We isolated the cortical microtubules (CMTs) from tobacco BY-2 cells to identify their components. By centrifugation of protoplasts homogenized in the presence of taxol, a MT-stabilizing reagent, in a density gradient of Percoll, we obtained membranous vesicles to which MTs forming a sheet-like bundle were attached. Rhodamine-conjugated Ricinus communis agglutinin I (RCA-I), a lectin that bound...

Hematopoietic stem cells form colonies in the spleens of lethally irradiated mice. The number of colonies is indicative of the number of stem cells present in the inoculum. In this article, a technique is presented for effecting a seven-fold enrichment of spleen colony-forming cells. Bone marrow cells are first segregated into "red" and "white" cell populations by centrifugation on Ficoll-Paque...

Stannin (Snn) is a highly conserved, 88-amino acid protein that may mediate the selective toxicity of organotins. Snn is localized in tissues with known sensitivity to trimethyltin (TMT), including the central nervous system, immune system, spleen, kidney and lung. Cells in culture that do not express Snn show considerable resistance to TMT toxicity. In vitro, Snn peptide can bind TMT in a 1:1 ...

Red blood cell (RBC) fractions were studied after separation of whole blood by means of counterflow centrifugation, Percoll column (Pharmacia, Uppsala, Sweden), and a combination of both separation techniques. Mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), mean corpuscular hemoglobin (MCH), and hemoglobin A1c (HbA1c) were measured in each fraction. From the res...

We have undertaken an analysis of semen from HIV infected men with regard to sperm counts and motility, non-spermatozoal cells, and viral nucleic acid. Regression analysis showed that sperm concentration and motility were positively associated with blood CD4 cell count. By contrast, non-spermatozoal cell concentration (round cells) was inversely related to CD4 count. Extracellular HIV RNA was d...

To study neurons or glia-specific mitochondria one needs to isolate these organelles from primary neuronal or astrocytic cell culture. This work provides novel method for isolation of functional and morphologically intact mitochondria from neurons and astrocytes in cell cultures. In the first step, mitochondria are released from cells by disruption of cell membranes using a nitrogen cavitation ...

We analyzed the expression of myelomonocytic-associated antigens on lymphocytes from B cell chronic lymphocytic leukemia (B-Cu) patients. Blood mononuclear cells were depleted of monocytes by one-step Percoll density gradient centrifugation and tested for antigen expression by fluorescent microscopy and flow cytometry. The reactivity of patient lymphocytes was as follows: 26 of 31 were positive...

The separation of the blood cells of Styela rustica (Styelidae, Stolidobranchiae) in discontinuous Percoll gradient showed 4 fractions. The 4th and bottom most fraction contained 90-100% of morula cells. The protein composition of the morula cell fraction revealed on SDS-PAGE showed two major proteins with m.w. 47 and 26 kDa. These proteins were heavily positively charged. Polyclonal antiserums...

Toxoplasma gondii oocysts are environmentally resistant and can infect virtually all warm-blooded hosts, including humans and livestock. Little is known about the biochemical basis for this resistance of oocysts, and mechanism for excystation of T. gondii sporozoites. The objective of the present study was to evaluate different methods (mechanical fragmentation, gradients, flow cytometry) to se...

The association of glucokinase with liver mitochondria has been reported [Danial et al. (2003) BAD and glucokinase reside in a mitochondrial complex that integrates glycolysis and apoptosis. Nature 424, 952-956]. We confirmed association of glucokinase immunoreactivity with rat liver mitochondria using Percoll gradient centrifugation and demonstrated its association with the 68 kDa regulatory p...