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PubMed1977

Characterization of functionally distinct lymphoid and myeloid cells from human blood and bone marrow. II. Separation by velocity sedimentation.

Velocity sedimentation in a zonal rotor using gradients of uniform osmolarity was used to separate leukocyte subpopulations from human blood and bone marrow. The separations were performed at high sedimentation rates having the advantage of rapidity over conventional unit gravity separations. Myeloid stem cells (CFU-C) and cells reactive with phytohemagglutinin (PHA), Concanavalin A (Con A), and in mixed leukocyte culture (MLC) were separated and their sedimentation profiles obtained. CFU-C sedimented ahead of lymphoid cells and behind mature myeloid elements. Two distinct marrow subpopulations separated by velocity sedimentation were consistently stimulated by Con A, and variably stimulated by PHA and MLC reactions. Both large cells (predominantly myeloid) and small cells (predominantly lymphoid) from bone marrow were stimulated by Con A in [3H]thymidine incorporation assays. When separated subpopulations showing stimulation by Con A were mixed, inhibition of [3H]thymidine incorporation resulted.

Blood Cells↗
PubMed1979

Organochlorine insecticides and PCB in the sediments of Lake Huron (1969) and Georgian Bay and North Channel (1973).

Surficial and core sediments were collected from the main body of Lake Huron in 1969 and Georgian Bay and North Channel in 1973. These were analysed for organochlorine insecticides and PCB. Residues of organochlorines were higher in the twelve depositional basins in Lake Huron and Georgian Bay than in sediment in the non-depositional zones. PCB was present at similar concentrations to sigma DDT; with mean levels of 13 and 10 ng/g for PCB and sigma DDT in the main body of Lake Huron and 11 and 5 ng/g for PCB and sibma DDT in Georgian Bay. Residues of PCB varied from 9--33 ng/g in the 12 basins. With respect to sigma DDT, both parent DDT and its two metabolites were present in sediment at a mean residue of 7.8 ng/g for the whole lake. The main lake had residues of 10.2 ng/g while Georgian Bay had 5.8 ng/g and North Channel 4.1 ng/g in keeping with use pattenrs since 1943. HEOD was present in only 5.7% of sediment samples from main Lake Huron, 30% from Georgian Bay and 15% from North Channel. The highest residues 1.7 ng/g occurred in the North Channel. No chlordane was detected, however, heptachlor epoxide was identified in 8.5% of sediments collected in Lake Huron, 23% from Georgian Bay and 14% in North Channel. Endosulfan appeared in 4% of samples from both Georgian Bay and North Channel.

Canada↗
PubMed1978

Localization of the site of urinary tract infections by means of antibody-coated bacteria in the urinary sediments.

A prospective study was conducted in children with urinary tract infections to examine the relationship between the presence of antibody-coated bacteria in the urinary sediments and the presence of pyelonephritis. The site of the urinary tract infection was localized as upper or lower urinary tract by the bladder washout technique. Antibody-coated bacteria in the urinary sediments were identified by an immunofluorescent method. In 4 of 12 children with upper urinary tract infections, less than 1% of the bacteria in the urinary sediments were antibody coated. In 10 of 35 children with lower urinary tract infections more than 50% of the bacteria in the urinary sediments were antibody coated. These studies show no significant correlation between the presence of antibody-coated bacteria in urinary sediments and infection of the upper urinary tract.

Adolescent↗
PubMed1977

Dry weight, triglycerides content and glucose-6-phosphatase activity of hepatocytes separated by sedimentation.

1) Isolated rat-hepatocytes were subfractionated by isopycnic or velocity sedimentation, and dry mass, triglycerides content and Glucose-6-Phosphatase activity of different subpopulations were determined. 2) Distribution of the cell dry masses and dry mass per average cell were substantially similar in all the fractions separated by isopycnic sedimentation. Velocity sedimentation allowed a satisfactory separation of cells of different dry mass. 3) Triglycerides content and Glucose-6-Phosphatase activity of cells of the different fractions obtained by isopycnic sedimentation showed no statistically significant difference. Subpopulations fractionated by velocity sedimentation differed in both triglycerides content and Glucose-6-Phosphatase activity, which were substantially parallel to the cell dry mass.

Animals↗
PubMed1978

Sedimentation characteristics of the scrapie agent from murine spleen and brain.

Sedimentation profiles of the scrapie agent in extracts of murine spleen and brain were determined by analytical differential centrifugation. Infectivity profiles of the agent from the two tissues were similar. Sedimentation of the agent was not substantially altered by detergent treatment with sodium deoxycholate. In the presence of detergent, centrifugation at an omega2t value of 3.0 x 1010 rad2/s in a fixed-angle rotor sedimented 90% of the agent. Comparative studies with radioisotopically labeled Simian virus 40 showed that centrifugation at an omega2t value of 1.6 x 10(10) rad2/s removed 90% of the virions. The sedimentation profile of the scrapie agent was similar to that observed for cellular ribosomal RNA. Heating infectious extracts of spleen to 80 degrees C for 30 min resulted in the destruction of 95% of the RNA while sedimentation of the scrapie agent was unchanged. These studies establish a limited range of particle sizes for the scrapie agent.

Animals↗
PubMed1977

Sedimentation properties of the scrapie agent.

The sedimentation behavior of the scrapie agent in homogenates of spleen from infected mice has been determined. Approximately 90% of the scrapie agent was sedimented at an omega2t value of 3 X 10(10) rad2/sec in a fixed-angle rotor. Sedimentation of the agent was not substantially affected by sonication or by treatment with the detergent sodium deoxycholate. The sedimentation profiles of the scrapie agent were similar to those observed for free polyribosomes, but differed from those exhibited by five other subcellular markers. Comparative studies showed that the sedimentation profiles of subcellular markers in spleen suspensions from mice infected with scrapie did not differ from uninoculated controls. These studies suggest that the scrapie agent is a discrete infectious particle which should be separable from cellular membranes.

Animals↗
PubMed1978

Sedimentation cytology in central nervous system leukaemia with a new simple apparatus.

The clinical usefulness of a newly devised, simple apparatus, the Nishimura fluid sedimentation chamber, is described. The latter consists of an open-ended plastic tube, an absorbent paper with a central hole, and a glass slide which are stuck to each other with double-sided plastic adhesive tape with a central hole of equal size. The chamber is set to allow 0.5 ml of cerebrospinal fluid (CSF) to be absorbed in 15--20 min. The principle of the sedimentation relies on the absorption of the fluid portion into the absorbent paper with the cellular elements sedimented onto the glass slide. Thin stationery paper is used as the absorbent material which controls sedimentation speed and minimizes cellular loss. 190 CSF specimens obtained from 28 patients with acute leukaemia were processed by this method. The results obtained showed that sedimentation using this simple chamber is quite reliable for the recognition of small number of abnormal cells in the CSF producing little or no noticeable cellular damage. This technique may be applied to other clinical situations in which fluid cytology is useful in establishing a diagnosis or in evaluating clinical progress.

Acute Disease↗
PubMed1979

Influence of pH, salinity, and organic matter on the adsorption of enteric viruses to estuarine sediment.

This study was designed to determine the degree of adsorption of enteric viruses to marine sediment and factors controlling this association. Adsorption and elution characteristics of several enteroviruses and one rotavirus to estuarine sediments were studied under varying conditions of pH, salinity, and presence of soluble organics. Greater than 99% of the added poliovirus type 1 (LSc), coxsackievirus type B3 (Nancy), echovirus type 7 (Wallace), and rotavirus (SA-11) adsorbed to sediment. Echovirus 1 (Farouk) and a recent isolate typed as coxsackievirus B4 adsorbed significantly less than poliovirus 1 under similar conditions of varying salinity and pH. The presence of soluble organic matter, in the form of secondary sewage effluent or humic acid, did not affect these patterns of adsorption. Only echovirus 1 (Farouk) desorbed when the pH or salinity was altered and then only to a small extent. Three recent isolates of echovirus 1 and echovirus 29 (strain JV-10) also demonstrated varying amounts of adsorption to sediment. These data indicate that enteric viruses can become readily associated with sediment in the estuarine environment and that this association may play a major role in their hydrotransportation and survival.

Adsorption↗
PubMed1976

Intracellular distribution and sedimentation properties of virus-specific RNA in two clones of BHK cells transformed by polyoma virus.

The virus-specific RNA in two independently derived clones of polyoma virus-transformed hamster cells was studied by hybridizing labeled RNA, with excess purified polyoma DNA, immoblized on filters. In one clone (PyBHK1), less than 25% of the total labeled virus-specific RNA was found in the cytoplasm, irrespective of the labeling time. In the other clone (PyBHK2), it was estimated that 39% of the total virus-specific RNA was present inthe cytoplasm after labeling for 3 h. Both the proportion of radioactive label incorporated into virus-specific RNA and the sedimentation pattern of total virus-specific RNA differed markedly between PyBHK and PyBHK2. Most of the virus-specific RNA of PyBHK1 sedimented in the range 25S-35S, whereas a prominent 18S component was present in PyBHK2. Most of the cytoplasmic virus-specific RNA in both clones sedimented at 18S-19S. The sedimentation patterns of virus-specific RNA from whole cells and from washed nuclei of PyBHK1 were closely similar: it was estimated from sedimentation analysis in dimethyl sulfoxide that the molecular weight of 50% of this RNA was within the range 1.1 X10(6) to 2.9 X 10(6). These results, demonstrating the accumulation of virus-specific RNA within the nucleus in at least one virus-transformed cell line, indicate that the large virus-specific RNA previously described in the nuclei of transformed cells may not have represented precursors of virus-specific mRNA.

Cell Line↗
PubMed1976

Biodegradation of petroleum by Chesapeake Bay sediment bacteria.

Chesapeake Bay sediment bacteria from oil-contaminated and oil-free environments were compared for their ability to utilize a South Louisiana crude oil. Preferential solubility, column chromatography, gas-liquid chromatography, and computerized mass spectrometry were used to provide new and useful information regarding biodegradation of fractions and components of the crude oil. Vibrio, Pseudomonas, and Acinetobacter spp. were isolated from the culture inoculated with oil-contaminated sediment, whereas coryneforms and Pseudomonas spp. were isolated from the culture inoculated with oil-free sediment. Microorganisms from the oil-free sediment produced greater quantities of polar n-pentane-insoluble components (asphaltenes) after degradation, whereas microorganisms from the oil-contaminated sediments provided greater degradation of saturated and aromatic hydrocarbons.

Acinetobacter↗
PubMed1978

Antibody-coated bacteria in the urinary sediment of rats with experimental pyelonephritis.

The appearance of antibody-coated bacteria in urinary sediment has been evaluated in rats with experimental pyelonephritis. At day 7 after induction of pyelonephritis, 9 out of 11 rats demonstrated antibody-coated bacteria in the urinary sediment. The other 2 rats never had a positive urinary sediment. Following removal of the pyelonephritic kidneys, antibody-coated bacteria disappeared in 7 of 9 previously positive rats. In the 2 rats which continued to show antibody-coated bacteria, the infecting organisms were found in the remaining kidney. The 2 rats which never developed antibody-coated bacteria in urinary sediment had a higher then normal serum antibody titer, and 1 rat with antibody-coated bacteria showed a normal serum antibody titer. It is concluded that although the search for antibody-coated bacteria in urinary sediment is a very useful technique, its negativity does not exclude upper urinary tract involvement; in the case of renal parenchymal infection, immunity is not the same at the systemic and the local site.

Animals↗
PubMed1978

Effects of phenobarbital and 3-methylcholanthrene pretreatment on size, sedimentation velocity, and mixed function oxygenase activity of rat hepatocytes.

SVA was used to separate liver cells from rats pretreated for 3 days with PB (40 mg/kg intraperitoneally, twice daily) or 3-MC (50 MG/KG AS A SINGLE INJECTIOn). Twelve fractions of cells were collected with s's ranging from 97 mm/hr (fraction1) to 25 mm/hr (fraction 12). Cells in each fraction were sized and counted electronically. PB caused the average volume of the largest cells recovered (fraction 1) to increase to 16, 725 micrometer3 from 10,500 micrometer3 previously reported in untreated animals. The number of cells recovered in the fast-sedimenting fractions (1 to 6) also increased, but there was only a small rise in the average model volume of hepatocytes determined prior to SVA. In cell suspensions analyzed after PB treatment no evidence was found for a discontinuity in the distribution of density-volume characteristics as previously described in hepatocytes from untreated rats. As expected, prior to sedimentation analysis, hepatocyte suspensions from rats treated with PB contained increased cytochrome P-450. The average ratio (n = 4) of P-450 in separated to unseparated cells ranged from 2.75 (fractions 1 + 2) to 0.38 (fractions 11 + 12), giving a 6.8-fold range in quantity of cytochrome per cell. Over this range, cell volume increased 4.2-fold, indicating a gradient in P-450 concentration similar to that previously reported to exist in cells from untreated rat liver. The gradient for AHH activity was 6.7-fold, suggesting that activity of MFO's paralleled the increase in cytochrome concentration. 3-MC pretreatment caused no significant increase in either size or number of cells in the fast-sedimenting fractions, but the discontinuity in density-volume characteristics which distinguished fast and slow sedimenting cells of untreated rats became marked. Furthermore, both the size and number of cells recovered in fractions 7 to 11 (which include the modal peal volume of unseparated hepatocytes) were increased. The gradient of P-450 (OR P1-450) was less steep in 3-MC-treated cells with pooled fractions 1 and 2 containing an average of 4.62 times as much cytochrome as fractions 11 and 12. The gradient for AHH was 4.29 times. The range of cell volume was 3.3-fold over this range. Additional experimental work was performed to determine whether P-450, AHH, or NADPH cytochrome c reductase exhibited differences in activity or concentration per unit of cell volume between cells on either side of fraction 7 where discontinuity had been noted. Each variable was expressed per unit of cell volume in fractions 5 and 9; ratios were compared but were indistinguishable from unity. It was concluded that induction of MFO activity had occurred equally in both populations of cells. Densities were calculated from s and cell volume. Noticeable loss of density followed PB treatment in cells of all sizes. 3-MC had less of an effect on density but enhanced the increment in density observed at fraction 7, which corresponds to the division between cells with distinct sedimentation characteristics...

Animals↗
PubMed1979

Separation of erythroid progenitor cells in mouse bone marrow by isokinetic-gradient sedimentation.

Isokinetic-gradient sedimentation employing a shallow linear gradient of Ficoll in tissue culture medium was used to isolate erythroid progenitor cells (CFU-e) from mouse bone marrow. Following gradient sedimentation, 34% of the total nucleated cells and 48% of the CFU-e applied to the gradient were recovered, and three distinct modal populations of CFU-e could be distinguished. The slowest-migrating population did not require exposure to exogenous erythropoietin in order to form erythroid colonies in vitro. The other two modal populations of CFU-e required exposure to exogenous erythropoietin for differentiation. One of these, constituting 64% of the hormone-dependent CFU-e recovered, migrated with the bulk of the marrow cells, whereas the other migrated ahead of the bulk of the marrow cells. This latter population, which contained 34% of the CFU-e, was recovered with 11% of the marrow cells, representing a twofold to threefold enrichment. BFU-e migrated more slowly than the erythropoietin-dependent CFU-e. Resedimentation studies suggested that the two erythropoietin-dependent CFU-e populations were distinct modal populations. When cells from the fastest-migrating population of erythropoietin-dependent CFU-e were cocultured with unseparated marrow cells, a further twofold to threefold enhancement of erythroid colony formation was obtained. Comparison of isokinetic-gradient sedimentation with discontinuous and continuous albumin density-gradient sedimentation revealed that isokinetic-gradient sedimentation was a more efficient method than the former and a more rapid method than the latter for isolating CFU-e from mouse bone marrow.

Animals↗
PubMed1979

[Experimental study of vibrio parahaemolyticus (biotype 2) transfer from water and sediments to benthic marine food chain organisms].

Transfer of Vibrio parahaemolyticus (biotype 2) from sediments to water and from water to benthic marine organisms was studied experimentally using a streptomycin-resistant strain. Transmission by trophic pathways was also studied using reconstituted marine food chains (Mytilus edulis, Nereis diversicolor, Carcinus maenas, Scorpaena porcus, Mus musculus). Water colonization by sediments could be observed only at temperatures above 16 degrees C. Sediments could well constitute a disseminating reservoir for these germs, their cycle in water being dependent of the cycle followed in the sediments. Contamination of animal organisms is essentially effected by a direct mean, either water or sediments; transfer by trophic pathways being negligible. Infection of land consumers (mice) is linked quantitatively to the nature of the last marine organism of the food chain since bacteria can flourish in the digestive tract of certain animals (Carcinus maenas).

Animals↗
PubMed1978

Characterization of mouse fetal liver granulocyte-macrophage colony-forming cells using velocity sedimentation.

Fetal liver cells from CBA mice were separated by velocity sedimentation to determine the distribution and properties of granulocyte-macrophage colony-forming cells (GM-CFC). Twelve-day fetal liver GM-CFC separated into two peaks (s=7.7 mm/h, s=9.4 mm/h) in cultures stimulated by mouse lung conditioned medium (GM-CSF MLCM) or human urine (GM-CSFHU). Because fetal liver GM-CFC are of relatively light bouyant density, this finding indicates that fetal liver GM-CFC are much larger than corresponding cell in adult bone marrow (s=4.5 mm/h). By 18 days gestation more slowly sedimenting colony-forming cells were present (s=4.7 mm/h, s=6.0 mm/h). At all gestational ages, the most rapidly sedimenting (larger) colony-forming cells were more responsive to stimulation by GM-CSF MLCM than the smaller (slowly sedimenting) cells. Unlike the situation with adult marrow cells, velocity sedimentation achieved no segregation of GM-CFC according to the morphological type of colony produced.

Animals↗
PubMed1977

Dehydrogenase activity in the bottom sediments of three lakes.

Dehydrogenase activity was studied in the sediments of three lakes of the Iława Lakeland (Jeziorak, Mały Jeziorak and Jasne) with reference to the type of lake, the season of the year, the situation of the sediments, pH and temperature. The highest activity was found in the sediments of the eutrophic lake Jeziorak sampled during the summer from places in the vicinity of a village. The lowest activity was exhibited by sediments sampled in the autumn in the lakes Jasne and Mały Jeziorak. No correlation was found between dehydrogenase activity and the number of bacteria in the sediments. The highest dehydrogenase activity was found at 20 degrees and pH 7.0 and 8.0--8.25.

Bacteria↗
PubMed1977

Heterogeneity in human neutrophil, macrophage and eosinophil progenitor cells demonstrated by velocity sedimentation separation.

Progenitor cells of neutrophils, monocyte-macrophages, and eosinophils in human marrow were enumerated in agar cultures stimulated by placental conditioned medium or white cell underlayers. Fractionation of marrow populations by velocity sedimentation showed that the profiles of neutrophil and macrophage colony-forming cells shifted from a peak of 8-9 mm/hr in 7-day cultures to a peak of 6-7 mm/hr in 14-day cultures. This shift was due to degeneration of some early colonies formed by rapidly sedimenting cells and the delayed formation of colonies by slowly sedimenting cells. Eosinophil colony formation was delayed until the second week of incubation. Further evidence of heterogeneity was the observation that rapidly sedimenting colony forming cells were more responsive to stimulation than more slowly sedimenting cells. In the macrophage and eosinophil populations, cluster-forming cells were partially segregatable form colony-forming cells. The observed heterogeneity was similar to the described previously in the mouse and suggests that separate subpopulations of progenitor cells may exist within each hemopoietic family that could possibly give rise to functionally different progeny.

Cell Separation↗
PubMed2025

Tracking microplastic contamination across seasons in a freshwater reservoir: Evidence from surface water, sediments, and fishes.

Microplastics (MPs) are prevalent contaminants in aquatic environments, posing substantial ecological and health risks. These particles migrate within the different layers of aquatic bodies with time and affect the respective biota. Thus, to get an in-depth understanding of the particles, this current study investigated the seasonal distribution, morphological and chemical characteristics, along with potential ecological and human health impacts in the samples including surface water, sediment, and fish from a drinking water supplying reservoir in eastern India. Across three different seasons, pre-monsoon, monsoon, and post-monsoon samples were collected using optimized methods. Results revealed distinct seasonal trends: MP abundance in surface water peaked during the monsoon (mean: 1.15 MPs/L), while sediment showed the highest concentrations in the pre-monsoon (mean: 596 MPs/kg), indicating temporal accumulation dynamics influenced by runoff, hydrodynamics, and sedimentation. Fish gut analysis confirmed ingestion of MPs across five species, with concentrations ranging from 26 to 100 MPs/kg, depending on feeding habits. The most dominant MP type were fragments, followed by fibers, films, and beads. Polymer analysis via µFTIR identified polyethylene, polypropylene, and polyvinyl chloride as prevalent, with hazard assessments (i.e., Polymer Hazard Index (PHI)) indicating medium to very high ecological risks. Heavy metal association was more dominant in the MPs isolated from sediments than the waterborne MPs. Pollution Load Index (PLI) values were > 1 in most seasons, confirming contamination. Health risk analysis suggested potential exposure through both drinking water and fish consumption. This study emphasizes the need for seasonal monitoring, improved waste management, and mitigation strategies to address MP pollution in freshwater ecosystems.

Microplastics↗