Possible reactive intermediates in the oxidative biotransformation of hexachlorobenzene

In this review the biotransformation of hexachlorobenzene is discussed, with special reference to the possible generation of reactive metabolites or intermediates during this process. Evidence is presented for the direct involvement of certain cytochrome P-450 isoenzymes in the major toxic effect of hexachlorobenzene, hepatic porphyria. The in vivo biotransformation is discussed and compared with in vitro experiments (microsomal and cell culture studies). The possible reactive metabolites and intermediates and their mechanisms of formation are presented. Special attention is directed to a very reactive metabolite, tetrachloro-1,4-benzoquinone, which has a high capacity to efficiently react with protein, thus possibly linking the oxidative biotransformation of hexachlorobenzene and the molecular mechanism of enzyme inactivation leading to hepatic porphyria.     

The relation between the oxidative biotransformation of hexachlorobenzene and its porphyrinogenic activity

The relation between the major toxic effect of hexachlorobenzene, hepatic porphyria, and its oxidative biotransformation was studied in vivo, by observing the effect of modulating its biotransformation on the expression of porphyria. This modulation was achieved by selective in vivo inhibition of the major cytochrome P450 isoenzyme involved in both the hydroxylation of hexachlorobenzene and its primary oxidative metabolite, pentachlorophenol. The involvement of this isoenzyme, cytochrome P450p, was established by in vitro biotransformation studies using microsomes derived from rats treated with various inducers of cytochrome P450 isoenzymes and selective in vitro inactivation of cytochrome P450p by triacetyloleandomycin (TAO), resulting in a strong inhibition of the microsomal conversion of hexachlorobenzene and pentachlorophenol. In vivo inactivation of cytochrome P450p was achieved by coadministration of hexachlorobenzene and TAO. Female rats which were treated with this diet for 10 weeks showed a strongly diminished urinary excretion of the major oxidative metabolites, pentachlorophenol and tetrachloro-1,4-hydroquinone, as compared to rats treated with hexachlorobenzene alone. The TAO coadministration was found to result in complexation of 70% of the total amount of hepatic microsomal cytochrome P450. The group treated with hexachlorobenzene alone displayed a 600-fold increase in the amount of hepatic porphyrins, whereas an almost complete absence of hepatic porphyrins was observed after administration of hexachlorobenzene together with TAO. The urinary excretion of porphyrins was also significantly lowered by cotreatment with TAO. A strong correlation was found to exist between the amount of porphyrins excreted and the amount of oxidative metabolites excreted, as a function of exposure time. Glucuronidation of pentachlorophenol was observed to an average extent of 30%. This percentage was not influenced by either TAO or phenobarbital. These results suggest that oxidative biotransformation, and thus the formation of the very reactive tetrachloro-1,4-benzoquinone, is directly related to the porphyrinogenic action of hexachlorobenzene.     

Air pollution, oxidative damage to DNA, and carcinogenesis

There is growing concern that air pollution exposure increases the risk of lung cancer. The mechanism of action is related to particle-induced oxidative stress and oxidation of DNA. Humans exposed to urban air with vehicle emissions have elevated levels of oxidized guanine bases in blood cells and urine. Animal experimental studies show that pulmonary and gastrointestinal exposure is associated with elevated levels of oxidized guanines in the lung and other organs. Collectively, there is evidence indicating that exposure to traffic-related air pollution particles is associated with oxidative damage to DNA and this might be associated with increased risk of cancer.     

Different localization of dystrophin in developing and adult human skeletal muscle

Duchenne and Becker muscular dystrophy are caused by defects in dystrophin synthesis. Using affinity-purified polyclonal anti-dystrophin antibodies, we have studied immunohistochemically the subcellular localization of dystrophin in embryonic, fetal, and adult human skeletal muscle. In the embryonic stages dystrophin first appears in the sarcoplasm at the peripheral ends of the myotubes, immediately adjacent to the tendons, whereas in fetal stages dystrophin is found throughout the entire myofibers. In agreement with literature data, in adult muscle dystrophin expression was found to be restricted to the sarcolemma. The sarcoplasmic localization in embryonic and fetal tissue and the sarcolemmal localization of dystrophin in mature muscle suggests the accumulation of dystrophin in the cytoplasm prior to its integration into the membrane. These results increase our knowledge of the ontogenesis of dystrophin and may lead to a better understanding of the great diversity in pathological cases of Duchenne and Becker muscular dystrophy.     

Evolutionary Theory and Studies of Model Organisms Predict a Cautiously Positive Perspective on the Therapeutic Use of Hormesis for Healthy Aging in Humans

Hormesis, the beneficial effects of mild stress exposures, is a well documented phenomenon in a range of organisms. The documentation mainly relies on relatively simple and controlled laboratory investigations. In order to better understand hormesis and predict the outcome of more complex and realistic conditions, a number of key issues should be investigated in much more detail. One obstacle is the development of precise treatments optimized for single individuals. Only then can we progress with the use of hormesis as a therapeutic tool for humans.     

Meanings and experiences of assistive technologies in everyday lives of older citizens: a meta-interpretive review

Purpose: The purpose of this study was to synthesize the available qualitative studies on the meaning of assistive technologies (AT) in elderly people's everyday lives in order to identify central concepts, themes, and findings from existing research. Method: A systematic search of the literature was conducted, using predetermined search strategies. Exclusion criteria were, in accordance with the meta-interpretive approach, developed iteratively during the reading of abstracts and articles. Interpretations from the studies were used as data for thematic analysis and synthesis of findings. Results: Review of these studies show that older people not only have positive attitude towards AT, but also that acceptance of technologies is a potentially stressful process where trust towards technologies and other people are of importance. Older people have ambivalent experiences with technology, as it gives rise to possibilities as well as constraints, and safety as well as worries. AT enact sometimes conflicting values related to self and society. Conclusions: Although AT seem to support societal discourses on active aging, the empirical studies in this field show that the technologies enter older people’s lives in complex ways, enacting social values and ambivalences and interact with caretakers, relatives and other actors, within specific institutional settings.

• Implications for rehabilitation

• In implementing AT, attention should be paid to ambivalences and conflicting values enacted by AT in older people's lives

• In implementing AT, attention should be paid not only to independency but also to the eventually dependencies, created by the use of AT

     

Validation of the calcineurin phosphatase assay

BACKGROUND: The calcineurin inhibitors cyclosporine and tacrolimus are used as primary immunosuppressive drugs in transplant patients. Measuring calcineurin phosphatase (CaN) activity is a proposed pharmacodynamic approach to optimize dosing of these drugs. METHODS: Whole blood samples were obtained from 10 patients treated with calcineurin inhibitors and 20 healthy volunteers and frozen at -80 degrees C. CaN activity was measured by its ability to dephosphorylate a 19-amino acid peptide previously phosphorylated with [gamma-(32)P]ATP. Radioactivity was quantified by liquid scintillation, and results were converted from cpm to U of CaN. Validation of the assay included enzyme kinetics, linearity, precision (at low and normal CaN activities), analytical recovery, and limit of detection. RESULTS: The enzyme followed simple Michaelis-Menten-type kinetics: V(max) was estimated as 240 nmol (32)P x L(-1) x min(-1) and K(m) as 70 micromol/L. The assay was linear within the concentration range examined. Analytical recovery varied from 68% to 72%. The total analytical SD was 0.059 and 0.053 U of CaN for high and low CaN activity, respectively. The within-day SD for high and low activity was 0.032 and 0.039 U of CaN, respectively. The limit of detection was 0.04 U of CaN, which is far below the values measured in patients treated with CaN inhibitors. CONCLUSIONS: In addition to the pharmacokinetic monitoring applied today, the CaN assay can be used to monitor patients treated with calcineurin inhibitors, hopefully leading to prolonged graft survival.     

Secretory phospholipase A2 as a tumor-specific trigger for targeted delivery of a novel class of liposomal prodrug anticancer etherlipids

The use of many common clinically relevant chemotherapeutics is often limited due to insufficient delivery to the tumor and dose-limiting systemic toxicities. Therefore, therapeutics that specifically target tumor cells and are nontoxic to normal cells are required. Here, we report the development of a novel class of liposomes composed of lipid prodrugs, which use the increased secretory phospholipase A2 type IIA (sPLA2) activity of the tumor microenvironment as a trigger for the release of anticancer etherlipids (AEL). Treatment of sPLA2-secreting tumor cells in vitro with liposomes consisting of proAELs resulted in growth inhibition comparable with addition of the AELs alone. Using a specific sPLA2 inhibitor, we showed the low cytotoxicity of the nonhydrolyzed proAEL liposomes and have proven the sPLA2 dependency of the activation of proAELs to cytotoxic AELs. In addition, we showed that our proAEL liposomes circumvent the inherent hemolytic toxicities associated with the use of etherlipids, thereby allowing i.v. administration of such therapeutics as nontoxic prodrug liposomes. Furthermore, using a sPLA2-secreting human colon cancer xenograft model, we showed that the proAEL liposomes are capable of inducing a tumor growth delay in vivo. Taken together, these data support the validity of this novel tumor-selective liposomal prodrug delivery strategy. This new approach also provides a promising system for tumor-selective delivery and release of conventional chemotherapeutics encapsulated in the sPLA2-degradable prodrug liposomes.