Protein-tyrosine kinase and protein-serine/threonine kinase expression in human gastric cancer cell lines

Protein kinases play key roles in cellular functions. They are involved in many cellular functions including; signal transduction, cell cycle regulation, cell division, and cell differentiation. Alterations of protein kinase by gene amplification, mutation or viral factors often induce tumor formation and tumor progression toward malignancy. The identification and cloning of kinase genes can provide a better understanding of the mechanisms of tumorigenesis as well as diagnostic tools for tumor staging. In this study, we have used degenerated polymerase-chain-reaction primers according to the consensus catalytic domain motifs to amplify protein kinase genes (protein-tyrosine kinase, PTK, and protein-serine/threonine kinase, PSK) from human stomach cancer cells. Following amplification, the protein kinase molecules expressed in the gastric cancer cells were cloned into plasmid vectors for cloning and sequencing. Sequence analysis of polymerase-chain-reaction products resulted in the identification of 25 protein kinases, including two novel ones. Expression of several relevant PTK/PSK genes in gastric cancer cells and tissues was further substantiated by RT-PCR using gene-specific primers. The identification of protein kinases expressed or activated in the gastric cancer cells provide the framework to understand the oncogenic process of stomach cancer.     

Detection of siderophores in growing cultures ofPseudomonas spp.

Summary The siderophores produced byPseudomonas fluorescens andP. chlororaphis were detected from the culture supernatants in MM9 and modified King's medium by the universal CAS assay at wavelengths 620–690 nm. The CAS assay was applied to detectPseudomonas siderophores directly in situ, during their production phase, in modified King's medium. Optimum results were detected with a final CAS concentration of 0.025 mM and an iron concentration of 1.25 M. The problems of the method are discussed with respect to the absorbance spectrum, the toxicity of the HDTMA detergent, the influence of the iron concentration and the complexity of media for siderophore production.     

MEMOR: A database of archeological human remains collections from Flanders,Belgium

The aim of this article is to describe a newly created open access database of archeological human remains collections from Flanders, Belgium. The MEMOR database ( www.memor.be ) was created to provide an overview of the current practices of loans, reburial, and the research potential of human skeletons from archeological sites currently stored in Flanders. In addition, the project aimed to provide a legal and ethical framework for the handling of human remains and was created around stakeholder involvement from anthropologists, geneticists, contract archeologists, the local, regional and national government agencies, local and national government, universities, and representatives of the major religions. The project has resulted in the creation of a rich database with many collections available for study. The database was created using the open-source Arches data management platform that is freely available for organizations worldwide to configure in accordance with their individual needs and without restrictions on its use. Each collection is linked to information about the excavation and the site the remains originate from, its size and time period. In addition, a research potential tab reveals whether any analyses were performed, and whether excavation notes are available with the assemblage. The database currently contains 742 collections, ranging in size from 1 to over 1000 individuals. New collections will continue to be added when new assemblages are excavated and studied. The database can also be expanded to include human remains collections from other regions and other material categories, such as archaeozoological collections.     

A new synthesis method for complex electric field patterning using a multichannel dense array system with applications in low-intensity noninvasive neuromodulation

Multichannel coil array systems offer precise spatiotemporal electronic steering and patterning of electric and magnetic fields without the physical movement of coils or magnets. This capability could potentially benefit a wide range of biomagnetic applications such as low-intensity noninvasive neuromodulation or magnetic drug delivery. In this regard, the objective of this work is to develop a unique synthesis method, that enabled by a multichannel dense array system, generates complex current pattern distributions not previously reported in the literature. Simulations and experimental results verify that highly curved or irregular (e.g., zig–zag) patterns at singular and multiple sites can be efficiently formed using this method. The synthesis method is composed of three primary components; a pixel cell (basic unit of pattern formation), a template array (“virtual array”: code that disseminates the coil current weights to the “physical” dense array), and a hexagonal coordinate system. Low-intensity or low-field magnetic stimulation is identified as a potential application that could benefit from this work in the future and as such is used as an example to frame the research.     

Potassium channel blockers inhibit adoptive transfer of experimental allergic encephalomyelitis by myelin-basic-protein-stimulated rat T lymphocytes

Agents which block T cell K⁺ currents can prohibit both proliferative and effector cell functions in T cells activated by mitogens or phorbol esters. This study examined the effects of some of these blocking agents on the immune responsiveness of guinea pig myelin basic protein (GPMBP)-reactive Lewis rat T lymphocytes, which are capable of mediating the adoptive transfer of experimental allergic encephalomyelitis (EAE), an accepted animal model for multiple sclerosis. Both the proliferative functions (DNA synthesis and cell blastogenesis) and the EAE transfer activities of GPMBP-reactive lymphocytes were examined following GPMBP-induced activation in the presence of agents shown to block the outwardly rectifying K⁺ current in these cells. At concentrations which completely inhibited DNA synthesis, as measured by [³H]thymidine incorporation, and cell blastogenesis, tetraethylammonium (TEA), 4-aminopyridine (4-AP) and methoxyverapamil (D600) completely blocked the subsequent adoptive transfer of EAE into naive syngeneic Lewis rats. The concentrations at which these blockers produced a 50% reduction in DNA synthesis were estimated to be 16, 1.6 and 32 µM for TEA, 4-AP and D-600, respectively, which were roughly equivalent to the EC₅₀ to block the K⁺ current. Apamine, a potent Ca²⁺-activated K⁺ channel blocker, at a concentration several orders of magnitude higher than is necessary to block Ca²⁺-activated K⁺ channels, reduced the maximal K⁺ conductance in GPMBP-reactive T cell K⁺ channels by about 20%, but did not alter either [³H]thymidine incorporation or the adoptive transfer of EAE. These results indicate that delayed rectifier K⁺ channel blockers may prevent the activation of GPMBP-reactive T cells, thus prohibiting encephalitogenic effector cell functions.     

Acetyl-l-carnitine attenuates neuronal damage in gerbils with transient forebrain ischeia only when givenBefore the insult

The underlying mechanisms leading to neuronal damage in cerebral ischemia are multifactoral. In this study, we evaluated the neuroprotective effects of acetyl-l-carnitine, a medication that may enhance metabolic recovery after cerebral ischemia. The 5-minute transient forebrain ischemia model in gerbils was used. Acetyl-l-carnitine was given 30 minutes before the insult in one set of animals and 30 minutes after the insult in a second set of animals with histological evaluation at 7 days (Group A) and 28 days (Group B). Damage assessment was done using a 4-point damage score and Mann-Whitney U test was used for statistical analysis. Compared to the controls, there was significant protection in the cerebral cortex, hippocampus and the striatum in animals treated with the medicationbefore the insult in Group A and Group B Post-ischemic therapy showed little evidence of neuronal protection in either group. Behavioral tests in the Group B animals showed no significant differences between the treated or the saline controls. Our study shows, that pre-ischemic treatment with acetyl-l-carnitine results in neuronal protection. This may have clinical significance in situations (such as bypass surgery) where treatment could be initiatedprior to the insult.     

Preface

"Although few people die of pain,many people die in pain and even more live in pain"——IASP On October11,2004,International Association for the Study of Pain(IASP)     

Evaluating conditions for producing spray-dried formulations of Anagrapha falcifera nucleopolyhedroviruses (AfMNPV)

We investigated how altering parameters during the production of spray-dried lignin formulations affected the insecticidal activity of a baculovirus (AfMNPV), which was originally isolated from celery looper, Anagrapha falcifera (Kirby). Exposure to high temperature, varied pH of the dryer feedstock, substitution of formulation ingredients, and different commercial production lots of virus were evaluated for their effect on the insecticidal activity of AfMNPV. Insecticidal activities of treatments were determined by droplet-feeding assays using neonate Trichoplusia ni (Hübner) for dosage response or single-dosage comparisons. Unformulated virus exposed to 68^oC for 2 h showed no loss of insecticidal activity, whereas exposure to 90^oC for 30 min caused >20% loss of activity. Thus, residence at higher temperatures in drying systems could adversely affect virus activity. Spray-dried formulations made with Indulin AT lignin (pH 8.5, 9.5, and 10.5) lost insecticidal activity with increasing alkalinity of the dryer feedstock. In contrast to Indulin AT, the same formulations made with PC-1307 lignin did not lose insecticidal activity with increased alkalinity. Adding corn flour to spray-dried Indulin AT-based formulations improved insecticidal activity of the virus. These experiments demonstrated the importance of carefully selecting feed-stock ingredients and processing conditions when spray drying AfMNPV.