Differential acetylcholine and GABA release from cultured chick retina cells

In the present work we investigated the mechanisms controlling the release of acetylcholine (ACh) and of gamma-aminobutyric acid (GABA) from cultures of amacrine-like neurons, containing a subpopulation of cells which are simultaneously GABAergic and cholinergic. We found that 81.2 +/- 2.8% of the cells present in the culture were stained immunocytochemically with an antibody against choline acetyltransferase, and 38.5 +/- 4.8% of the cells were stained with an antibody against GABA. Most of the cells containing GABA (87.0 +/- 2.9%) were cholinergic. The release of acetylcholine and GABA was mostly Ca2+-dependent, although a significant release of [3H]GABA occurred by reversal of its transporter. Potassium evoked the Ca2+-dependent release of [3H]GABA and [3H]acetylcholine, with EC50 of 31.0 +/- 1.0 mm and 21.6 +/- 1.1 mm, respectively. The Ca2+-dependent release of [3H]acetylcholine was significantly inhibited by 1 micrometer tetrodotoxin and by low (30 nm) omega-conotoxin GVIA (omega-CgTx GVIA) concentrations, or by high (300 nm) nitrendipine (Nit) concentrations. On the contrary, the release of [14C]GABA was reduced by 30 nm nitrendipine, or by 500 nm omega-CgTx GVIA, but not by this toxin at 30 nm. The release of either transmitters was unaffected by 200 nm omega-Agatoxin IVA (omega-Aga IVA), a toxin that blocks P/Q-type voltage-sensitive Ca2+ channels (VSCC). The results show that Ca2+-influx through omega-CgTx GVIA-sensitive N-type VSCC and through Nit-sensitive L-type VSCC induce the release of ACh and GABA. However, the significant differences observed regarding the Ca2+ channels involved in the release of each neurotransmitter suggest that in amacrine-like neurons containing simultaneously GABA and acetylcholine the two neurotransmitters may be released in distinct regions of the cells, endowed with different populations of VSCC.     

Changes in bacterial species composition in enrichment cultures with various dilutions of inoculum as monitored by denaturing gradient gel electrophoresis

Denaturing gradient gel electrophoresis revealed changes in the bacterial species obtained from enrichment cultures with different inoculum dilutions. This inoculum dilution enrichment approach may facilitate the detection and isolation of a greater number of bacterial species than traditional enrichment techniques.     

The role of oncostatin M in animal and human connective tissue collagen turnover and its localization within the rheumatoid joint

OBJECTIVE: To study the interaction of interleukin-1alpha (IL-1alpha) and oncostatin M (OSM) in promoting cartilage collagen destruction. METHODS: Bovine, porcine, and human cartilage and human chondrocytes were studied in culture. The levels of collagenase (matrix metalloproteinase 1 [MMP-1]) and tissue inhibitor of metalloproteinases 1 (TIMP-1) were measured by bioassay and enzyme-linked immunosorbent assay (ELISA). The levels of OSM in rheumatoid synovial fluid were measured by ELISA. RESULTS: When combined with OSM, IL-1alpha, IL-1beta, and tumor necrosis factor alpha released proteoglycan and collagen from cartilage. OSM was the only member of the IL-6 family to have this effect. Human tendon also responded to IL-1alpha and OSM. OSM increased the production of MMP-1 and TIMP-1 but when combined with IL-1alpha, synergistically promoted MMP-1 production in human chondrocytes and synovial fibroblasts. High levels of OSM were found in human rheumatoid synovial fluids, and confocal microscopy showed that OSM was produced by macrophages in rheumatoid synovial tissue. CONCLUSION: These results highlight an important new mechanism by which there is irreversible loss of collagen from cartilage.     

Mesenteric venous thrombosis after appendicectomy in a child: clinical case and review of the literature

Mesenteric venous thrombosis (MVT) after appendicectomy has not been reported in the pediatric literature. This complication is associated with significant morbidity and mortality rates in adults. The report of this case attempts to analyze the features found in an appendicectomized child documented by radiological examination and surgery before and after starting heparin therapy. Anticoagulation treatment should be started as soon as possible and continued for a long time to decrease the risk of relapse.     

Production of human normal adult and fetal hemoglobins in Escherichia coli

A hemoglobin expression system in Escherichia coli is described. In order to produce authentic human hemoglobin, we need to co-express both methionine aminopeptidase and globin genes under the control of a strong promoter. We have constructed three plasmids, pHE2, pHE4 and pHE7, for the expression of human normal adult hemoglobin and a plasmid, pHE9, for the expression of human fetal hemoglobin, in high yields. The globin genes can be derived from either synthetic genes or human globin cDNAs. The extra amino-terminal methionine residues of the expressed globins can be removed by the co-expressed methionine aminopeptidase. The heme is inserted correctly into the expressed alpha- globin from our expression plasmids. A fraction (approximately 25%) of the heme is not inserted correctly into the expressed beta- or gamma- globin. However, the incorrectly inserted hemes can be converted into the correct conformation by carrying out a simple oxidation-reduction process on the purified hemoglobin molecule. We have investigated the functional properties of the expressed hemoglobins by measuring their oxygen-binding properties and their structural features by obtaining their 1H-NMR spectra. Our results show that authentic human normal adult and fetal hemoglobins can be produced from our expression plasmids in E. coli and in high yields. Our expression system allows us to design and to produce any recombinant hemoglobins needed for our research on the structure-function relationship in hemoglobin.      

A 931 + 2T-->C transition in one COL1A2 allele causes exon 16 skipping in PRO alpha 2(I) mRNA and produces moderately severe OI

The breast clinic in the university Hospital Kuala Lumpur was started in August 1993 to cater for the increasing number of women seeking treatment for breast complaints. A four-month survey carried out from January to April 1994 found that Chinese women are more "breast conscious" than Malay and Indian women. The majority of patients were in the reproductive age group. Benign breast diseases form the majority of cases of breast diseases seen. The pick-up rate for new cases of breast cancer was 3.1%. Fine needle aspiration biopsy had a 100% sensitivity during the four-month study, and was a quick method of obtaining a diagnosis.     

Future perspectives on sustainable tribology

This paper highlights the future perspectives of sustainable tribology by examining the economic, environmental and social impact of three tribological case studies. One case study examines the sustainability and durability of micro-CHP systems looking the tribological phenomena generated within a scroll expander system. The scroll is the main part of a specific micro-CHP system and experiences wear and cavitation damage. The tribological optimization of the scroll expander improves the sustainability of the micro-CHP unit while it has a serious economic and environmental impact to the consumers and to the society in general. Another case study is focused on friction and wear performance of lifeboat launch slipways. The causes of high friction and wear during the RNLI's lifeboat launches along an inclined slipway are investigated with a view to reducing the environmental impact due to slipway panel wear and lubricant release into the marine environment. The project encompasses the sustainable design of slipway panels using design modifications based on tribological investigations to double their lifespan, while environmental and economic impact was significantly reduced by the use of biodegradable greases and water as lubricants. The final case study involves an investigation of recycled plastic materials to replace polyurethane used on skateboard wheels, scooters and similar applications. Polyurethane (PU) is difficult to recycle. With the dwindling resources and environmental problems facing the world today, recycling for both waste reduction and resource preservation has become an increasingly important aspect of sustainability. The tribological results showed that recycled polycarbonate plastic can effectively act as a substitute to polyurethane wheels. Moreover, sustainability considerations showing the environmental benefits of the use of recycled plastics over PU include reducing the CO₂ footprint by 50% and the energy consumed by 60%, among other benefits. These case studies emphasise the importance of sustainable tribology in our epoch showing that increased sustainability performance can be achieved through tribology to a significant extent in many cases, providing stability to our world and more viable long term growth to our societies.     

An Algorithm for Projecting Points onto a Patched CAD Model

We are interested in building structured overlap-ping grids for geometries defined by Computer-Aided-Design (CAD) packages. Geometric information defining the boundary surfaces of a computation domain is often provided in the form of a collection of possibly hundreds of trimmed patches. The first step in building an overlapping volume grid on such a geometry is to build overlapping surface grids. A surface grid is typically built using hyperbolic grid generation; starting from a curve on the surface, a grid is grown by marching over the surface. A given hyperbolic grid will typically cover many of the underlying CAD surface patches. The fundamental operation needed for building surface grids is that of projecting a point in space onto the closest point on the CAD surface. We describe a fast and robust algorithm for performing this projection which makes use of a fairly coarse global triangulation of the CAD geometry. Before the global triangulation is constructed the connectivity of the model is determined by an edge-matching algorithm which corrects for gaps and overlaps between neighbouring patches. ID="A1" Correspondence and offprint requests to: Dr. W. D. Henshaw, Center for Applied Scientific Computing, L-661, Lawrence Livermore National Laboratory, Livermore, CA 94551, USA. E-mail: henshaw@llnl.gov     

Megavoltage imaging with low Z targets: implementation and characterization of an investigational system

The poor quality of stereotactic radiotherapy portal images is a limiting factor in precise image registration. To alleviate this problem, a low atomic number (Z) target was implemented on our Siemens MXE linear accelerator. This investigational system was used to assess the performance of various target materials by filming an aluminum contrast object. Beryllium, carbon and conventional target materials were studied. The bremsstrahlung spectra of these materials were simulated using Monte Carlo techniques. These spectra were used to calculate the dependence of narrow beam contrast on phantom thickness for verification of the data measured from film. A Monte Carlo simulation of the beryllium spectrum in a wide beam geometry was used to evaluate the effect of phantom-to-film distance on contrast. Although the same degree of contrast improvement with distance was not realized in practice, the improvement in image quality rivaled that achieved using a scatter reduction grid. A comparison of conventional localization images of the head and neck of an anthropomorphic phantom with images produced with a beryllium or carbon target and a mammography film and screen system supports earlier suggestions that the technique is clinically useful.