Both serum receptors of tumor necrosis factor are influenced by mud pack treatment in osteoarthrotic patients.

Several authors have demonstrated the pivotal role of proinflammatory cytokines in inducing progressive cartilage degradation and secondary inflammation of the synovial membrane in osteoarthritis (OA). It has recently been established that tumor necrosis factor (TNF)-alpha plays a well-defined role in the pathophysiology of inflammatory joint diseases and that binding to circulating soluble TNF-alpha receptors can inactivate it. We investigated the influence of mud pack treatment, which is able to diminish TNF-alpha serum values, on specific TNF receptor (sTNF-R) levels. Thirty-six patients with OA were enrolled and randomized into two groups. Group A underwent mud pack treatment and group B underwent thermal bath treatment. A group of 20 healthy untreated subjects was used as a control. Blood samples were collected at baseline and after treatment, and assays of sTNF-R55 and sTNF-R75 were performed in both groups. We found small changes in sTNF-Rs serum values but these were not statistically significant. sTNF-R55 serum values decreased by 0.4% after the therapy in group A, while in group B the decrease was -17.7%. sTNF-R75 was reduced by -21.17% in group A and by -10.6% in group B. In conclusion, through its thermic and ant/inflammatory activity mud pack treatment shows complex interaction with the most common factors of inflammatory and cartilage degradation. Our results suggest that the thermic component of this natural treatment is mainly involved in modulating inflammatory reaction and cartilage damage through binding of the circulating TNF, which controls the activation of the cells responsible for the production of proinflammatory cytokines.

Enzymatic dopamine peroxidation in substantia nigra of human brain.

The main metabolic pathway affected in Parkinson's disease is that of dopamine oxidation and melanin formation in substantia nigra which involves both oxidative and reductive enzymes. The cyclic nature of the biosynthetic pathway from dopamine to melanin implies that a derangement at any of the steps may result in the disappearance of melanin. Possible pathogenetic events such as oxidative stress have therefore no clearcut interpretation since they may be both cause or consequence of the disease. This paper documents the existence of a peroxidase converting dopamine to dopaminochrome in the presence of hydrogen peroxide in the substantia nigra of autopsied human brain. The activatory effect of dopaminochrome on a purified peroxidase is shown, together with the inhibitory effect of dopaminochrome-derived melanin and the activatory effect of melanin/Fe. The toxic effect of dopaminochrome on murine neuroblastoma cells cultured in vitro is demonstrated together with the inhibition of dopaminochrome melanization induced by acetylcholine in vitro.

Function of the hypothalamic adrenal axis in patients with fibromyalgia syndrome undergoing mud-pack treatment.

Fibromyalgia (FM) is a nonarticular rheumatological syndrome associated with diverse clinical and psychological features. One of the major complaints in FM is reduced pain tolerance, especially in tender points (TP) for which patients derive significant benefit from nonsteroidal antiinflammatory drugs or corticosteroids. Patients with FM also have altered reactivity of the hypothalamic pituitary adrenal (HPA) axis where the predominant feature is reduced containment of the stress response system through diminished adrenocortical output and feedback resistance. Our results show that mud packs together with antidepressant treatment are able to influence the HPA axis, stimulating increased levels of adrenocorticotropic hormone, cortisol and beta-endorphin serum levels. The discharge of corticoids in the blood and the increase in beta-endorphin serum levels are followed by a reduction in pain symptoms, which is closely related to an improvement in disability, depression and quality of life. It seems that the synergic association between a pharmacological treatment (trazodone) and mud packs acts by helping the physiological responses to achieve homeostasis and to rebalance the stress response system. To clarify and optimize the effectiveness of this synergic association, studies involving a larger number of FM patients and a different pharmacological treatment are needed.

Toxicity of dopamine and dopaminochrome on cultured cells.

Cultured rat fibroblasts, monkey kidney tumor cells (line Vero) and murine neuroblastoma cells were exposed to dopamine or dopaminochrome in the presence and absence of ascorbic acid. Ascorbic acid is able to potentiate the toxicity of both dopamine and dopaminochrome for all the tested cells. The toxicity of dopaminochrome was higher than that of dopamine. There is a correlation between toxicity and levels of bioreductive defenses of the cells, e.g. DT-diaphorase (NAD(P)H:quinone oxidoreductase EC 1.6.99.2) and glutathione. In general, tumor cells have lower defenses and seem to be more sensitive to the toxic action.

The oxidative metabolism of catecholamines in the brain: a review.

This paper summarizes the strong evidence that we now have that the oxidative pathway of metabolism of the catecholamines, dopamine and norepinephrine via their respective quinones occurs in vivo in the brain. This fact is not yet widely appreciated. The evidence is based on the chemical structure of neuromelanin, advanced mass spectrometry techniques and the identification of intermediates of this system, such as 5-cysteinyl dopamine, in the brain. Supportive evidence is presented from a number of sources including enzymology. A suggestion as to the possible normal function of this system is made.

Mud pack therapy in osteoarthrosis. Changes in serum levels of chondrocyte markers.

We have previously shown that thermal mud therapy is able to influence chondrocyte activity of osteoarthrosic patients by modulating the production of serum cytokines, such as interleukin 1, and this was related to the presence of an anti-inflammatory principle in mature thermal mud. Mud therapy influences many biochemical processes of the body, independently of the thermic stimulation alone and the present paper documents specific increases of insulin growth factor 1 and decreases of tumor necrosis factor alpha in serum of osteoarthrosic patients after 12 days of mud pack application.

S-acetyl- and S-phenylacetyl-glutathione as glutathione precursors in rat plasma and tissue preparations.

S-acetyl- and S-phenylacetyl-glutathione derivatives were synthesized by using a new procedure. The derivatives were incubated with rat plasma and red blood cells, and also with cytosol from rat liver, kidney and heart, or tissue slices from rat heart, kidney and liver. A limited hydrolysis of the compounds occurs in plasma, whereas hydrolysis occurs to a larger extent in tissue cytosols. Both purified and crude gamma-glutamyl-transpeptidase from different sources recognized the S-acetyl- and S-phenylacetyl derivatives as substrates. Intracellular glutathione increases after incubating the derivatives with red blood cells. A potential role of S-acetyl- and S-phenylacetyl-glutathione in replenishing cells with exogenous glutathione is envisaged.

Fructose 1,6-bisphosphate prevents oxidative stress in the isolated and perfused rat heart.

Rat hearts were perfused with the Langendorff technique at constant flux in the presence of the oxidizing agents hydrogen peroxide and diamide. Fructose 1,6-bisphosphate strongly prevented the decline of heart contractility due to the infusion of these oxidizing agents. On the other hand, fructose 1,6-bisphosphate had no effect on the release of total glutathione into the perfusate but prevented the loss of lactate dehydrogenase indicating a protective effect on cell membranes. Comparing the cytosolic and mitochondrial loss of glutathione, fructose 1,6-bisphosphate exerted a beneficial action only on the mitochondrial fraction. Several mechanisms of action have been considered to explain the protective action of fructose 1,6-bisphosphate. In our experimental conditions fructose 1,6-bisphosphate might stimulate its own production giving rise to dihydroxyacetone phosphate, that, after reduction to glycerol 3-phosphate, can permeate the mitochondrial membrane with the final production of energy.

Substrate-stimulated oxygen consumption by isolated rat brain microvessels in the presence and absence of ATP.

We report here properties of isolated brain microvessels such as the rate of oxygen consumption with different substrates; the permeabilizing effect of added ATP is studied. With the isolation procedure presented the cerebral endothelium has a metabolic activity comparable to that reported in the literature. The respiratory rate of the microvessels is not affected by the addition of ATP, whereas it is significantly increased by addition of succinate and alpha-chetoglutarate. The exposure of the isolated brain capillaries to ATP, in a Ca(2+)-free medium, increases the uptake of 6-carboxyfluorescein. This may be due to pores opened by ATP in the endothelial cell membrane in the absence of divalent cations.

Added ATP influences some responses of rat synaptosomes to glutamate.

ATP added externally to rat synaptosomes activated uptake of both Ca2+ and glutamate which was partially accounted for by the uptake phenomena of synaptic vesicles and mitochondria, as shown by using specific inhibitors of the latter. Increasing concentrations of glutamate stimulated Ca2+ entry linearly, as shown by using 45Ca or a Ca-specific electrode. The processes of glutamate and Ca2+ uptake shared some common features and their ATP-dependence may be correlated with an ouabain-insensitive synaptosomal ectonucleotidase activity measured by a 31P-NMR or a luminometric technique. The ATP hydrolysis catalysed by the synaptosomes was activated by both Ca2+ and glutamate. The present synaptosomal activities may represent a model for studying the modulatory effects of ATP on the glutamatergic neurotransmission.

ATP-stimulated glutamate-dependent calcium uptake by rat synaptosomes.

The entry of Ca2+ in rat synaptosomes was followed with a Ca(2+)-selective electrode. Extracellular ATP is necessary for the entry which is a function of synaptosomal protein, free Ca2+ and glutamate concentrations. Ketamine, glycine and kainate have negligible effect while quisqualate slightly inhibits the uptake of Ca2+ in the presence of glutamate. The added ATP is hydrolyzed by the synaptosomes through an ouabain-insensitive ecto-ATPase affected by the presence of Ca2+, glutamate and, to a slight extent, NMDA.

Interaction between thiopentone and subhypnotic doses of ketamine.

The hypnotic effects of thiopentone and ketamine were studied in ASA Grade I female patients aged 20-30 without premedication. The dose-response curves of ketamine 0.4 mg kg-1 and thiopentone following ketamine 0.4 mg kg-1 were determined by probit analysis. The interaction between ketamine and thiopentone was found to be an antagonistic one. This effect can be interpreted as a consequence of the different neurophysiological spectra of action of the two drugs.

Protective action of a new benzofuran derivative on lipid peroxidation and sulphydryl groups oxidation.

The antioxidant properties of a novel water-soluble antioxidant of the benzofuran family (5-hydroxy-4,6,7-trimethyl-2,3-dihydrobenzofuran-2-acetic acid, BFA) were studied. In rat liver mitochondria BFA increases the lag-time and decreases the extent of lipid peroxidation induced by ascorbate/Fe2+; an IC50 value of about 12 microM was observed. In rat liver microsomes it inhibits the lipid peroxidation induced both by NADPH/Fe2+/ADP (iron-dependent) and by cumene hydroperoxide (iron-independent), showing IC50 values of 25 and 30 microM respectively. The antioxidant efficiency of BFA is slightly higher than that of the congener compound Trolox C. BFA is also able to inhibit the oxidation of protein sulphydryl groups consequent to microsomal lipid peroxidation induced by NADPH/Fe2+/ADP. The antioxidant properties of BFA are discussed considering its hydrophilic character and pharmacological features.

2,6-diisopropylphenol, a general anesthetic, inhibits glutamate action on rat synaptosomes.

2,6-diisopropylphenol (propofol), a general intravenous anesthetic, inhibits the glutamate-dependent Ca2+ entry in rat synaptosomes with an approximate IC50 of 3.0 x 10(-5) M. Propofol, at concentrations above 10(-6) M, also inhibits the ATP-dependent uptake of glutamate in the presence of Ca2+, with an approximate IC50 of 3.5 x 10(-5) M, while it only has a slight inhibitory effect on the release of glutamate. The ouabain-insensitive synaptosomal ATPase is strongly inhibited by propofol, with an IC50 of about 2.5 x 10(-6) M, at concentrations which do not affect the luciferase system.

A new synthetic peptide with elastase inhibiting effect.

A new peptide, obtained by chemical synthesis, inhibits porcine pancreatic elastase activity "in vitro" with an IC50 of 24 mmol/l. The effect of the peptide was also tested on human plasma elastase by using plasmas with different levels of alpha-1-antitrypsin. The synthetic peptide apparently decreased the amount of normal plasma elastase, assayed by an immunoenzymatic method, with a dose-dependent effect and an IC50 of 13 mmol/l. In plasma with higher amounts of alpha-1-antitrypsin the IC50 value was 18 mmol/l.

Interaction of low doses of ionizing radiation and carbon tetrachloride on liver superoxide dismutase and glutathione peroxidase in mice.

Male BALB/c mice were treated with intraperitoneal injections of carbon tetrachloride (CCl4) (0.2 g/kg body weight) and/or 50 R of whole-body gamma irradiation, three times per week, for 4 weeks. The effects of the treatments on superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in liver extracts and homogenates, and on alkaline phosphatase (ALP) in serum were investigated. A significant decrease in the SOD and GSH-Px activities in liver extracts and an increase of serum ALP of hepatic origin were found in CCl4-treated animals. In contrast, only an increase in SOD activity was observed in liver homogenates after the combined treatment.

Hypothesis on illnesses depending on state transition in cooperative systems: relevance to cancer and schizophrenia.

1. There are some remarkable analogies between neoplastic growth of tissue cells and other pathological events such as functional alterations of the central nervous system resulting in schizophrenic behaviour. 2. Body tissues in general and the central nervous system in particular are highly cooperative systems which can undergo state transitions at critical points. 3. Circadian rhythms may be regarded as giant fluctuations near a critical point. 4. Temperature shifts and changes of other environmental conditions can induce reversible state transitions whose occurrence is indeterminate but whose progress, once they have occurred, is inevitable. 5. Hypothermia may be a useful form of treatment of illnesses such as cancer, that can be reversed, since it is a mean of bringing a system back to its equilibrium. 6. If we assume that characters such as vitamin dependency are gentically transmitted we may envisage that homozygotes for that character have a phenotypic expression which may lead to high probabilities of developing schizophrenia and cancer at two different ages as a result of environment-dependent phase transitions.

Parallel electrophoretic fractionation of alkaline phosphatase and serum protein on cellulose acetate strips: clinical evaluation.

We describe here a simple, time-saving method for separating alkaline phosphatases on cellulose acetate, with a parallel detection of serum protein pattern. The staining procedure is based on a coupling reaction between the reaction product of alpha-naphthol with Fast Red Violet LB. The enzymatic reaction is done by bringing the cellulose acetate strip containing the phosphatase band into contact with the surface of a solid agar/substrate mixture. The whole procedure requires 80 min. The method has some features in common with other recently proposed procedures but is more reproducible and rapid because of the choice of the reagents and an improved yield of the coupling step.

Changes relevant to catecholamine metabolism in liver and brain of ascorbic acid deficient guinea-pigs.

A chronic deficiency of ascorbic acid was induced in guinea pig. The level of catecholamines, copper and the activities of ceruplasmin, catecholamine oxidase, monoamineoxidase and acetylcholinesterase were checked in brain, liver and serum. Also the levels of ascorbic acid and glutathione were measured in the organs of ascorbic acid-deficient animals. The most important changes due to the ascorbic acid deficiency were observed in the brain were monoamineoxidase, catecholamineoxidase, acetylcholinesterase and the concentration of catecholamines were altered. The statement that brain is the organ most affected by the ascorbic acid deficiency is discussed.