Magnetic seeding to aid recovery of biological precipitates

A new approach to solid/liquid separation for biological precipitates is presented. The precipitate is seeded with small dense and/or magnetic particles to increase the density difference between precipitate and liquid or to make the precipitate amenable to magnetic separation. Experiments on seeding of ammonium sulphate precipitates of casein and separation of the seeded precipitate by gravity settling and batch centrifugation have shown that the approach holds promise. Seeded precipitates exhibit hindered settling under gravity with typical settling velocities of 0.6 cm/min for iron oxide seed and 2.4 cm/min for nickel seed. Calculations indicate that seeding of these precipitates causes a 23 fold increase in density difference between precipitate particle and supernatant liquid. The design of a magnetic seeding step for use in an enzyme isolation process is discussed.     

Music therapy in palliative care.

Initial observations regarding the use of music therapy at one hospital in the palliative care of patients with advanced malignant disease are presented. In the hands of a trained music therapist, music has proven to be a potent tool for improving the quality of life. The diversity of its potential is particularly suited to the deversity of the challenges - physical, psychosocial and spiritual - that these patients present.     

Functional distribution and further characterization of human endothelial ligand for cellular L-selectin.

In the present study we examine the functional distribution of the human endothelial L-selectin ligand, which determines the sites of extravasation of L-selectin-positive cells. A murine cell line transfected with human L-selectin adhered preferentially to the high endothelial venules (HEV) of human peripheral lymph nodes compared to the HEV of mucosal lymphoid tissues (mean of 0.83 compared to a mean of 0.07 cells per HEV respectively). In addition, an antibody against L-selectin differentially inhibited the adhesion of human lymphocytes to peripheral lymphoid tissue versus mucosal lymphoid tissue HEV (mean 41 and 5% inhibition respectively). Although both sulfoglucuronyl-containing glycolipids and sialyl-Lewis X have been proposed as endothelial ligands for L-selectin, an antibody against the former did not bind to peripheral lymph node endothelium, and an antibody against the latter did not block adhesion of L-selectin-expressing cells. The enzyme O-sialoglycoprotein endopeptidase caused up to an 84% reduction in L-selectin-dependent binding, indicating that sialylated glycoproteins containing O-linked glycans are essential for a large majority of adhesion via L-selectin.     

Morphogenetic pattern formation during ascidian notochord formation is regulative and highly robust.

The ascidian notochord forms through simultaneous invagination and convergent extension of a monolayer epithelial plate. Here we combine micromanipulation with time lapse and confocal microscopy to examine how notochord-intrinsic morphogenetic behaviors and interactions with surrounding tissues, determine these global patterns of movement. We show that notochord rudiments isolated at the 64-cell stage divide and become motile with normal timing; but, in the absence of interactions with non-notochordal tissues, they neither invaginate nor converge and extend. We find that notochord formation is robust in the sense that no particular neighboring tissue is required for notochord formation. Basal contact with either neural plate or anterior endoderm/lateral mesenchyme or posterior mesoderm are each alone sufficient to ensure that the notochord plate forms and extends a cylindrical rod. Surprisingly, the axis of convergent extension depends on the specific tissues that contact the notochord, as do other patterns of cell shape change, movement and tissue deformation that accompany notochord formation. We characterize one case in detail, namely, embryos lacking neural plates, in which a normal notochord forms but by an entirely different trajectory. Our results show ascidian notochord formation to be regulative in a fashion and to a degree never before appreciated. They suggest this regulative behavior depends on a complex interplay between morphogenetic tendencies intrinsic to the notochord plate and instructive and permissive interactions with surrounding tissues. We discuss mechanisms that could account for these data and what they imply about notochord morphogenesis and its evolution within the chordate phylum.     

Derivation of Risk Management Criteria for Chemicals of Unknown Toxic Potency at Contaminated Sites

Environmental investigations of former industrial sites often detect the presence of chemicals for which no soil criteria exist and for which regulatory agencies have not derived estimates of toxic potency. This poses a considerable problem for making informed risk management decisions involving sites where such chemicals are present. As a result, a methodology has been developed for making risk-based decisions for chemicals of unknown toxic potency in soil at contaminated sites. The method is based on principles and procedures used by the US Food and Drug Administration (USFDA), the US Environmental Protection Agency (USEPA) and the Canadian Council of Ministers of the Environment (CCME). After analyzing the data on hundreds of carcinogenic and non-carcinogenic substances, the USFDA and other leading researchers have concluded that, if no toxicological data is available on a chemical, exposures less than 1.5?µg/person/day (i.e., 0.02?µg/kg body weight/day) are unlikely to result in appreciable health risks even if the substance was later found to be a carcinogen. To develop maximum soil concentrations that will be protective of human health (i.e., Risk Management Criteria or RMC), the above exposure limit of 0.02?µg/kg body weight/day has been assumed to be protective of risks from exposure to chemicals lacking toxicological data. Using a stochastic risk assessment model for estimating exposures to chemicals from contaminated sites, our analyses indicate that a soil concentration of 2?µg/g would be protective of human health for land uses that include residential, commercial, and industrial development provided no major indirect pathways exist at the site. If indirect pathways exists (e.g., vapor infiltration of soil gases, uptake of chemicals into garden produce, etc.), alternate RMC could be developed, that include such indirect pathways, using the methodology provided in this paper. Used by experienced risk assessors, the approach is a scientifically defensible screening method that will preclude many chemicals from unnecessary evaluation, while allowing risk assessors to focus efforts on chemicals of greater concern and make informed risk management decisions.     

Estimation of protein sulfhydryl groups with 5,5'-(35S) dithio-bis(2-nitrobenzoate)

³⁵S-Labeled dithio-bis(2-nitrobenzoate) has been prepared and its usefulness for the measurement of protein sulfhydryl groups has been investigated. The technique is at least 20 times more sensitive than when dithio-bis(2-nitrobenzoate) is used colorimetrically. The adduct formed between the reagent and sulfhydryl groups is stable between pH 3–9 which should enable use of the reagent for the tagging of sulfhydryl-containing proteins in a wide variety of biochemical procedures. Removal of the adduct from a protein is achieved simply by treatment with a thiol reagent, with concomitant restoration of enzymic activity of the protein. The procedure has been successfully tested on alcohol dehydrogenase and mammalian ribosomes.     

Uptake of glutamate by Vibrio cholerae in media of low and high osmolarity, and in seawater

Uptake of glutamate by Vibrio cholerae was investigated in media of different osmotic pressure. Accumulation was higher at high osmotic pressures and most accumulation was within the first 6 min of incubation. Greater degradation of glutamate by cells incubated at low osmolarity was insufficient to account for the difference in accumulation levels. The results suggest the glutamate uptake may be important in adaptation of V. cholerae to high salinities such as encountered in estuarine and marine environments.