Biophysical responses of red cell-membrane systems to very low concentrations of inorganic mercury

Changes in red blood cellsize, deformability, andosmotic fragility are indicators of altered condition and/or altered regulatory processes at the whole cell and membrane levels. An agent, such as HgCl₂, that brings about specific changes of this kind can therefore serve as a selective probe of such cell condition and regulatory state. Conversely, for a health-threatening agent “active” in this way, the cell-membrane responses serve to clarify the more fundamental bases of its toxicity, as well as to permit identification and characterization of its early and low-level actions on living systems. Taking advantage of recent advances in the technique of “resistive pulse spectroscopy,” we present a coordinated study of these three interrelated biophysical properties for the interactions of HgCl₂ with human red cells. We thereby are able to extend previous studies of this kind into domains of shorter time (instantaneous exposures), lower level exposures (down to 10⁻⁹ M, well below the level of acute human toxicity), as well as to additional kinds of responses (e.g., “dynamic osmotic hemolysis”). For conditions ranging from 10⁻⁴ to 10⁻⁹ M in HgCl₂, for instantaneous to 90-min-incubated exposures, for medium osmolarities from 120 to 300, the matrix of observed cell responses includes relative swelling as well as shrinkage, changes in deformability, and both enhancement of and protection against osmotic hemolysis. Some unexpected short-term effects of time and temperature of storage of blood cell stock samples, with respect to increasing and decreasing osmotic fragility, are also reported. These apparently disparate results are interpreted in terms of mercury interactions with cell and membrane SH groups, and a reasonable rationale is presented for most of the responses in terms of disruption of passive and active Na⁺−K⁺, gradient controls, plus interactions with cellular proteins.     

Phylogenetic interpretations and ecological potentials of the Mesomycetozoea (Ichthyosporea)

The Mesomycetozoea (Ichthyosporea) includes a group of fungus-like microorganisms which diverged near the animal fungus divide and is basal to the animal lineage in the Opisthokont supergroup. The composition of species within this group has changed considerably since it was reviewed by Mendoza et al. in 2002. There have been many additions to the group as a result of phylogenetic sequencing, and new species have been discovered following examination of digestive tracts of arthropods and other groups of invertebrates. Furthermore, ecological surveys and environmental sampling have revealed a high diversity of Mesomycetozoean phylotypes in marine, freshwater and terrestrial environments. Initially considered a group comprising mainly fish parasites, the Mesomycetozoea are now known to live in association with a wide range of host organisms including invertebrates, fish, amphibians, birds and mammals. Here we review the Mesomycetozoea and examine the broad spectrum of its ecology in the light of recent research based on sequence data, cultivation and in vivo studies.     

Interaction of snake venom cardiotoxin (a membrane-disruptive polypeptide) with human erythrocytes

The action of 7.2 µM cardiotoxin on 0.25% human erythrocytes in a plasma extender solution was studied by the interaction of toxin with intact red blood cells and subsequent hemolysis of the cells. The binding of toxin to cells was completed within 10 min, whereas the membrane rigidity was weakened in a non-lytic period for about 25 min. The toxin molecules bound almost exclusively to the membrane. The bound toxin could not be liberated with either 0.5% Triton X-100 or 0.1 N NaOH. The degree of binding was slightly reduced in the presence of 10 mM mono- and divalent inorganic salts. The action of toxin might weaken the in situ association of several proteins that are linked with band 3 protein of the membrane, thus making the cells fragile and altering the shape of the cell to a smooth sphere.     

绵羊红细胞分离猪的T细胞方法

参照人的T细胞分离方法用于猪T细胞分离,经二次绵羊红细胞分离的T细胞,用PHA从功能上鉴定获得纯度很高的T细胞,能进一步用于有关T细胞方面研究。     

From stem cells to red blood cells:how far away from the clinical application?

The generation of red blood cells(RBCs)from stem cells provides a solution for deficiencies in blood transfusion.Currently,primary hematopoietic stem cells,embryonic stem cells and induced pluripotent stem cells have shown the potential to produce fully mature RBCs.Here,we discuss the advantages,induction protocols,progress and possible clinical applications of stem cells in RBC production.     

Host–parasite interactions and vectors in the barn swallow in relation to climate change

Recent climate change has affected the phenology of numerous species, and such differential changes may affect host–parasite interactions. Using information on vectors (louseflies, mosquitoes, blackflies) and parasites (tropical fowl mite Ornithonyssus bursa, the lousefly Ornithomyia avicularia, a chewing louse Brueelia sp., two species of feather mites Trouessartia crucifera and Trouessartia appendiculata, and two species of blood parasites Leucozytozoon whitworthi and Haemoproteus prognei) of the barn swallow Hirundo rustica collected during 1971–2008, I analyzed temporal changes in emergence and abundance, relationships with climatic conditions, and changes in the fitness impact of parasites on their hosts. Temperature and rainfall during the summer breeding season of the host increased during the study. The intensity of infestation by mites decreased, but increased for the lousefly during 1982–2008. The prevalence of two species of blood parasites increased during 1988–2008. The timing of first mass emergence of mosquitoes and blackflies advanced. These temporal changes in phenology and abundance of parasites and vectors could be linked to changes in temperature, but less so to changes in precipitation. Parasites had fitness consequences for hosts because intensity of the mite and the chewing louse was significantly associated with delayed breeding of the host, while a greater abundance of feather mites was associated with earlier breeding. Reproductive success of the host decreased with increasing abundance of the chewing louse. The temporal decrease in mite abundance was associated with advanced breeding of the host, while the increase in abundance of the lousefly was associated with earlier breeding. Virulence by the tropical fowl mite decreased with increasing temperature, independent of confounding factors. These findings suggest that climate change affects parasite species differently, hence altering the composition of the parasite community, and that climate change causes changes in the virulence of parasites. Because the changing phenology of different species of parasites had both positive and negative effects on their hosts, and because the abundance of some parasites increased, while that of other decreased, there was no consistent temporal change in host fitness during 1971–2008.     

A simple model of a Gyrodactylus population

Experimental populations of 20 Gyrodactylus alexanderi Mizelle &; Kritsky, 1967, on 19 isolated Gasterosteus aculeatus at 15°C increased for 2 weeks to a mean of 61, then decreased in 2 further weeks to a mean of 9. Fish that lost their fluke infestations were refractory to further infestation for about 3 weeks.The chief factors affecting fluke abundance were measured, including reproduction and mortality rates of flukes on fish, rate of shedding by the fish, mortality rates of flukes while off fish, and the rate of reattachment of the flukes. Data on these individual factors were combined to form a simple deterministic model which simulated the population changes on isolated fish. This was later made more realistic by the introduction of a random variable. When the model was tested in a multiple-host situation it predicted results close to those observed experimentally.     

Testing for Starch,Respiring Tissues,and Vascular Bundles: Inquiry-Based Seed and Stem Anatomy Labs

In this article, the authors present lab exercises in which students use the microscope to study cells. After learning how to recognize the parts of a cell, differentiate between types of cells, and identify the nucleus, the students study blood samples from both familiar and “unknown” animals. The activities allow students to work independently and to become more proficient at microscope use. Students also gain a deeper understanding of the more “invisible” world of science.     

Chicken red blood cells as a substrate for direct polymerase chain reaction

A protocol is presented for direct polymerase chain reaction (PCR) amplification of DNA from chicken nucleated red blood cells. Chicken blood in EDTA was found to have a strong inhibitory effect on the PCR. Consequently, PCR using this protocol should be performed only on a narrow range of blood volumes, from 0125 to 025 |ll.     

鸡红细胞融合最适条件的探讨

目的：探讨快速、有效的细胞融合条件.方法：用鸡红细胞为材料,聚乙二醇（Mw=4000）为诱导剂,诱导鸡红细胞融合.结果：鸡红细胞融合的最适温度为39℃,最适时间为15min.结论：在该条件下,同时用Giemsa染液对融合细胞染色,实验观察效果明显.     

Is there a genetic control in man of red blood cell magnesium levels?

Three groups of human subjects underwent successive monthly examinations to determine red blood cell magnesium levels and plasma magnesium, potassium and sodium levels. From these data, a comparison between intra- and inter individual variances showed that red blood cell magnesium had a characteristically high intra individual stability and a wide inter individual variability. This characteristic, which was much more marked than for plasma cholesterol or serum gamma globulin levels, whose high coefficient of heritability has been demonstrated elsewhere, is in good keeping with the hypothesis of genetic control of red blood cell Mg content.     

Rat red blood cell hexokinase purification,properties and age-dependence

Summary Rat erythrocytes, in contrast to red blood cells from other mammals, have been shown to contain only one hexokinase isozymic form identified as type I by chromatographic and kinetic properties. Rat reticulocytes contain 3.6-times the hexokinase activity found in mature erythrocytes but exactly the same isozyme. By a combination of ion-exchange chromatography, dye-ligand chromatography and high-pressure liquid chromatography the rat erythrocyte hexokinase was purified more than 84 000-fold to a specific activity of 143 units/mg protein and shown to be homogeneous by sodium dodecyl sulfate-gel electrophoresis. The native protein showed a molecular weight of 100 000 by gel-filtration and an apparent molecular weight of 98 000 under denaturating conditions in sodium dodecyl sulfate-gel electrophoresis. The isoelectric point was shown to be 6.3 pH units. This data provides evidence of only one form of hexokinase in the erythrocytes of a mammal.     

Changes in glycine and leucine transport during red cell maturation in the rat

Both glycine and leucine transport in rat red blood cells have been studied. The glycine uptake showed two different components, one sodium-dependent and another diffusion-like process. In contrast, leucine uptake was sodium independent. Both, Na⁺-dependent glycine and the overall leucine uptake in red blood cells showed a saturable pattern. Kinetic parameters in reticulocytes were: i) glycine: apparent Km 0.16 mM; V_max 100.2 nmol/ml ICW/min; ii) leucine: apparent Km 2.11 mM; V_max 3.88 mol/ml ICW/min. The erythrocytes kinetic parameters were: i) glycine: apparent Km 0.17 mM; V_max 9.47 nmol/ml ICW/min; leucine; apparent Km 4.77 mM; V_max 7.42 mol/ml ICW/min. The Kd values (sodium independent glycine uptake) were similar in both kind of cells, but the importance of this component in total glycine uptake in erythrocytes was much higher than in reticulocytes. Our results confirm that rat red blood cells have both saturable leucine and Na⁺-dependent glycine uptake, but some important changes occur during cell maturation.     

Changes in alanine and glutamine transport during rat red blood cell maturation

Alanine and glutamine transport have been studied during red blood cell maturation in the rat. Kinetic parameters of Na⁺-dependent L-alanine transport were:K _m 0.43 and 1.88 mM andV _max 158 and 45 nmoles/ml ICW/min for reticulocytes and erythrocytes, respectively. During red cell maturation in the rat there is a loss of capacity and affinity of the system ASC for L-alanine transport. The values for Na⁺-dependent L-glutamine transport in reticulocytes wereK _m 0.51 mM andV _max 157 nmoles/ml ICW/min. On the other hand, a total loss of L-glutamine transport mediated by both N and ASC systems is demonstrated in mature red cells. This seems to indicate that during rat red cell maturation the system N disappears. Furthermore, the system ASC specificity in mature cells changes, and glutamine enters the red cell by non-mediated diffusion processes.     

Incubation temperature affects multiple measures of immunocompetence in young wood ducks (Aix Sponsa)

Parental effects play a vital role in shaping offspring phenotype. In birds, incubation behaviour is a critical parental effect because it influences the early developmental environment and can therefore have lifelong consequences for offspring phenotype. Recent studies that manipulated incubation temperature found effects on hatchling body composition, condition and growth, suggesting that incubation temperature could also affect energetically costly physiological processes of young birds that are important to survival (e.g. immune responses). We artificially incubated wood duck (Aix sponsa) eggs at three biologically relevant temperatures. Following incubation, we used two immunoassays to measure acquired immune responses of ducklings. Ducklings incubated at the lowest temperature had reduced growth, body condition and responses to both of our immune challenges, compared with those from the higher temperatures. Our results show that incubation temperatures can be an important driver of phenotypic variation in avian populations.     

Cationic and anionic amino acid transport studies in rat red blood cells

The transport of L-proline, L-lysine and L-glutamate in rat red blood cells has been studied. L-proline and L-lysine uptake were Na⁺-independent. When the concentration dependence was studied both showed a non-saturable uptake assimilable to a difussion-like process, with high Kd values (0.718 and 0.191 min^–1 for L-proline and L-lysine respectively). Rat red blood cells showed high impermeability to L-glutamate. No sodium dependence was observed and the Kd value was low (0.067 min^–1). Our results show firstly, that rat red blood cells do not have amino acid transport systems for anionic and cationic amino acids and secondly that erythrocytes show no sodium-dependent L-proline transport, and that these cells are very permeable to this amino acid.Abbreviations MeAIB methyl aminoisobutyric acid     

流行性出血热冻干致敏人O型血球的研制及其应用

本文应用致敏的人O型血球研究反向间接血凝(RPHA)和反向间接血凝抑制(RPHI)方法用以检测流行性出血热抗原抗体,并试验成功用pH9.0硼酸盐水制备灭活鼠脑病毒液作为抗原。为抗原制备提供了一种简便的方法。以上RPHA法用于检测组织培养内病毒与用荧光法检测细胞内病毒抗原法结果一致,用RPHI检测病人血清抗体效价,特异性高,敏感性与IFA相同。该致敏血球和抗原是冻干制品,稳定性好、使用方便,是一种代替荧光检测病毒抗原和抗体的良好制品。