Self-renewal of primitive human hematopoietic cells (long-term-culture-initiating cells) in vitro and their expansion in defined medium.

A major goal of experimental and clinical hematology is the identification of mechanisms and conditions that support the expansion of transplantable hematopoietic stem cells. In normal marrow, such cells appear to be identical to (or represent a subset of) a population referred to as long-term-culture-initiating cells (LTC-ICs) so-named because of their ability to produce colony-forming cell (CFC) progeny for > or = 5 weeks when cocultured with stromal fibroblasts. Some expansion of LTC-ICs in vitro has recently been described, but identification of the factors required and whether LTC-IC self-renewal divisions are involved have remained unresolved issues. To address these issues, we examined the maintenance and/or generation of LTC-ICs from single CD34+ CD38- cells cultured for variable periods under different culture conditions. Analysis of the progeny obtained from cultures containing a feeder layer of murine fibroblasts engineered to produce steel factor, interleukin (IL)-3, and granulocyte colony-stimulating factor showed that approximately 20% of the input LTC-ICs (representing approximately 2% of the original CD34+ CD38- cells) executed self-renewal divisions within a 6-week period. Incubation of the same CD34+ CD38- starting populations as single cells in a defined (serum free) liquid medium supplemented with Flt-3 ligand, steel factor, IL-3, IL-6, granulocyte colony-stimulating factor, and nerve growth factor resulted in the proliferation of initial cells to produce clones of from 4 to 1000 cells within 10 days, approximately 40% of which included > or = 1 LTC-IC. In contrast, in similar cultures containing methylcellulose, input LTC-ICs appeared to persist but not divide. Overall the LTC-IC expansion in the liquid cultures was 30-fold in the first 10 days and 50-fold by the end of another 1-3 weeks. Documentation of human LTC-IC self-renewal in vitro and identification of defined conditions that permit their extensive and rapid amplification should facilitate analysis of the molecular mechanisms underlying these processes and their exploitation for a variety of therapeutic applications.

The site of action of neuronal acidic fibroblast growth factor is the organ of Corti of the rat cochlea.

Here we show that the mature cochlear neurons are a rich source of acidic fibroblast growth factor (aFGF), which is expressed in the neuronal circuitry consisting of afferent and efferent innervation. The site of action of neuronal aFGF is likely to reside in the organ of Corti, where one of the four known FGF receptor (FGFR) tyrosine kinases--namely, FGFR-3 mRNA--is expressed. Following acoustic overstimulation, known to cause damage to the organ of Corti, a rapid up-regulation of FGFR-3 is evident in this sensory epithelium, at both mRNA and protein levels. The present results provide in vivo evidence for aFGF being a sensory neuron-derived, anterogradely transported factor that may exert trophic effects on a peripheral target tissue. In this sensory system, aFGF, rather than being a neurotrophic factor, seems to promote maintenance of the integrity of the organ of Corti. In addition, aFGF, released from the traumatized nerve endings, may be one of the first signals initiating protective recovery and repair processes following damaging auditory stimuli.

Sphingosine: a mediator of acute renal tubular injury and subsequent cytoresistance.

The goal of this study was to determine whether sphingosine and ceramide, second messengers derived from sphingolipid breakdown, alter kidney proximal tubular cell viability and their adaptive responses to further damage. Adult human kidney proximal tubular (HK-2) cells were cultured for 0-20 hr in the presence or absence of sphingosine, sphingosine metabolites (sphingosine 1-phosphate, dimethylsphingosine), or C2, C8, or C16 ceramide. Acute cell injury was assessed by vital dye exclusion and tetrazolium dye transport. Their subsequent impact on superimposed ATP depletion/Ca2+ ionophore-induced damage was also assessed. Sphingosine (> or = 10 microM), sphingosine 1-phosphate, dimethylsphingosine, and selected ceramides (C2 and C8, but not C16) each induced rapid, dose-dependent cytotoxicity. This occurred in the absence of DNA laddering or morphologic changes of apoptosis, suggesting a necrotic form of cell death. Prolonged exposure (20 hr) to subtoxic sphingosine doses (< or = 7.5 microM) induced substantial cytoresistance to superimposed ATP depletion/Ca2+ ionophore-mediated damage. Conversely, neither short-term sphingosine treatment (< or = 8.5 hr) nor 20-hr exposures to any of the above sphingosine/ceramide derivatives/metabolites or various free fatty acids reproduced this effect. Sphingosine-induced cytoresistance was dissociated from the extent of cytosolic Ca2+ loading (indo-1 fluorescence), indicating a direct increase in cell resistance to attack. We conclude that sphingosine can exert dual effects on proximal renal tubular viability: in high concentrations it induces cell necrosis, whereas in low doses it initiates a cytoresistant state. These results could be reproduced in human foreskin fibroblasts, suggesting broad-based relevance to the area of acute cell injury and repair.

The causes and prevention of cancer.

Epidemiological evidence indicates that avoidance of smoking, increased consumption of fruits and vegetables, and control of infections will have a major effect on reducing rates of cancer. Other factors include avoidance of intense sun exposure, increases in physical activity, and reduction of alcohol consumption and possibly red meat. A substantial reduction in breast cancer is likely to require modification of sex hormone levels, and development of practical methods for doing so is a high research priority. Resolution of the potential protective roles of specific antioxidants and other constituents of fruits and vegetables deserves major attention. Mechanistic studies of carcinogenesis indicate an important role of endogenous oxidative damage to DNA that is balanced by elaborate defense and repair processes. Also key is the rate of cell division, which is influenced by hormones, growth, cytotoxicity, and inflammation, as this determines the probability of converting DNA lesions to mutations. These mechanisms may underlie many epidemiologic observations.

Regulatory region in choline acetyltransferase gene directs developmental and tissue-specific expression in transgenic mice.

Acetylcholine, one of the main neurotransmitters in the nervous system, is synthesized by the enzyme choline acetyltransferase (ChAT; acetyl-CoA:choline O-acetyltransferase, EC 2.3.1.6). The molecular mechanisms controlling the establishment, maintenance, and plasticity of the cholinergic phenotype in vivo are largely unknown. A previous report showed that a 3800-bp, but not a 1450-bp, 5' flanking segment from the rat ChAT gene promoter directed cell type-specific expression of a reporter gene in cholinergic cells in vitro. Now we have characterized a distal regulatory region of the ChAT gene that confers cholinergic specificity on a heterologous downstream promoter in a cholinergic cell line and in transgenic mice. A 2342-bp segment from the 5' flanking region of the ChAT gene behaved as an enhancer in cholinergic cells but as a repressor in noncholinergic cells in an orientation-independent manner. Combined with a heterologous basal promoter, this fragment targeted transgene expression to several cholinergic regions of the central nervous system of transgenic mice, including basal forebrain, cortex, pons, and spinal cord. In eight independent transgenic lines, the pattern of transgene expression paralleled qualitatively and quantitatively that displayed by endogenous ChAT mRNA in various regions of the rat central nervous system. In the lumbar enlargement of the spinal cord, 85-90% of the transgene expression was targeted to the ventral part of the cord, where cholinergic alpha-motor neurons are located. Transgene expression in the spinal cord was developmentally regulated and responded to nerve injury in a similar way as the endogenous ChAT gene, indicating that the 2342-bp regulatory sequence contains elements controlling the plasticity of the cholinergic phenotype in developing and injured neurons.

Fish diversity and traditional fishing methods in semiarid irrigated system in the Bajo Vinalopó region, Southeastern Spain

The aim of this paper is to obtain a comprehensive picture of the importance of the fish fauna in the wetlands and coastal lagoons of the Bajo Vinalopó region (Alicante, South eastern Spain) and the ecological, historical and cultural value of the traditional knowledge on fishing maintained by local people. We have compiled data from different information sources: (1) the revision of historical archives, (2) personal interviews and collaborative research, and (3) ecological and water quality data. The results show that the area sustains important fish fauna diversity (17 species) and that local people have inherited a considerable traditional knowledge on fishing methods (11 modalities), on the maintenance and sound management of the water system and on the ecology and behaviour of fish. We conclude that a comprehensive consideration of all these ecological, historical and socio-cultural aspects related to fish and fishing shows clearly the value of this ecological and cultural heritage and provides a necessary base for a sustainable management of the area.

Profiles of emotional intelligence and learning strategies in a sample of Chilean students

In the last few years, one of the lines of research of great interest in the field of emotional intelligence (EI) has been the analysis of the role of emotions in the educational context and, in particular, their influence on learning strategies. The aims of this study are to identify the existence of different EI profiles and to determine possible statistically significant differences in learning strategies between the obtained profiles. The study involved 1253 Chilean school students from 14 to 18 years (M = 15.10, SD = 1.30), who completed the Trait Meta-Mood Scale-24 (TMMS-24) and the Inventory of Learning and Study Strategies—High School version (LASSI-HS). Cluster analysis identified four EI profiles: a group of adolescents with a high EI profile, a group with predominance of low emotional attention and high repair skills, a group with high scores on attention and low scores on clarity and repair, and a final group of adolescents with low EI. Also, students in groups with high overall scores in EI and low attention and high repair emotional obtained higher scores on the different learning strategies; however, the effect size analysis showed that these differences had no empirical relevance.

Floating debris control : a literature review /

"June 1987."

Elements of floating-debris control systems /

"September 1988."

Design criteria for lateral dikes in estuaries /

At head of title: "Repair, Evaluation, Maintenance, and Rehabilitation Research Program."

STREMR : numerical model for depth-averaged incompressible flow /

"September 1993."

HIVEL2D : a two-dimensional flow model for high-velocity channels /

At head of title: "Repair, Evaluation, Maintenance, and Rehabilitation Research Program."

Breathing apparatus, rescue and protective.

Mode of access: Internet.

Digital Classics Outside the Echo-Chamber : Teaching, Knowledge Exchange and Public Engagement /

The international perspectives on these issues are especially valuable in an increasingly connected, but still institutionally and administratively diverse world. The research addressed in several chapters in this volume includes issues around technical standards bodies like EpiDoc and the TEI, engaging with ways these standards are implemented, documented, taught, used in the process of transcribing and annotating texts, and used to generate publications and as the basis for advanced textual or corpus research. Other chapters focus on various aspects of philological research and content creation, including collaborative or community driven efforts, and the issues surrounding editorial oversight, curation, maintenance and sustainability of these resources. Research into the ancient languages and linguistics, in particular Greek, and the language teaching that is a staple of our discipline, are also discussed in several chapters, in particular for ways in which advanced research methods can lead into language technologies and vice versa and ways in which the skills around teaching can be used for public engagement, and vice versa. A common thread through much of the volume is the importance of open access publication or open source development and distribution of texts, materials, tools and standards, both because of the public good provided by such models (circulating materials often already paid for out of the public purse), and the ability to reach non-standard audiences, those who cannot access rich university libraries or afford expensive print volumes. Linked Open Data is another technology that results in wide and free distribution of structured information both within and outside academic circles, and several chapters present academic work that includes ontologies and RDF, either as a direct research output or as essential part of the communication and knowledge representation. Several chapters focus not on the literary and philological side of classics, but on the study of cultural heritage, archaeology, and the material supports on which original textual and artistic material are engraved or otherwise inscribed, addressing both the capture and analysis of artefacts in both 2D and 3D, the representation of data through archaeological standards, and the importance of sharing information and expertise between the several domains both within and without academia that study, record and conserve ancient objects. Almost without exception, the authors reflect on the issues of interdisciplinarity and collaboration, the relationship between their research practice and teaching and/or communication with a wider public, and the importance of the role of the academic researcher in contemporary society and in the context of cutting edge technologies. How research is communicated in a world of instant- access blogging and 140-character micromessaging, and how our expectations of the media affect not only how we publish but how we conduct our research, are questions about which all scholars need to be aware and self-critical.

Safety Maintenance & Production

Title Varies: V.1-25,No.3, Insurance Engineering; V.25,No.4-V.100,No.3, Safety Engineering; V.100,No.4-V.112, Safety Maintenance and Production; V.113-138,No.3, Safety Maintenance; V.138,No.4-, Environmental Control Management