impact of contemplative education on the curricula of an interdisciplinary conservatory level performing arts program, MFA Contemporary Performance, at Naropa University; and (2) to disclose, compare, and analyze MFA student perceptions of the influence of contemplative education on their professional and personal development. The following questions guided this study: (1) How do faculty and students characterize contemplative education within the MFA in Theater: Contemporary Performance Program? (2) How does contemplative education impact the intended and operational curricula of courses within the MFA Contemporary Performance Program? (3) How do graduate students perceive the effects of contemplative education, offered by the MFA Contemporary Performance Program, on the development of their communication abilities, presence-in-performance, sociolinguistic perspectives, and aesthetic perspectives? Based on the research methodology of educational criticism and connoisseurship, this investigation provides a vivid description and interpretation of the intended and operational curricula of three core courses within the MFA program. These curricula were examined through five dimensions: intentional, curricular, pedagogical, structural, and evaluative. In order to shape our understanding of the contemplative and performative nature of the curricula, the significant and subtle qualities of the courses were further captured by preparation, context-building, reflective, showing, and closing conventions. Since the courses were grounded in postmodern view, they were evaluated according to Doll's criteria of richness, recursion, relations, and rigor for the evaluation of postmodern curricula. MFA first- and second-year students primarily characterized contemplative education as body/mind training for performance and personal development, sitting meditation, and cultivation of mindfulness and awareness. Student perceptions of the impact of contemplative education on the development of their communication abilities, presence-in-performance, sociolinguistic perspectives, and aesthetic perspectives, throughout the course of their two-year training, are presented in a dimensional analysis. The research reveals eight different themes that intersect the three core curricula and interviews with MFA students and faculty. These thematics include inclusivity, nowness, silence, improvisation, goodness, heart, training, and space. The beginning letter of each theme combines to form the acronym, insights. The framework of insights connects and illuminates the most potent aspects of MFA Contemporary Performance values and training.

Modified Dialectical Behavior Therapy Program Applied Within a Time-Limited Corrections Setting

Dialectical Behavioral Therapy (DBT) is an empirically supported therapy developed to treat individuals with Borderline Personality Disorder that has sustained efficacy following completion of the treatment (Linehan, 1993; Van Den Bosch et al., 2005). The core concepts of DBT include mindfulness, interpersonal effectiveness, emotional regulation, and distress tolerance, which seek to foster more functional ways of interacting with others, coping with distress, and managing difficult emotions. Using a standard DBT format in a corrections setting can be difficult due to the population's multifaceted composition. The Denver County Jail is a unique corrections setting because it contains a unit specifically developed for male inmates with mental health issues. A corrections modified, time-limited DBT curriculum was developed to fit the needs of this unique population. During the course of the group, staff appeared to be accepting of the group material and initial feedback from inmates and officers was positive.

Convention, Tradition, and the New Wave : Assessing Clinician Identity within Behavior Therapy

In the first wave, behaviorists questioned the conventional wisdom that inner experience was relevant to understanding human behavior. In the 1970s, cognitive-behavioral theories emphasized the importance of the cognitive element, not just the environment, in explaining and modifying behavior. The third wave is drawn from advances in basic and applied behavior analysis of language, Eastern mystical traditions, and less empirically oriented therapeutic approaches. Examples include Acceptance and Commitment Therapy (ACT), Dialectical Behavior Therapy (DBT), Functional Analytic Psychotherapy (FAP), and Mindfulness Based Cognitive Therapy (IBCT). This study reports a survey of clinicians and non-clinicians who self-identify with second or third wave approaches, and a group of undergraduate psychology students intended to represent a layperson or folk psychological approach. Their preferences, in the context of 10 clinical vignettes, among 5 different therapeutic responses or interventions that included "ACT-like," "cognitive," and commonsense or "neutral" options were measured. Third wave-oriented respondents exhibited more consistency than others in their preference for interventions that match their self-identified theoretical orientation, however the author suggests that construction of the vignettes may have influenced this result.

Otimização acústica e análise numérica do escoamento ao redor de um conjunto cilindro-placa separadora.

RESUMO Simulações de aeroacústica computacional demandam uma quantidade considerável de tempo, o que torna complicada a realização de estudos paramétricos. O presente trabalho propõe uma metodologia viável para otimização aeroacústica. Através da análise numérica utilizando dinâmica dos fluidos computacional, foi estudada a aplicação de uma placa separadora desacoplada como método de controle passivo da esteira turbulenta de um cilindro e avaliou-se a irradiação de ruído causado pela interação do escoamento com ambos os corpos, empregando ferramentas de aeroacústica computacional baseadas no método de Ffowcs-Williams e Hawkings. Algumas abordagens distintas de metodologias de otimização de projeto foram aplicadas neste problema, com o objetivo de chegar a uma configuração otimizada que permita a redução do nível sonoro ao longe. Assim, utilizando uma ferramenta de otimização multidisciplinar, pode-se avaliar a capacidade de modelos heurísticos e a grande vantagem do emprego de algoritmos baseados em método de superfície de resposta quando aplicados em um problema não linear, pois requerem a avaliação de um menor número de alternativas para se obter um ponto ótimo. Além disso, foi possível identificar e agrupar os resultados em 5 clusters baseados em seus parâmetros geométricos, nível de pressão sonora global e o valor quadrático médio do coeficiente de arrasto, confirmando a eficiência da aplicação de placas separadoras longas desacopladas posicionadas próximas ao cilindro na estabilização da esteira turbulenta, enquanto que o posicionamento de placas acima de um espaçamento crítico aumentou o nível de pressão acústica irradiado devido à formação de vórtices no espaço entre o cilindro e a placa separadora.

Nasabh:nāme:hazarat:nabhī:mahamadh:mûstafā : manuscript, 1900?

Second and third chapters of the anonymous Hindustani work entitled "Nasabh:nāme:hazarat:nabhī:mahamadh:mûstafā" and various accounts.

Various Shiite devotional texts to be recited in religious assemblies (majālis) commemorating the events leading to the martyrdom of the Shii Imam Ḥusain b. ʻAlī (d. 860)] : manuscript, 1900?

Various Shiite devotional texts to be recited in religious assemblies (majālis) commemorating the events leading to the martyrdom of the Shii Imam Ḥusain b. ʻAlī (d. 680). Each majlis (there is twenty) begins with a synoptic heading. The beginning of 17th majlis suggests that some of these texts are based on Mahdī b. Abī Zarr al-Kāshānī an-Niraqī's "Muḥarriq al-qulub (a Shiite history of the martyrs of the Prophet's family).

Satgûr:jo:salûko : manuscript, 1890?

Local paper.