Netrin-3 Peptide (C-19) is a Chemorepellent and a Growth Inhibitor in \u3cem\u3eTetrahymena thermophila\u3c/em\u3e

The netrins are a family of signaling proteins expressed throughout the animal kingdom. Netrins play important roles in developmental processes such as axonal guidance and angiogenesis. Netrin-1, for example, can act as either a chemoattractant or a chemorepellent for axonal growth cones depending upon the concentration of the protein as well as the cell type. Netrin-1 acts as a growth factor in some cell types and is expressed by some tumor cells. Netrin-3 appears to share some signaling apparatus with netrin-1, but is less widely expressed, and its physiological roles are much less understood. Tetrahymena thermophila are free-living, eukaryotic, ciliated protozoa used as a model system for studying chemorepellents and chemoattractants because their swimming behavior is readily observable under a microscope. We have previously found that netrin-1 peptide acts as a chemorepellent in Tetrahymena thermophila at concentrations ranging from micromolar to nanomolar. However, netrin-1 peptide does not affect growth in Tetrahymena at these concentrations. In our current study, we have found that related peptides, netrin-3 peptide (H-19 and C-19; Santa Cruz Biotechnology), act as chemorepellents in Tetrahymena thermophila at concentrations at or below 1 μg/ml. The same concentration of netrin-3 peptide reduces growth of Tetrahymena cultures by approximately 75%. We are currently conducting further studies to determine the mechanism through which these peptides are signaling

A Netrin-3 Like Protein is Secreted from \u3ci\u3eTetrahymena thermophila\u3c/i\u3e

Netrin proteins are a family of laminin-related secreted proteins that provide signals for axonal growth and cell migration during vertebrate development. Netrin homologs are expressed throughout the animal kingdom; however, some animals do not express a homolog of any known netrin receptors. We have previously found that the ciliated protozoan, Tetrahymena thermophila, responds to netrin-1 peptide by showing avoidance behavior. In addition, Tetrahymena secrete a protein that is immunologically similar to netrin-1 as detected by ELISA. Since netrin-3, like netrin-1, is a guidance molecule for axons and overlaps signaling pathways with netrin-1 in vertebrates, we hypothesized that netrin-3 may also act as a chemorepellent in Tetrahymena. While behavioral assays did not confirm this hypothesis, growth assays indicate that netrin-3 peptide inhibits cell division in Tetrahymena. In addition, ELISA and Western blots indicate that a netrin-3 like protein of approximately 48 kDa is secreted from Tetrahymena. Immunolocalization of this protein within the cell shows that it appears in widely distributed throughout the cell, and co-localizes with the netrin-1 like protein. Using ER tracker™, we found that some of the netrin-3-like protein co-localizes with the endoplasmic reticulum, as might be expected for a secreted protein. Further experimentation is necessary to determine the mechanism by which netrin-3 peptide inhibits growth in Tetrahymena

Mapping Netrin Signaling in \u3cem\u3eTetrahymena thermophila\u3c/em\u3e

The netrin family of proteins, found throughout the animal kingdom, are well known for their roles in developmental signaling. Netrin-1, the best-studied member of this family, signals through four receptor types in vertebrates: the UNC-5 family, DCC, neogenin, and DSCAM. We have previously characterized a netrin-1-like protein in the ciliated protozoan, Tetrahymena thermophila. This protein is secreted from Tetrahymena, and functions as a chemorepellent. Since a netrin-like protein is produced by this organism, we hypothesized that some components of the vertebrate netrin signaling pathway might also be present in Tetrahymena. Through immunolocalization on the plasma membrane of the cell, we have found that Tetrahymena appear to have a UNC-5 like protein, as well as proteins that are immunologically similar to neogenin. A homolog of src-1, a tyrosine kinase involved in vertebrate netrin-1, is also present in Tetrahymena. Future experiments will allow us to make more comparisons between netrin signaling in Tetrahymena with netrin signaling in the animal kingdom, and will allow us to determine the suitability of Tetrahymena as a model system for this particular pathway

Home Vegetable Gardening in Kentucky

Before You Begin Every aspiring gardener should follow seven steps to have a successful gardening season: Plan your garden on paper before you begin. Select a good gardening site that is: a. in full sun for at least eight hours each day, b. relatively level, c. well‑drained, d. close to a water source, e. not shaded. Prepare the soil properly and add fertilizer and lime according to soil test recommendations. Plan only as large a garden as you can easily maintain. Beginning gardeners often overplant, and then they fail because they cannot keep up with the tasks required. Weeds and pests must be controlled, water applied when needed and harvesting done on time. Vegetables harvested at their peak are tasty, but when left on the plants too long, the flavor is simply not there. Grow vegetables that will produce the maximum amount of food in the space available. Plant during the correct season for the crop. Choose varieties recommended for your area. Harvest vegetables at their proper stage of maturity. Store them promptly and properly if you do not use them immediately

Comparison of patterns of use, beliefs, and attitudes related to waterpipe between beginning and established smokers

BACKGROUND: To compare patterns of use, beliefs, and attitudes related to waterpipe smoking between university students (beginning smokers) and café customers (established smokers) in Aleppo Syria, in order to explore the evolution of this smoking method. METHODS: Two cross-sectional surveys were conducted among representative samples of university students (total 587, 48.4% men, mean age 22 years), and waterpipe users among cafe' customers (total 268, 60% men, mean age 30 years) in Aleppo, Syria. We used interviewer-administered questionnaire inquiring about pattern of waterpipe smoking (initiation, frequency), situational characteristics of use (partner, place, sharing), beliefs related to waterpipe smoking (harmful/addictive properties of waterpipe), attitudes related to waterpipe smoking (confidence in quitting, will to quit, motivation for quitting, past year quit attempt), and cigarette smoking. RESULTS: Daily and regular patterns of smoking become more prevalent with increased duration of smoking, but intermittent smoking remains the predominant pattern of waterpipe use. Women seem to be drawn later to the habit, which seem to escape the usual taboo against women's cigarette smoking. Patterns and context of waterpipe use tend to change with progress of the practice affecting frequency, setting, and sharing of waterpipe. Unlike beginners, established waterpipe smokers seem more smoking-method oriented, more hooked on the habit, less willing to quit, and less likely to foresee challenges to quitting. CONCLUSION: Use patterns and attitudes related to waterpipe smoking evolve to accommodate the change in dependence and life circumstances of the smoker. Most of use features, beliefs, attitudes, as well as time-course seem unique to this smoking method requiring novel approach to intervention

Netrin-3 Peptide (C-19) is a Chemorepellent and a Growth Inhibitor in \u3cem\u3eTetrahymena thermophila\u3c/em\u3e

The netrins are a family of signaling proteins expressed throughout the animal kingdom. Netrins play important roles in developmental processes such as axonal guidance and angiogenesis. Netrin-1, for example, can act as either a chemoattractant or a chemorepellent for axonal growth cones depending upon the concentration of the protein as well as the cell type. Netrin-1 acts as a growth factor in some mammalian cell types and is also expressed by some tumor cells. Netrin-3 appears to share some signaling apparatus with netrin-1, but is less widely expressed, and its physiological roles are much less understood. Netrin-3 is also used as a biomarker for some cancers as well as traumatic kidney injury. Tetrahymena thermophila are free-living, eukaryotic, ciliated protozoas used as a model system for studying chemorepellents and chemoattractants because their swimming behavior is readily observable under a microscope. We have previously found that netrin-1 peptide acts as a chemorepellent in Tetrahymena thermophila at concentrations ranging from micromolar to nanomolar. However, netrin-1 peptide does not affect growth in Tetrahymena at these concentrations. In our current study, we have found that related peptides, netrin-3 peptide (H-19 and C-19; Santa Cruz Biotechnology), act as chemorepellents in Tetrahymena thermophila at concentrations at or below 1 μg/ml. The same concentration of netrin-3 peptide reduces growth of Tetrahymena cultures by approximately 75%. We are currently conducting further studies to determine the mechanism through which these peptides are signaling

Glucose regulation in non-insulin-dependent diabetes mellitus: Interaction between pancreatic islets and the liver

The degree of fasting hyperglycemia in patients with non-insulin-dependent diabetes mellitus is dependent on the rate of hepatic glucose production. The basal rate of hepatic glucose production is increased in patients with non-insulin-dependent diabetes mellitus, and there is a positive correlation between hepatic glucose production and fasting glucose levels. Diminished secretion of insulin, impaired hepatic sensitivity to insulin's effects, or a combination of these factors could contribute to the elevated hepatic glucose production in patients with non-insulin-dependent diabetes mellitus. The relationship between insulin secretion and hepatic glucose production is regulated by a closed feedback loop operating between glucose levels and pancreatic beta cells. Although fasting insulin levels are usually comparable between patients with non-insulin-dependent diabetes mellitus and normal subjects, insulin secretion is markedly impaired in non-insulin-dependent diabetes mellitus in relation to the degree of hyperglycemia present. In fact, the degree of fasting hyperglycemia in a given patient with non-insulin-dependent diabetes mellitus is closely related to the degree of impaired pancreatic beta-cell responsiveness to glucose. Such findings suggest that impaired insulin secretion leads to increased hepatic glucose production, which raises the plasma glucose level. The resulting hyperglycemia helps to maintain relatively normal basal insulin output. Chronic sulfonylurea drug therapy of patients with non-insulin-dependent diabetes mellitus enhances pancreatic islet sensitivity to glucose, leading to increased insulin secretion, suppression of hepatic glucose production, and a decline in the steady-state fasting glucose level.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/25595/1/0000139.pd

Netrin-3 Avoidance and Mitotic Inhibition in \u3cem\u3eTetrahymena thermophila\u3c/em\u3e Involves Intracellular Calcium and Serine/Threonine Kinase Activity

Netrins are a family of signaling proteins ubiquitously expressed throughout the animal kingdom. While netrin-1 has been well characterized, other netrins, such as netrin-3, remain less well understood. In our current study, we characterize the behavior of two netrin-3 peptides, one derived from the N-terminal and one derived from the C-terminal of netrin-3. Both peptides cause avoidance behavior and mitotic inhibition in Tetrahymena thermophila at concentrations as low as 0.5 micrograms (μg) per milliliter. These effects can be reversed by addition of the calcium chelator, EGTA; the intracellular calcium chelator, BAPTA-AM, or the serine/threonine kinase inhibitor, apigenin. The broad spectrum tyrosine kinase inhibitor, genistein, has no effect on netrin-3 signaling, indicating that netrin-3 signaling in this organism uses a different pathway than the previously described netrin-1 pathway. Further studies will allow us to better describe the netrin-3 signaling pathway in this organism

Injuries in Aleppo, Syria; first population-based estimates and characterization of predominant types

BACKGROUND: Despite the growing burden of injuries worldwide, Syria and many other Arab countries still lack population-based estimates of different types of injuries. This study aims toprovide first population-based estimates of major injuries in Syria and characterize groups at increased risk. METHODS: An interviewer-administered population-based survey of adults 18–65 years residing in Aleppo, Syria was conducted in 2004. The study sample involved 2038 household representatives in Aleppo (45.2% men, mean age 35.3 ± 12.1, response rate 86%). We inquired about participants self-reported injuries in the past year that required medical attention as well as injuries among their household members. When reported, injuries were further assessed according to type, place, and outcome. RESULTS: Overall, there was 153 self-reported injuries in the past year (77.3 per 1000 adult respondents, 93.1 per 1000 in men and 64.4 per 1000 in women, p = 0.02). Other than gender, injuries differed by age (the older age group being least affected), and place of occurrence, as men were more likely to sustain traffic injuries and be injured outside the home. Injuries were reported among 236 household members (21.0 per 1000), and were slightly more frequent in children than adults (22.0 per 1000 for children, and 19.7 per 1000 for adults, p = 0.2). Traffic injuries, falls, and poisoning (food) were by far the most common types of injury experienced by participants as well as their household members. Falls and traffic injuries seem to have caused most morbidity for the injured, while burns, although not frequently reported, were associated with an unfavorable outcome in the majority of cases. CONCLUSION: This information provides baseline information about the burden of different injuries in Syria, and the sociodemographic factors related to them

Partial costs of global climate change adaptation for the supply of raw industrial and municipal water: a methodology and application

Despite growing recognition of the importance of climate change adaptation, few global estimates of the costs involved are available for the water supply sector. We present a methodology for estimating partial global and regional adaptation costs for raw industrial and domestic water supply, for a limited number of adaptation strategies, and apply the method using results of two climate models. In this paper, adaptation costs are defined as those for providing enough raw water to meet future industrial and municipal water demand, based on country-level demand projections to 2050. We first estimate costs for a baseline scenario excluding climate change, and then additional climate change adaptation costs. Increased demand is assumed to be met through a combination of increased reservoir yield and alternative backstop measures. Under such controversial measures, we project global adaptation costs of $12 bn p.a., with 83-90$73 bn p.a.), which supports the notion of mainstreaming climate change adaptation into broader policy aims. The method provides a tool for estimating broad costs at the global and regional scale; such information is of key importance in international negotiations. © 2010 IOP Publishing Ltd