Techniques for live capture of deepwater fishes with special emphasis on the design and application of a low-cost hyperbaric chamber

A cost effective, simple, portable hyperbaric chamber was constructed from polyvinyl chloride to aid in the collection of adult rockfishes Sebastes sp. to hold as broodstock. This system was designed to recompress fishes quickly once brought to the surface on hook and line, and to allow for decompression over a period of days. The hyperbaric chamber is capable of continuous stable operation at <1 033 515 N m⁻² and can accommodate fishes up to 91·4 cm in length and 26·8 cm in diameter. Pressure in the chamber is maintained by a Goulds Booster pump that delivers continuous pressure and supplies sea water at a rate of 3·8 to 7·6 l min⁻¹ to as many as four chambers. The hyperbaric chamber operated very effectively and allowed successful decompression of 12 cowcod Sebastes levis captured at depths of 90·2 to 146·3 m.     

An inexpensive system for exposing plants in the field to elevated concentrations of CO2

An inexpensive, potentially mobile field exposure system is described which may be easily constructed by a small workshop. It may be operated as an open-top with a frustrum or covered with a polycarbonate ‘lid’. The system is cost-effective for CO₂ exposure work because the small size allows provision of CO₂-enriched atmospheres over prolonged periods at relatively low cost. A preliminary assessment of the chambers has been made and concentrations can be maintained at ±6% for a target atmosphere of 680 cm³ m^?3 CO₂ under normal operating conditions. Other chamber environmental conditions are reported.     

Correction of measured Gamma-Knife output factors for angular dependence of diode detectors and PinPoint ionization chamber

Dosimetry for Gamma-Knife requires detectors with high spatial resolution and minimal angular dependence of response. Angular dependence and end effect time for p-type silicon detectors (PTW Diode P and Diode E) and PTW PinPoint ionization chamber were measured with Gamma-Knife beams. Weighted angular dependence correction factors were calculated for each detector. The Gamma-Knife output factors were corrected for angular dependence and end effect time. For Gamma-Knife beams angle range of 84°–54°. Diode P shows considerable angular dependence of 9% and 8% for the 18 mm and 14, 8, 4 mm collimator, respectively. For Diode E this dependence is about 4% for all collimators. PinPoint ionization chamber shows angular dependence of less than 3% for 18, 14 and 8 mm helmet and 10% for 4 mm collimator due to volumetric averaging effect in a small photon beam. Corrected output factors for 14 mm helmet are in very good agreement (within ±0.3%) with published data and values recommended by vendor (Elekta AB, Stockholm, Sweden). For the 8 mm collimator diodes are still in good agreement with recommended values (within ±0.6%), while PinPoint gives 3% less value. For the 4 mm helmet Diodes P and E show over-response of 2.8% and 1.8%, respectively. For PinPoint chamber output factor of 4 mm collimator is 25% lower than Elekta value which is generally not consequence of angular dependence, but of volumetric averaging effect and lack of lateral electronic equilibrium. Diodes P and E represent good choice for Gamma-Knife dosimetry.     

Over a Century of Neuron Culture: From the Hanging Drop to Microfluidic Devices

The brain is the most intricate, energetically active, and plastic organ in the body. These features extend to its cellular elements, the neurons and glia. Understanding neurons, or nerve cells, at the cellular and molecular levels is the cornerstone of modern neuroscience. The complexities of neuron structure and function require unusual methods of culture to determine how aberrations in or between cells give rise to brain dysfunction and disease. Here we review the methods that have emerged over the past century for culturing neurons in vitro, from the landmark finding by Harrison (1910) — that neurons can be cultured outside the body — to studies utilizing culture vessels, micro-islands, Campenot and brain slice chambers, and microfluidic technologies. We conclude with future prospects for neuronal culture and considerations for advancement. We anticipate that continued innovation in culture methods will enhance design capabilities for temporal control of media and reagents (chemotemporal control) within sub-cellular environments of three-dimensional fluidic spaces (microfluidic devices) and materials (e.g., hydrogels). They will enable new insights into the complexities of neuronal development and pathology.     

Restricted distribution of laminin α1 chain in normal adult mouse tissues

The distribution of laminin α1 chain in adult mouse tissue was determined by immunofluorescence using monoclonal antibody 200, reacting with the globular carboxyterminus E3 fragment of α1 chain. Strong reactivity was noted only in a few tissues. Reactivity was restricted to epithelial basement membranes. Expression was noted in several epithelial basement membranes of the urinary tract, and male and female reproductive organs. In addition, expression was seen in some parts of the nervous system. Expression was seen in pia mater which surrounds the brain, and in the extracellular matrices covering the vitreous chamber and the lens of the eye. Staining was seen in the adrenal gland cortex, with strongest staining in the zona glomerulosa. Staining was negative in all other studied epithelial basement membranes, such as the lung (trachea or lung epithelium), epidermis, and all parts of the gastrointestinal tract (liver, gut) except for weak staining in the ventricle and Brunner’s glands. No expression was seen in basement membranes of fat, Schwann, or endothelial cells in any studied parts of the body. Both small- and large-size vessel walls were negative both in endothelial basement membranes and blood vessel walls, with the exception of some larger brain blood vessels in locations where epithelial cells have invaginated. Neither smooth muscle, myocardium or striated muscle expressed α1 chain. We conclude that α1-containing heterotrimers including laminin-1 (α1β1γ1) have a very restricted tissue distribution.     

Water uptake by some chamber materials

Abstract. The characteristics of water uptake by several materials have been studied. Nitrile rubber, silicone rubber and perspex retained substantial quantities of water. Metals took up smaller, but still significant, amounts of water, and glass was the least water-retentive material used in the tests. The surface structure of the material, revealed by scanning electron microscopy, does not necessarily account for its water uptake characteristics. The results are discussed in relation to the design and construction of psychrometer and porometer chambers.     

Short-term responses of phenology, shoot growth and leaf traits of four alpine shrubs in a timberline ecotone to simulated global warming, Eastern Tibetan Plateau, China

The short-term effects of artificial warming on phenology, growth and leaf traits were investigated in four alpine shrubs using the open-top chamber (OTC) method in a timberline ecotone (3240 m a.s.l.) on the Eastern Tibetan Plateau. The OTC enhanced the mean air temperature by 2.9°C throughout the growing season. In contrast, only a slight difference (0.4°C) in the mean soil temperature was observed in the OTC compared with the control plots (CP). Spiraea mongolica , Potentilla fruticosa , Conicera hispida (deciduous shrubs) and Daphne retusa (evergreen shrub) showed earlier bud break, flowering and fruit coloring as well as longer flower longevity in the OTC than in the CP. All deciduous shrubs in the OTC had a longer leaf lifespan. Daphne retusa had higher leaf survival rates in the OTC. No significant differences in the total number of flowers and fruits were noticed for most species between the two treatments. Warming stimulated the shoot and leaf growth for most species. The specific leaf area tended to increase for many species in the OTC. However, the leaf nitrogen concentration tended to decrease in P. fruticosa and S. mongolica. The results obtained in the present study indicate that warming conditions can have strong impacts on alpine shrubs in a timberline ecotone.     

Perfusion culture system: Synovial fibroblasts modulate articular chondrocyte matrix synthesis in vitro

Objective

To investigate the interactions of chondrocyte metabolism by synovial cells and synovial supernatants in a new perfusion co-culture system.

Methods

Chondrocytes and synovial fibroblasts were obtained from knee joints of slaughtered adult cattle. For experimental studies chondrocytes and synovial fibroblasts were placed together into a perfusion chamber (co-culture) or were placed into two different perfusion culture containers, which were connected by a silicone tube (culturing of chondrocytes with synovial supernatants). A control setup was used without synovial cells. Chondrocyte proliferation was shown by measurement of DNA content. The proteoglycan synthesis was quantified using ³⁵SO₄²⁻-labelling and the dimethylmethylene blue assay. ³H-proline incorporation was used to estimate the protein biosynthesis. Type II collagen synthesis was measured by ELISA, furthermore extracellular matrix deposition was monitored immunohistochemically (collagen types I/II). Regarding to the role of reactive oxygen species LDH release before and after stimulation with hydrogen peroxide was measured.

Results

The proliferation of chondrocytes shows an increase in monoculture as well as in co-culture or in culture with synovial supernatants more than fivefold within 12 days. ³H-proline incorporation as a marker for chondrocytes biosynthetic activity decreases in co-culture system and in culture with synovial supernatants. A similar effect is seen measuring total proteoglycan content as well as the ³⁵SO₄²⁻ incorporation in chondrocytes. Co-culturing and culturing with synovial supernatants lead to a significant decrease of proteoglycan release and content. Quantification of collagen type II by ELISA shows significant lower amounts of native collagen type II in the extracellular matrix of co-cultured chondrocytes as well as in culture with synovial supernatants. The membrane damage of chondrocytes by hydrogen peroxide is reduced when chondrocytes are co-cultured with synovial fibroblasts.

Conclusion

The co-culture perfusion system is a new tool to investigate interactions of different cell types with less artificial interferences. Our results suggest that synovial supernatants and synovial fibroblasts modulate the biosynthetic activity and the matrix deposition of chondrocytes as well as the susceptibility to radical attack of reactive oxygen species.     

Effects of aquatic macrophytes on organic matter deposition,resuspension and phosphorus entrainment in a lowland river

1. Although macrophytes play a key role in the structure and functioning of lowland rivers, most of the basic plant, hydrodynamic and sediment‐water interactions have only been described qualitatively. We therefore studied quantitatively, the seasonal dynamics of matter deposition and mobilisation inside and outside (free path) a representative patch of arrowhead, Sagittaria sagittifolia, in the lowland River Spree, NE Germany, in August 2006. Our in situ study combined resuspension experiments, a hydrodynamically calibrated erosion chamber and concurrent measurements of the prevailing flow characteristics and bed load. 2. Increasing entrainment rates (E) of particles (E_SPM) and total P (E_TP), with increments of shear velocity (U_*) from 0.53 to 2.42 cm s^?1, were significantly higher inside the plant patch than outside. Indeed, E_SPM and E_TP at the lowest U_* were 8‐ and 12‐fold higher inside than outside the patch, reflecting the resuspension potential of the upper nutrient‐enriched layer and the extent of pulsed P inputs even at small increases in U_*. 3. Vertical distribution of velocity (u) revealed a flow pattern of a mixing layer inside the S. sagittifolia patch, and that of a boundary layer in the free path. The highest gradient of u in the mixing layer was located in the water column at about 0.5 m depth, whereas the highest gradient of u for the boundary layer was found near the riverbed. The maximum of U_* (1.65 cm s^?1) was only 4 mm above the sediment. 4. A plant mosaic provides a low‐energetic environment promoting extensive particle trapping and the accumulation of a fine‐grained, nutrient‐enriched sediment, and forming a large resuspension potential. Consequently, after plant decay and the concomitant increase of U_* this material is preferentially entrained at higher rates. Hence, the key role of submerged macrophytes in lowland rivers is more directly related to modifying the dynamic equilibria between vegetation trapping and resuspension, than to the retention of nutrients, particularly P, and the reduction of P loads downstream to other waters.