Milestones and recent discoveries on cell death mediated by mitochondria and their interactions with biologically active amines.

Mitochondria represent cell "powerhouses," being involved in energy transduction from the electrochemical gradient to ATP synthesis. The morphology of their cell types may change, according to various metabolic processes or osmotic pressure. A new morphology of the inner membrane and mitochondrial cristae, significantly different from the previous one, has been proposed for the inner membrane and mitochondrial cristae, based on the technique of electron tomography.

Spermine cycling in mitochondria is mediated by adenine nucleotide translocase activity: mechanism and pathophysiological implications.

Spermine, besides to be transported in mitochondria by an energy dependent electrophoretic mechanism, can be also released by two different mechanisms. The first one is induced in deenergizing conditions by FCCP or antimycin A and it is mediated by an electroneutral exchange spermine protons. The second one takes place in energizing conditions during the activity of the adenine nucleotide translocase and is mediated by an electroneutral symport mechanism involving the efflux in co-transport of spermine and phosphate and the exchange of exogenous ADP with endogenous ATP.

Effect of various commercial buffers on sperm viability and capacitation.

A wide variety of sperm preparation protocols are currently available for assisted conception. They include density gradient separation and washing methods. Both aim at isolating and capacitating as much motile sperm as possible for subsequent oocyte fertilization.

Bidirectional fluxes of spermine across the mitochondrial membrane.

The polyamine spermine is transported into the mitochondrial matrix by an electrophoretic mechanism having as driving force the negative electrical membrane potential (ΔΨ). The presence of phosphate increases spermine uptake by reducing ΔpH and enhancing ΔΨ. The transport system is a specific uniporter constituted by a protein channel exhibiting two asymmetric energy barriers with the spermine binding site located in the energy well between the two barriers.

Inositol administration reduces oxidative stress in erythrocytes of patients with polycystic ovary syndrome.

Objective: Possibly due to a deficiency of insulin mediators, polycystic ovary syndrome (PCOS) is often associated with insulin resistance (IR) and hyperinsulinemia, likely responsible for an elevated production of reactive oxygen species. We investigated oxidative-related alterations in erythrocytes and anti-inflammatory effects of inositol in women with PCOS before and after treatment with myo-inositol (MYO).

Methods: Twenty-six normal-weight PCOS patients were investigated before and after MYO administration (1200 mg/day for 12 weeks; n=18) or placebo (n=8) by evaluating serum testosterone, serum androstenedione, fasting serum insulin, fasting serum glucose, insulin area under the curve (AUC), and glucose AUC after oral glucose tolerance test and homeostasis model of assessment-IR.

Effect of peroxides on spermine transport in rat brain and liver mitochondria.

The polyamine spermine is transported into the matrix of various types of mitochondria by a specific uniporter system identified as a protein channel. This mechanism is regulated by the membrane potential; other regulatory effectors are unknown. This study analyzes the transport of spermine in the presence of peroxides in both isolated rat liver and brain mitochondria, in order to evaluate the involvement of the redox state in this mechanism, and to compare its effect in both types of mitochondria.

Further characterization of agmatine binding to mitochondrial membranes: involvement of imidazoline I2 receptor.

Agmatine, a divalent diamine with two positive charges at physiological pH, is transported into the matrix of liver mitochondria by an energy-dependent mechanism, the driving force of which is the electrical membrane potential. Its binding to mitochondrial membranes is studied by applying a thermodynamic treatment of ligand-receptor interactions on the analyses of Scatchard and Hill. The presence of two mono-coordinated binding sites S(1) and S(2), with a negative influence of S(2) on S(1), has been demonstrated.

Regulation of membrane band 3 Tyr-phosphorylation by proteolysis of p72(Syk) and possible involvement in senescence process.

Erythrocyte senescence is characterized by exposure of cell surface epitopes on cell membrane proteins leading to immune mediated removal of red blood cells. One mechanism for antigen formation is tyrosine phosphorylation (Tyr-P) of the transmembrane protein band 3 by Syk kinase. Our aim was to test the hypothesis that proteolytic activation of Syk kinase by conversion from 72 kDa (p72(Syk)) to the 36 kDa (p36(Syk)) isoform enhances its phosphorylating activity independently of the association of Syk kinase with the cytoskeleton.

Immunotoxicity in ascidians: antifouling compounds alternative to organotins - II. The case of Diuron and TCMS pyridine.

Using short-term hemocyte cultures of the colonial ascidian Botryllus schlosseri exposed to various sublethal concentrations of Diuron (3-(3,4-diclorophenyl)-1,1-dimethylurea) and TCMS pyridine (2,3,5,6-tetrachloro-4-(metylsulphonyl)pyridine), we evaluated their immunotoxic effects through a series of cytochemical assays previously used for organotin compounds. At concentrations higher than 250 micro M and 10 micro M for Diuron and TCMS pyridine, respectively, both biocides exerted immunosuppressant effects on Botryllus hemocytes, causing i) deep changes in the cytoskeleton that irreversibly affect cell morphology and phagocytosis, ii) induction of DNA damage, iii) leakage of oxidative and hydrolytic enzymes due to membrane alteration. Unlike organotin compounds, Diuron and TCMS pyridine do not inhibit cytochrome-c-oxidase, and only TCMS pyridine triggers oxidative stress.

A comparison between the responses of neutral red and acridine orange: acridine orange should be preferential and alternative to neutral red as a dye for the monitoring of contaminants by means of biological sensors.

The acridine orange (AO) and neutral red (NR) dyes, commonly used as probes to measure the internal pH in acidic vesicles, are compared in this article. The comparison between the two dyes (arising from calculations taking into account their analytical constants) illustrated that the use of AO is preferential to that of NR because the AO response is sensitive over the whole pH range between 4.0 and 7.

Cobalt induces oxidative stress in isolated liver mitochondria responsible for permeability transition and intrinsic apoptosis in hepatocyte primary cultures.

It is well established that cobalt mediates the occurrence of oxidative stress which contributes to cell toxicity and death. However, the mechanisms of these effects are not fully understood. This investigation aimed at establishing if cobalt acts as an inducer of mitochondrial-mediated apoptosis and at clarifying the mechanism of this process.

Toxic effects of new antifouling compounds on tunicate haemocytes I. Sea-nine 211 and chlorothalonil.

After the definitive ban on tin-based antifouling substances, new organic compounds have recently been introduced in antifouling paint formulations, as either principal or booster biocides. In most cases, previous risk assessment of these biocides has been inadequate so that their possible effects on aquatic ecosystems is a matter of great concern. We studied the effects of two new organic biocides often associated in paint formulations, Sea-Nine 211 (4,5 dichloro-2-n-octyl-4-isothiazoline-3-one) and chlorothalonil (2,4,5,6-tetrachloroisophthalonitrile), on haemocytes of the compound ascidian Botryllus schlosseri exposed for 60 min to various concentrations (from 0.

TCMS inhibits ATP synthesis in mitochondria: a systematic analysis of the inhibitory mechanism.

The interactions of the antifouling compound TCMS (2,3,5,6-tetrachloro-4-methylsulphonyl pyridine) with rat liver mitochondria have been investigated. The results indicate that the compound inhibits ATP synthesis. Further investigations regarding the ATP synthesis mechanism suggest that TCMS inhibits succinic dehydrogenase of the mitochondrial respiratory chain.

SHP-1 tyrosine phosphatase in human erythrocytes.

SHP-1 is a SH2-domain containing protein Tyr-phosphatase expressed in hematopoietic cell lines, which is hypothesized to play a negative role in signal transduction. In human erythrocytes, the phospho-Tyr level of proteins, mainly transmembrane band 3, is closely controlled by the antithetic activity of Tyr-protein kinases and phosphatases, resulting in a dephosphorylated state. Only after particular stimuli, as with oxidizing agents, diamide or pervanadate, or thiol alkylating compound, N-ethyl maleimide (NEM), Tyr-phosphorylation of band 3 can be triggered, inhibiting Tyr-phosphatase action and inducing erythrocyte membrane reorganization.

Trialkyllead compounds induce the opening of the MTP pore in rat liver mitochondria.

The interactions of the tributyl, triethyl and trimethyllead compounds with energized mitochondria have been investigated in this paper. It has been shown that the (alkyl)(3)Pb-Cl compounds induce swelling in mitochondria suspended in a sucrose medium. The phenomenon is more marked the higher the lipophilicity and occurs in the following order: (Bu)(3)Pb>(Et)(3)Pb>(Me)(3)Pb.

The interactions of cobalt(II) with mitochondria from rat liver.

The interactions of Co(2+) with mitochondria have been investigated. The results indicate that Co(2+) inhibits ATP synthesis. Further investigations into ATP synthesis mechanisms indicated that inhibition is due to the opening of a transmembrane pore.

Irgarol inhibits the synthesis of ATP in mitochondria from rat liver.

The interactions of Irgarol with rat liver mithocondrial have been investigated. The results indicate that Irgarol inhibits the ATP synthesis. The analysis of the various steps involved in the ATP synthesis suggests that the inhibition is due to the opening of small-size pores.

An in vitro study of the interaction of sea-nine with rat liver mitochondria.

The interactions of the antifouling compound Sea-Nine with rat liver mitochondria have been studied. The results indicate that low doses of this compound inhibit adenosine 5'-triphosphate (ATP) synthesis. Further investigations indicate that ATP synthesis inhibition should be due to an interaction of Sea-Nine with the succinic dehydrogenase in the mitochondrial respiratory chain.

Gold complexes inhibit mitochondrial thioredoxin reductase: consequences on mitochondrial functions.

The effects of gold(I) complexes (auranofin, triethylphosphine gold and aurothiomalate), gold(III) complexes ([Au(2,2'-diethylendiamine)Cl]Cl(2), [(Au(2-(1,1-dimethylbenzyl)-pyridine) (CH(3)COO)(2)], [Au(6-(1,1-dimethylbenzyl)-2,2'-bipyridine)(OH)](PF(6)), [Au(bipy(dmb)-H)(2,6-xylidine)](PF(6))), metal ions (zinc and cadmium acetate) and metal complexes (cisplatin, zinc pyrithione and tributyltin) on mitochondrial thioredoxin reductase and mitochondrial functions have been examined. Both gold(I) and gold(III) complexes are extremely efficient inhibitors of thioredoxin reductase showing IC(50) ranging from 0.020 to 1.

A possible transport mechanism for aluminum in biological membranes.

The transport mechanism of aluminum in lysosomes extracted from rat liver has been investigated in this paper. The experimental evidence supports the hypothesis that aluminum is transported inside lysosomes in the form of an Al(OH)(3) electroneutral compound, the driving force being the internal acidic pH. This mechanism could help to explain the presence of aluminum in cells in many illnesses.

Thallium induces apoptosis in Jurkat cells.

Thallium induces swelling in mitochondria and apoptosis in Jurkat cells. Both the swelling and the apoptosis are inhibited by means of cyclosporine A (CsA), thus suggesting that these phenomena result from the opening of the membrane transition pore (MTP) in mitochondria. Therefore, apoptosis can be explained as due to the opening of the MTP in mitochondria.

Frozen mitochondria as rapid water quality bioassay.

A rapid and relatively low cost bioassay, usable in routine screening water test has been developed modifying the beef heart mitochondria test. In our experiments, mitochondria (FM22) were frozen at -22 degrees C, instead of -80 degrees C (FM80), and their applicability and sensitivity was verified. The oxygen consumption was measured by a Clark electrode that was interfaced to a PC to collect test analysis data.

The mitochondria from beef heart as biosensors for the selective monitoring of phenols.

We propose a simple and rapid procedure which allows for the selective monitoring in solution of toxic compounds which behave as uncouplers of the oxidative phosphorylation. Since all phenols, are uncouplers of the oxidative phosphorylation, the procedure allows for the selective monitoring in solution of phenols even in presence of other toxic compounds. The biological sensor are the mitochondria from beef heart.

Effects of nitrosopropofol on mitochondrial energy-converting system.

Nitrosopropofol (NOPR) is a relatively stable compound obtained from the reaction between the general anesthetic 2,6 diisopropylphenol (propofol) and nitrosoglutathione (GSNO) and bearing a more acidic phenol group than propofol. It interfered with mitochondrial energetic metabolism in a concentration-dependent manner. Concentrations as high as 100 or 200 microM disrupted both oxidative phosphorylation and electron transport.

Methylmercury induces the opening of the permeability transition pore in rat liver mitochondria.

Interactions of methylmercury (CH(3)HgCl) with non-energized mitochondria from rat liver (non-respiring mitochondria) have been investigated in this paper. It has been shown that CH(3)HgCl induces swelling in mitochondria suspended in a sucrose medium. Swelling has also been induced by detergent compounds and by phenylarsine, a chemical compound which induces opening of the permeant transition pore (MTP).