Changes in surface-charge density of blood cells after sudden unexpected death

The objective of the investigation was evaluation of postmortem changes of electric charge of human erythrocyte and thrombocyte membranes after sudden unexpected death. The surface charge density values were determined on the basis of the electrophoretic mobility measurements of the cells carried out at various pHs of electrolyte solution. The interactions between both erythrocyte and thrombocyte membranes and electrolyte ions were studied. Values of parameters characterizing the membrane--that is, the total surface concentrations of both acidic and basic groups and their association constants with solution ions--were calculated on the basis of a four-equilibria mathematical model. The model was validated by comparison of these values to experimental data. We established that examined electric properties of the cell membranes are affected by sudden unexpected death. Postmortem processes occurring in the cell membranes can lead to disorders of existing equilibria, which in turn result in changes in values of all the above-mentioned parameters.     

The mitochondrial battlefield and membrane lipids during cell death signalling

This brief review focuses on current issues regarding the biochemical understanding of key reactions in the mitochondrial membrane damage induced by apoptosis signalling, especially in the pathway followed by death receptors in primary tissues. An overview of the emerging role of cardiolipin in apoptosis is also presented, discussing controversial issues and future directions of research. In addition, it is suggested that pro-apoptotic Bid may modulate key processes linking lipid constituents of mitochondrial membranes to lipid re-modelling that are altered early by death receptor signalling.     

Cell Death Induced by Mild Physical Perturbations Could Be Related to Transient Plasma Membrane Modifications

An understanding of membrane destabilization induced by osmotic treatments is important to better control cell survival during biotechnological processes. The effects on the membranes of the yeast Saccharomyces cerevisiae of perturbations similar in intensity (same amount of energy) but differing in the source type (heat, compression and osmotic gradient) were investigated. The anisotropy of the fluorescent probe 1,6-diphenyl-1,3,5-hexatriene was measured before and after each treatment to assess the reversibility of the membrane changes related to each treatment. Except for heat shock at 75°C, changes in membrane fluidity were reversible after the return to initial conditions, showing that two kinds of physical stress can be distinguished regarding the reversibility of membrane changes: high and mild energy stresses. With the application of osmotic gradients, anisotropy was assessed during treatment with five osmotic pressure levels from 30.7 to 95.4 MPa with two different yeast strains and related to the rate of cell death caused by each stress. The exposure of cells to increasing osmotic pressures involved a progressive lowering of membrane anisotropy during lethal perturbations. Osmotic stresses associated with reversible fluidity changes of increasing intensity in the membrane led to proportional death rates and time-dependant cell death of increasing rapidity during the application of the stress. Finally, a hypothesis relating the extent of membrane structural changes to the kinetic of cell death is proposed.     

Study of thymocytes resistance to ionising irradiation in vitro

The influence of ionising irradiation with doses of 0.5, 1, 2 Gy upon the suspension of mice thymocytes was studied. The state of the system of lipid peroxidation-antioxidation activity and structural and functional state of cell membrane was studied. It was found that, 15 minutes' after irradiation POL processes are within the norm. But the content of vitamins A and E decreases (the former after 0.5 Gy, the latter after 1 Gy). Changes in structural and functional state of thymocyte membranes can be observed by using fluorescent probes. Particularly, it turned out that after irradiation accessibility of incorporated proteins increases, the polarity of lipids increases and the lipophility of membrane lipids decreases. These changes can lead to short-time increase in specific functional activity of cells, but they can cause rapid death of cells as well.     

Ultrastructural analysis of interneuronal syncytial perforations

The structural regularities of the organization of interneuronal syncytial cytoplasmic connections between neuronal bodies in gyrus dentatus and CA1 and CA2 (CA is cornu ammonis) of hippocampus, as well as between cell neurites of the caudal mesenteric ganglion were studied by transmission electron microscopy. The syncytial perforations are located only on the base of tight junctions. The perforations have rounded edges corresponding to the fusion edges of perforated membranes of adjacent neurons – or where their edges have a form of thinned plate – a remnant of the tight junction. In the lumen of the perforations, remnants of contact membranes – residual bodies – are revealed. On living neurons in tissue culture, the syncytial connection of two contacting processes of different neurons is found during the death of the body of one of them, but with preservation of viability of its processes that contact with other neurons.     

Mechanics and Dynamics of Bacterial Cell Lysis

Membrane lysis, or rupture, is a cell death pathway in bacteria frequently caused by cell wall-targeting antibiotics. Although previous studies have clarified the biochemical mechanisms of antibiotic action, a physical understanding of the processes leading to lysis remains lacking. Here, we analyze the dynamics of membrane bulging and lysis in Escherichia coli, in which the formation of an initial, partially subtended spherical bulge (“bulging”) after cell wall digestion occurs on a characteristic timescale of 1 s and the growth of the bulge (“swelling”) occurs on a slower characteristic timescale of 100 s. We show that bulging can be energetically favorable due to the relaxation of the entropic and stretching energies of the inner membrane, cell wall, and outer membrane and that the experimentally observed timescales are consistent with model predictions. We then show that swelling is mediated by the enlargement of wall defects, after which cell lysis is consistent with both the inner and outer membranes exceeding characteristic estimates of the yield areal strains of biological membranes. These results contrast biological membrane physics and the physics of thin, rigid shells. They also have implications for cellular morphogenesis and antibiotic discovery across different species of bacteria.     

The Effect of Fatal Carbon Monoxide Poisoning on the Surface Charge of Blood Cells

The objective of this investigation was to evaluate postmortem changes of electric charge of human erythrocytes and thrombocytes after fatal carbon monoxide (CO) poisoning. The surface charge density values were determined on the basis of the electrophoretic mobility measurements of the cells carried out at various pH values of electrolyte solution. The surface charge of erythrocyte membranes after fatal CO poisoning as well as after sudden unexpected death increased compared to the control group in the whole range of experimental pH values. Also, a slight shift of the isoelectric point of erythrocyte membranes to high pH values was observed. The surface charge of thrombocyte membranes after fatal CO poisoning decreased at low pH compared to the control group. However, at high pH, the values increased compared to the control group. The isoelectric point of thrombocyte membranes after fatal CO poisoning was considerably shifted toward low pH values compared to the control group. The observed changes are probably connected with the destruction of blood cell structure.     

鞘脂在植物-真菌互作中的分子调控机制研究进展

鞘脂是细胞生物膜结构的重要组分, 鞘脂及其代谢产物参与许多重要的信号转导过程。在植物-真菌互作中, 植物鞘脂的主要作用是诱导细胞发生程序性死亡; 真菌鞘脂既能引起植物死亡, 也能诱导植物产生抗病性。该文总结了植物和真菌鞘脂的结构及代谢特点, 综述了鞘脂参与调控植物-真菌互作的分子机制研究进展, 并展望了植物-真菌共生关系中鞘脂作用的研究方向。     

鞘脂在植物-真菌互作中的分子调控机制研究进展

鞘脂是细胞生物膜结构的重要组分, 鞘脂及其代谢产物参与许多重要的信号转导过程。在植物-真菌互作中, 植物鞘脂的主要作用是诱导细胞发生程序性死亡; 真菌鞘脂既能引起植物死亡, 也能诱导植物产生抗病性。该文总结了植物和真菌鞘脂的结构及代谢特点, 综述了鞘脂参与调控植物-真菌互作的分子机制研究进展, 并展望了植物-真菌共生关系中鞘脂作用的研究方向。     

Mitochondrial ion channels as therapeutic targets

The study of mitochondrial ion channels changed our perception of these double-wrapped organelles from being just the power house of a cell to the guardian of a cell’s fate. Mitochondria communicate with the cell through these special channels. Most of the time, the message is encoded by ion flow across the mitochondrial outer and inner membranes. Potassium, sodium, calcium, protons, nucleotides, and proteins traverse the mitochondrial membranes in an exquisitely regulated manner to control a myriad of processes, from respiration and mitochondrial morphology to cell proliferation and cell death. This review is an update on both well established and putative mitochondrial channels regarding their composition, function, regulation, and therapeutic potential.     

Src-family kinase signaling,actin-mediated membrane trafficking and organellar dynamics in the control of cell fate: Lessons to be learned from the adenovirus E4orf4 death factor

Evidence has accumulated that there are different modes of regulated cell death, which share overlapping signaling pathways. Cytoskeletal-dependent inter-organellar communication as a result of protein and lipid trafficking in and out of organelles has emerged as a common, key issue in the regulation of cell death modalities. The movement of proteins and lipids between cell compartments is believed to relay death signals in part through modifications of organelles dynamics. Little is known, however, regarding how trafficking is integrated within stress signaling pathways directing organelle-specific remodeling events. In this review, we discuss emerging evidence supporting a role for regulated changes in actin dynamics and intracellular membrane flow. Based on recent findings using the adenovirus E4orf4 death factor as a probing tool to tackle the mechanistic underpinnings that control alternative modes of cell death, we propose the existence of multifunctional platforms at the endosome-Golgi interface regulated by SFK-signaling. These endosomal platforms could be mobilized during cell activation processes to reorganize cellular membranes and promote inter-organelle signaling.     

Ion channnels and transporters in cancer. 5. Ion channels in control of cancer and cell apoptosis

Ion channels contribute to virtually all basic cellular processes, including such crucial ones for maintaining tissue homeostasis as proliferation, differentiation, and apoptosis. The involvement of ion channels in regulation of programmed cell death, or apoptosis, has been known for at least three decades based on observation that classical blockers of ion channels can influence cell death rates, prolonging or shortening cell survival. Identification of the central role of these channels in regulation of cell cycle and apoptosis as well as the recent discovery that the expression of ion channels is not limited solely to the plasma membrane, but may also include membranes of internal compartments, has led researchers to appreciate the pivotal role of ion channels plays in development of cancer. This review focuses on the aspects of programmed cell death influenced by various ion channels and how dysfunctions and misregulations of these channels may affect the development and progression of different cancers.     

Modulation of mitochondrial ion transport by inorganic polyphosphate - essential role in mitochondrial permeability transition pore

Inorganic polyphosphate (polyP) is a biopolymer of phosphoanhydride-linked orthophosphate residues. PolyP is involved in multiple cellular processes including mitochondrial metabolism and cell death. We used artificial membranes and isolated mitochondria to investigate the role of the polyP in mitochondrial ion transport and in activation of PTP. Here, we found that polyP can modify ion permeability of de-energised mitochondrial membranes but not artificial membranes. This permeability was selective for Ba²⁺ and Ca²⁺ but not for other monovalent and bivalent cations and can be blocked by inhibitors of the permeability transition pore – cyclosporine A or ADP. Lower concentrations of polyP modulate calcium dependent permeability transition pore opening. Increase in polyP concentrations and elongation chain length of the polymer causes calcium independent swelling in energized conditions. Physiologically relevant concentrations of inorganic polyP can regulate calcium dependent as well calcium independent mitochondrial permeability transition pore opening. This raises the possibility that cytoplasmic polyP can be an important contributor towards regulation of the cell death.     

Gangliosides as apoptotic signals in ER stress response

Renewed attention has been given lately to gangliosides and to their function as intracellular messengers of the adaptive responses to stress. Gangliosides are vital components of cell membranes; therefore, deleterious consequences can result from changes in their chemical composition and concentration, that is, membrane dynamics and structure can be altered as can the behavior of other membrane proteins. The importance of gangliosides in human health is evident in neurodegenerative diseases associated with defects in their degradation. As key modulators of intracellular calcium flux, gangliosides are involved in cellular processes downstream of calcium signaling. In this review, we focus on the effect of ganglioside accumulation on the endoplasmic reticulum calcium homeostasis and on the integrity of the mitochondrial membranes. We discuss how these events elicit an apoptotic program that ultimately leads to cell death. Owing to interorganelle crosstalk, these events are not necessarily self-contained, and gangliosides may serve as the common factor.     

Partial urethral obstruction enhances NADPH-diaphorase activity in the monkey (Macaca fascicularis) bladder: light and electron microscopic studies

The effect of partially obstructing the urethra on the nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) activity in neurons of the intramural ganglia of the monkey (Macaca fascicularis) bladder was examined by light and electron microscopy. Partial urethral ligation was done in adult male monkeys. The animals were sacrificed 2, 4 weeks after partial urethral obstruction. This was compared to controls (normal and sham operated). Urethral obstructed animals were observed to have increased urinary frequency and decreased urinary flow rate. Two weeks after urethral obstruction, the overall NADPH-d activity in the intramural ganglia of the bladder base was enhanced compared to control animals. The frequency of intensely stained NADPH-d positive neurons was increased compared to the control animals. About one-third of intensely stained NADPH-d positive neurons appeared to undergo degenerative changes. At 4 weeks after urethral obstruction, a wide occurrence of NADPH-d positive neurons in advanced stages of degeneration in the bladder base was observed. Cellular debris was strewn among normal looking ganglion cells and along the nerve processes. The proportion of intensely stained NADPH-d positive neurons was relatively lower than the controls. The total number of NADPH-d positive neurons and the nerve fibres in the entire bladder was significantly reduced when compared to control animals. Electron microscopy showed some NADPH-d activity in intramural ganglion cells in 2 weeks after partial urethral obstruction. NADPH-d reaction product (formazan) was deposited on the membranes of the rough endoplasmic reticulum, and the outer membranes of some mitochondria in the intramural neuron. At 4 weeks after urethral obstruction, NADPH-d was present in the membrane of the mitochondria and some mitochondria appeared swollen with disrupted cristae. Present results show that NADPH-d activity in neurons of the intramural ganglia of the monkey (Macaca fascicularis) urinary bladder was increased after two weeks and reduced after 4 weeks of partial urethral obstruction. It is speculated that the increased NADPH-d activity associated with partial urethral obstruction would lead to neuronal damage and death, which may contribute to detrusor overactivity. However, it warrants further investigation to understand the mechanism of neuronal cell death after partial urethral obstruction.     

Subcellular localization of Photofrin determines the death phenotype of human epidermoid carcinoma A431 cells triggered by photodynamic therapy: when plasma membranes are the main targets

Photodynamic therapy (PDT) is a kind of photochemo-therapeutic treatment that exerts its effect mainly through the induction of cell death. Distinct types of cell death may be elicited by different PDT regimes. In this study, the mechanisms involved in the death of human epidermoid carcinoma A431 cells triggered by PDT with Photofrin (a clinically approved photosensitizer) were characterized. Photofrin distributes dynamically in A431 cells; the plasma membranes and Golgi complex are the main target sites of Photofrin after a brief (3 h) and prolonged (24 h) incubation, respectively. Cells with differentially localized Photofrin displayed distinct death phenotypes in response to PDT. The effects of PDT on cells with plasma membrane-localized Photofrin were further studied in details. Cells stopped proliferating post PDT at Photofrin dose >7 micro g/ml, and at higher dose (28 micro g/ml) plasma membrane disruption and cell swelling were observed immediately after PDT. Dramatic alterations of several important signaling events were detected in A431 cells post Photofrin-PDT, including (i) immediate formation of reactive oxygen species (ROS), (ii) rapid activation of c-Jun N-terminal kinase, (iii) delayed activation of caspase-3 and cleavage of polyADP-ribose polymerase and p21-activated kinase 2, and (iv) loss of mitochondrial membrane potential. Intriguingly, the characteristics of typical apoptosis such as phosphatidylserine externalization and DNA fragmentation were not detected in the cell death process caused by this PDT regime. In conclusion, our results show that when plasma membranes are the main targets, Photofrin-PDT can lead to instant ROS formation and subsequent activation of downstream signaling events similar to those elicited by many apoptotic stimuli, but the damage of plasma membranes renders the death phenotype more necrosis like.     

Characteristics of the radiation-induced changes in the content of sterols and squalene in the lymphoid system tissues and erythrocyte membranes of rats

Ionizing radiation causes considerable changes in the content of sterols and squalene in the lymphoid system tissues and erythrocyte membranes which is in accordance with the concept of high radiosensitivity of haemopoietic tissue. The processes of cholesterol conversion to its oxy-derivatives are increased under the effect of radiation. The content of some lipid components in the lymphoid system tissues and erythrocyte membranes is changed depending on the dose and time after irradiation. There is a relationship between the changes in the sterol composition and in the properties of erythrocyte membranes.     

The therapeutic potential of mitochondrial channels in cancer, ischemia-reperfusion injury, and neurodegeneration

Mitochondria communicate with the rest of the cell through channels located in their inner and outer membranes. Most of the time, the message is encoded by the flow of anions and cations e.g., through VDAC and PTP, respectively. However, proteins are also both imported and exported across the mitochondrial membranes e.g., through TOM and MAC, respectively. Transport through mitochondrial channels is exquisitely regulated and controls a myriad of processes; from energy production to cell death. Here, we examine the role of some of the mitochondrial channels involved in neurodegeneration, ischemia-reperfusion injury and cancer in the context of their potential as therapeutic targets.     

E. coli hemolysin interactions with prokaryotic and eukaryotic cell membranes.

The hemolysin toxin (HlyA) is secreted across both the cytoplasmic and outer membranes of pathogenic Escherichia coli and forms membrane pores in cells of the host immune system, causing cell dysfunction and death. The processes underlying the interaction of HlyA with the bacterial and mammalian cell membranes are remarkable. Secretion of HlyA occurs without a periplasmic intermediate and is directed by an uncleaved C-terminal targetting signal and the HlyB and HlyD translocator proteins, the former being a member of a transporter superfamily central to import and export of a wide range of substrates by prokaryotic and eukaryotic cells. The separate process by which HlyA is targetted to mammalian cell membranes is dependent upon fatty acylation of a non-toxic precursor, proHlyA. This is achieved by a novel mechanism directed by the activator protein HlyC, which binds to an internal proHlyA recognition sequence and provides specificity for the transfer of fatty acid from cellular acyl carrier protein.     

FORMATION OF MYELIN IN THE CENTRAL NERVOUS SYSTEM OF MICE AND RATS, AS STUDIED WITH THE ELECTRON MICROSCOPE

Sections of brain and spinal cord of mice and rats at 1, 3, 5, and 17 days after birth were examined with the electron microscope. In the early stages of myelinization only two to four lamellae surround the axon. These laminae are formed from the plasma membraces of glial processes, and in particular of oligodendroglial processes. In a later stage of myelinization a larger number of flattened glial processes surround the axon with enclosed cytoplasm trapped within some of the membranes. Multiple extensions of the membranes of the flattened glial processes to the lamellae of the myelinated sheath are evident at this stage, with a variable number of membranes within the sheath at various positions along the fiber. Newly formed myelinated sheaths are sometimes larger than their enclosed fibers, extending as a projection of sheath which does not surround axoplasm. Loci are present in which the myelinated sheath is incomplete or interrupted.