scispace - formally typeset
Search or ask a question
Topic

Lysis

About: Lysis is a research topic. Over the lifetime, 6072 publications have been published within this topic receiving 216978 citations.


Papers
More filters
Journal ArticleDOI
TL;DR: For the first time, a novel integrated device for single-cell analysis (iPAD-1) was developed to profile proteins in a single cell within 1 h, and prominent cellular heterogeneity in protein expressive profiling was observed.
Abstract: In our previous work, we have demonstrated an integrated proteome analysis device (iPAD-100) to analyze proteomes from 100 cells. (1) In this work, for the first time, a novel integrated device for single-cell analysis (iPAD-1) was developed to profile proteins in a single cell within 1 h. In the iPAD-1, a selected single cell was directly sucked into a 22 μm i.d. capillary. Then the cell lysis and protein digestion were simultaneously accomplished in the capillary in a 2 nL volume, which could prevent protein loss and excessive dilution. Digestion was accelerated by using elevated temperature with ultrasonication. The whole time of cell treatment was 30 min. After that, single-cell digest peptides were transferred into an LC column directly through a true zero dead volume union, to minimize protein transfer loss. A homemade 22 μm i.d. nano-LC packing column with 3 μm i.d. ESI tip was used in the device to achieve ultrasensitive detection. A 30 min elution program was applied to analysis of the single-cell proteome. Therefore, the total time needed for a single-cell analysis was only 1 h. In an analysis of 10 single HeLa cells, a maximum of 328 proteins were identified in one cell by using an Orbitrap Fusion Tribrid MS instrument, and the detection limit was estimated at around 1.7-170 zmol. Such a sensitivity of the iPAD-1 was ∼120-fold higher than that of our previously developed iPAD-100 system. (1) Prominent cellular heterogeneity in protein expressive profiling was observed. Furthermore, we roughly estimated the phases of the cell cycle of tested HeLa cells by the amount of core histone proteins.

81 citations

Journal ArticleDOI
TL;DR: The effect of heat shock on tumor necrosis factor‐α and ‐β‐mediated cytolysis of WEHI‐164 clone 13 target cells is studied, suggesting that de novo synthesized, heat‐induced proteins protect target cells from TNF‐mediated lysis in heat shock‐treated WEHI cells.
Abstract: Elevated temperatures and a number of other types of stress induce synthesis of a small number of highly conserved proteins, the heat shock proteins, in a wide variety of cells. The structure and regulation of these proteins have been intensively studied but the question of the function of this universal response has remained unanswered. We studied the effect of heat shock on tumor necrosis factor-alpha (TNF-alpha)- and -beta (TNF-beta)-mediated cytolysis of WEHI-164 clone 13 target cells. One hour pretreatment of target cells at 42 degrees C decreased rTNF-alpha-mediated lysis by 65.3%, 50.5% and 44.8% and TNF-beta-mediated lysis by 61.9%, 43.2% and 38.9% at cytokine concentrations of 0.5 ng/ml, 5 ng/ml and 50 ng/ml, respectively, in an 18-h Cr-release assay. The effect was maximal when TNF-alpha was added 1 h after the heat shock and then gradually declined, being almost undetectable after 2 days. This pattern was found to roughly coincide with the kinetics of hsp68, the major heat-induced protein in murine cells. Heat shock treatment had no protective effect when given 1 h after addition of recombinant TNF-alpha. The heat-induced target cell resistance was not associated with decreased binding of recombinant TNF-alpha to its receptor. Inhibition of protein synthesis by cycloheximide diminished this effect by 76% and inhibition of transcription by actinomycin D abolished it completely, suggesting that de novo synthesized heat-induced proteins protect target cells from TNF-mediated lysis in heat shock-treated WEHI cells.

81 citations

Journal ArticleDOI
TL;DR: The cytotoxic response generated by addition of the TNP-proteins to spleen cell cultures was found to be T-cell dependent at the effector phase, as shown by the sensitivity of the lytic phase to absorbed RAMB and complement.
Abstract: Murine spleen cells from normal donors were cultured in vitro with trinitrobenzene sulfonate (TNBS)-conjugated soluble proteins, i.e., bovine gamma globulin (TNP-BGG) or bovine serum albumin (TNP-BSA). Addition of 100 μg of any of these TNP-proteins to the spleen cell cultures led to the generation of cytotoxic T-cell effectors which were H-2-restricted and TNP- specific. The lytic potential of such effectors was comparable to that generated by sensitization with TNBS-modified syngeneic cells, and was restricted to haplotypes shared at the K or K plus I-A, or the D regions of the H-2 complex. Greater effecter cell activity was generated by addition of TNP-BGG against TNBS-modified targets which shared K plus I-A than against modified targets which shared the D region with the responding cells, which suggests that the same immune response genes are involved when the response is generated by the addition of TNP-conjugated soluble proteins or of TNBS- modified cells. H-2-restricted, TNP-specific effecter cells were generated by culturing mouse spleen cells with syngeneic cells which had been preincubated with TNP- BGG or TNP-BSA for 1.5 h. The addition of unconjugated soluble proteins to the cultures did not result in cytotoxic effectors detectable on H-2-matched targets, whether the targets were prepared by modification with TNBS, or by incubation with either the unconjugated or TNP-conjugated proteins. Depletion of phagocytic cells in the tumor preparation by Sephadex G-10 column fractionation before incubation with TNP-BSA had no effect on their lysis by the relevant effector cells. Immunofluorescent staining of tumor target cells with anti-TNP antibodies indicated that TNP could be detected on the tumor cells within 10 rain of incubation with TNP-BSA. The cytotoxic response generated by addition of the TNP-proteins to spleen cell cultures was found to be T-cell dependent at the effector phase, as shown by the sensitivity of the lytic phase to absorbed RAMB and complement. Furthermore, the response did not appear to be attributable to antibody-dependent cellular cytotoxicity. Three mechanisms were considered which could account for the generation of H-2-restricted, TNP-specific, cytotoxic T-cell effectors by the addition of soluble TNP-proteins. These include covalent linkage of activated TNP groups from the soluble proteins to cell surface components, macrophage processing of the soluble conjugates and presentation to the responding lymphocytes in association with H-2-coded self structures, or hydrophobic interaction of the TNP-proteins to cell surfaces. Results obtained from sodium dodecyl sulfate gel patterns indicating that cell-bound TNP was still linked to BSA, and the observation that phagocytic-depleted cells could interact with the soluble TNP-proteins and function as H-2-restricted targets, appear not to favor the first two proposed mechanisms.

81 citations

Journal ArticleDOI
TL;DR: Both DNA and humic acid concentrations during the extraction and purification of DNA from compost were quantified and direct microscopic observation of pre- and post-lysis samples revealed that 95.3+/-2.3% of native cells was lysed.

81 citations

Journal ArticleDOI
TL;DR: The rates of cell lysis and phage release were determined and it was shown that the efficacy of phage infection was optimal with host cells grown and infected at 26 degrees C.
Abstract: The influence of host growth temperature, phase and media, together with the effect of infection temperature on bacteriophage ΦS1 infection of Pseudomonas fluorescens were examined. The rates of cell lysis and phage release were determined and showed that the efficacy of phage infection was optimal with host cells grown and infected at 26 °C. The host physiological state also affected these rates. Infection was dependent on the presence of cell wall proteins with molecular weights of 17.5 ± 1 and 99 ± 5 kDa.

81 citations


Network Information
Related Topics (5)
Cell culture
133.3K papers, 5.3M citations
86% related
Antigen
170.2K papers, 6.9M citations
86% related
DNA
107.1K papers, 4.7M citations
86% related
Immune system
182.8K papers, 7.9M citations
84% related
Gene
211.7K papers, 10.3M citations
83% related
Performance
Metrics
No. of papers in the topic in previous years
YearPapers
2023389
2022607
2021123
2020142
2019139
2018161