Harvest of human bone marrow directly from bone.

Harvest of human bone marrow directly from freshly resected bone provides purer preparations of marrow than can be obtained by the conventional technique of multiple aspirations from the iliac crests. In particular, directly harvested marrow is much less heavily contaminated with peripheral blood lymphocytes, a known source of mature T cells. Because of the possible relevance of these contaminating T cells for cadaveric bone marrow transplants, the best source of human marrow harvested directly from bone has been studied. Human bone marrow was harvested from 46 surgical specimens and 9 cadaveric tissue donors. Vertebral bodies provided the best source of bone marrow with average yields of 3.1 +/- 1.6 X 10(9) cells per vertebra. When entire ilia were removed and processed for marrow, an average of 1.6 +/- 1.0 X 10(9) cells was obtained. Surgically resected ribs yielded lower amounts of marrow with a mean cell number of 3.2 +/- 2.6 X 10(8) per rib. Isolation of bone marrow mononuclear cells from these preparations by density gradient centrifugation resulted in a loss of 45% of the starting cells. Human bone marrow was found to contain 5-6% T cells before gradient separation and these cells were immunologically competent as measured in vitro by responses to mitogens and alloantigens. This technique may be useful in obtaining human bone marrow for both immunologic studies and allogeneic transplantation.

Cultured T cells from patients with T cell chronic lymphocytic leukemia demonstrate a normal phenotype.

Peripheral blood mononuclear cells from eight patients with the rare T-cell form of chronic lymphocytic leukemia were isolated and cultured with Interleukin-2 (IL-2). In all but one case, cultured T-cells (CTC) were established. Various culture conditions were tested for their effectiveness; feeder layers proved valuable for expanding the cultures to large volumes. The CTC remained IL-2 dependent. Analysis of surface determinants on these CTC showed a polyclonal proliferation of T-cells. The distribution of subset markers in the patients' CTC population had completely changed in comparison to the "fresh" peripheral blood cell population but was similar to CTC initiated from healthy donors. Our data suggest that the patients' few contaminating normal T-lymphocytes expanded in culture, while the malignant cells were unresponsive to IL-2. This conclusion is supported by growth characteristics and morphology of the CTC.

Cokeromyces recurvatus infection in a bone marrow transplant recipient.

Diarrhea is common after bone marrow transplants. We report Cokeromyces recurvatus infection in a transplant recipient with diarrhea. Treatment with mystatin was effective.

A method for the covalent attachment of cells to glass slides for use in immunohistochemical assays.

A method for covalently attaching viable cells to glass slides is described. The cells are linked covalently to the slide without the use of drying or other fixation procedures that normally destroy cellular surface antigens. The attachment is permanent, and the slides with attached cells may be cryopreserved. The use of the avidin-biotin-peroxidase procedure with this procedure shows excellent sensitivity comparable to flow cytometry.

Exocytotic shedding and glial uptake of photoreceptor membrane by a salticid spider.

Receptors in the anterior lateral eyes of salticid spiders possess paired rhabdomeres. The tips of the rhabdomeral microvilli lie adjacent to non-pigmented glial processes. Photoreceptor membrane is lost during turnover by a hitherto undescribed process: individual microvilli lengthen at their tips, taper, and are received by corresponding, coated endocytotic pits in the glial membrane. Pits detach as coated vesicles with coherent fragments of microvilli within them, lose their coats, and accumulate in the glial processes as disorderly membranous detritus. Some microvillus membrane disintegrates before local endocytosis, and appears to get into the glial arms distant from the parent rhabdomere by invaginations which are either endocytotic clefts or a tubulo-cisternal system, but whose precise nature is not yet clear. No photoreceptor membrane is lost by pinocytosis into the receptor cytoplasm. Analogies between the behaviour of this system and the phagocytosis of shed vertebrate photoreceptor membrane are briefly discussed.

Photoreceptor membrane breakdown in the spider Dinopis: localisation of acid phosphatases.

The ultrastructural localisation of acid phosphatases (AcPhs) during the normal daily breakdown of rhabdomere membrane in Dinopis has been examined using beta-glycerophosphate and p-nitrophenyl phosphate as substrates. Results are related to the classification of organelles in the receptors given by Blest, Powell and Kao (1978). Weak and infrequent reactions are obtained in multivesicular bodies (mvbs) and multilamellar bodies (mlbs) derived from them. Residual bodies (rbs) begin to react strongly as they lyse. Source of AcPhs is endoplasmic reticulum which has barely differentiated towards the GERL configuration; it becomes reactive as it is incorporated into secondary lysosomes. GERL tubules, Y-bodies and vesicles respond erratically and weakly, and are also incorporated into rbs. No evidence was found for a significant participation of Golgi bodies in these processes, and acid phosphatase cytochemistry fails to reveal a topographical relationship between GERL in these cells and Golgi saccules. Coated vesicle clusters found in the predawn receptive segments are AcPh-negative; this implies that their previous identification as GERL-derived "Nebenkerne" carrying hydrolytic enzymes to newly-formed mvbs (Blest, Kao and Powell, 1978) is dubious. Isolation bodies and autophagic vacuoles enclosing other organelles in pathological receptors give strong reactions while adjacent secondary lysosomes derived from rhabdomere membrane and associated GERL give weak ones. It is concluded that rhabdomere-derived rb lysis is more tightly regulated than other autophagic processes, and it is suggested that a high degree of control is necessary in a receptor which may repeat the autophagy of a large mass of transductive membrane at least 60--100 times in the course of its working life.

Sodium and potassium levels in Australian processed foods.

We analysed 118 foods available in retail outlets in Sydney for their sodium and potassium content. The concentration of sodium in the types of food examined was (from highest to lowest); salami, ham, luncheon meat, corned beef, liverwurst, breakfast cereals, cracker biscuits, frankfurters, cheese, crispbread, bread, sweet biscuits, canned vegetables, cheesecake and frozen vegetables. There were large variations within some of these groups, especially cheese and breakfast cereals. We have summarized data recently published on 250 other foods. The concentration of potassium was highest in bran-based breakfast cereals and lowest in cheese.

Salting of food--a function of hole size and location of shakers.

The establishment of an association between hypertension and the level of sodium in the diet has focused interest in many countries on the amount of salt added to food, with estimates of intake in western countries being in the range 6-14 g per person per day. As a result, many health authorities have advocated a decrease in salt consumption by the population in general, a common suggestion for achieving a meaningful reduction being to limit the amount of salt used at the table. It is generally assumed that salting habits are influenced by taste preference as shaped by socio-cultural determinants. However, from our observations of the rate of discretionary salt usage of over 1,900 people (mainly adults) consuming main meals in public institutions in Sydney, Australia, were suggest that salting is strongly influenced by the physical factor of mode of presentation of salt to the consumer, particularly the hole size of the salt shaker, and is not influenced by demographic variables.

Contributions of foods to sodium in the Australian food supply.

The sodium contributions of various foods in the Australian supply have been calculated by applying recent local food composition data to food availability data and to typical notional Australian diets. Sodium available for consumption from the food supply was 3.00 g per capita per day. Of foods which are salted during processing, the three heaviest contributors of sodium were bread (23 per cent of total available sodium), processed meat and fish (14 per cent), and margarine (8 per cent). Other important contributors were breakfast cereals, biscuits, cheese, butter, potato crisps, dried soups and cakes (including pastries, pies and puddings). Foods identified by health authorities as 'highly salted' and thus prime targets for reduced consumption, provided 33 per cent of total available sodium, less than the 50 per cent contributed by other foods salted during processing. Take-away foods per se did not contribute more sodium than processed foods generally.

Evaluation and need of education programs for community hospital nurses providing neonatal care.

Phase I: Of 204 RN's providing neonatal care in community hospitals in east Georgia and who were examined using a standardized test, those from large hospitals (greater than 1,000 live births/year) performed significantly better than those from medium or small hospitals (P less than .05). Also, there was negative correlation between test scores and years of experience in areas of the hospital other than the nursery and percent of employment time currently spent in areas of the hospital other than the nursery. Phase II: An educational program in neonatal nursing was then provided to 48 RN's by the authors and to 12 RN's by peer-instructors. At the end of the course of instruction a posttest was given. Thirteen RN's who received no formal instruction served as a control group. On the posttest, both experimental groups performed significantly better than the control group (P less than .01), but the group taught by the authors scored significantly higher than the group taught by peers (P less than .05). Educational programs taught by RN's from the community hospital staff are not as effective in regional educational programs as instruction by course by developers.

Detection of platelet antibody using rosette technique with anti-IgG antibody-coated polyacrylamide gel: a preliminary report.

Paraformaldehyde-fixed platelets from normal donors were used for detection of antibody to platelet by in vitro sensitization (indirect method) utilizing rabbit anti-human IgG heavy chain specific antibody-coated polyacrylamide gels (Immunobeads). The sensitized platelets formed rosettes with Immunobeads and the positive rosette count was over 30%, while control showed less than 8% when normal sera were used. This method was also applicable for detecting antibody-sensitized platelets in vivo (direct method) in patients with autoimmune thrombocytopenia. This method is simple, rapid and reproducible for clinical use. Direct and indirect immunofluorescent antibody tests and a blocking test with anti-human serum also supported the results of Immunobead rosetting technique.

Platelet migration inhibition test for platelet antibodies. Parameters and simplified methodology.

A simplified methodology and parameters associated with the performance of a platelet migration inhibition test for the detection of antiplatelet-associated antibodies is described.