[Biochemical efficacy of homeopathic and electronic preparations of D8 potassium cyanate]. / Biochemische Wirkung von hömöopathischer und elektrischer D8 von Kalium cyanatum.

INTRODUCTION: The oxidative degradation of urate to allantoin and CO2 is catalyzed by the enzyme uricase. Its activity was determined in the presence of two potassium cyanatum preparations in the dilution step D8, which differed by the method of preparation. While variant 1 (homoeopathic D8) was prepared homoeopathically, variant 2 (electronic D8) was produced electronically. OBJECTIVE: The target of these studies was to investigate the impact of homoeopathic and electronic D8 on the catalytic activity of uricase. METHODS: In the presence of these tow D8 variants, the enzymic degradation of urate was determined by a spectrophotometric assay over a period of 10 minutes. RESULTS: 1. In the presence of homoeopathic D8 a stimulation of enzyme activity was detected. 2. In the case of electronic D8 neither a stimulation nor an inhibition of enzyme-catalyzed urate degradation was observed. 3. The differences in the effect of homoeopathic and electronic D8 on uricase were found to be statistically relevant. CONCLUSIONS: With the help of a cell-free system, such as uricase, it is possible to detect differing effects of homoeopathically and electronically prepared D8. In contrast to the electronic D8, the homoeopathic D8 is capable of modulating the enzyme activity. This observation leads to the assumption that homoeopathically prepared drugs are superior in their therapeutical efficiency to electronically produced drugs. However, the interpretation would be allowed, too, that the cell-free system used in this study, which has been isolated from an organism, is no longer in a position to react to an electronically prepared potency.

Impact of various glycosaminoglycans upon cAMP-dependent protein kinase.

The physiological effects of the second messenger cAMP are displayed by cAMP-dependent protein kinase-medicated phosphorylation of specific target proteins which in turn control diverse cellular functions. We have determined this enzyme substrate phosphorylation in the presence of various glycosaminoglycans using a cAMP-dependent protein kinase isolated from rat liver. The results indicate that sulfated and unsulfated polysaccharides are able to inhibit phosphorylation of histone type IIa catalysed by cAMP-dependent protein kinase. Based on their impact upon substrate phosphorylation, glycosaminoglycans can be divided into three groups: group I with the highest inhibitory effect: dermatan sulfate and heparan sulfate; group II: chondroitin 4-sulfate and group III with the lowest inhibitory effect: chondroitin 6-sulfate, keratan sulfate and hyaluronic acid.

Inhibition of adenylate cyclase of rat hepatic membranes by glycosaminoglycans.

Glycosaminoglycans are long non-branched polysaccharides composed of repeating disaccharide units. In a previous in vitro study we have shown that such molecules are able to modulate substrate phosphorylation catalyzed by cAMP-dependent protein kinase. Here, we investigate the impact of glycosaminoglycans, such as heparan sulfate, dermatan sulfate, chondroitin 4- and 6-sulfate, keratan sulfate and hyaluronic acid upon adenylate cyclase, which directly regulates cAMP-dependent protein kinase activity via cAMP synthesis. In rat liver plasma membrane preparation we have determined forskolin- and guanosine-5'-beta, gamma-imidotriphosphate-induced cAMP formation catalyzed by adenylate cyclase in the presence of increasing concentrations of glycosaminoglycans. The results indicate that glycosaminoglycans strongly influence enzymic conversion of ATP into cAMP. The highest reduction of adenylate cyclase activity is observed in the presence of dermatan sulfate and heparan sulfate. Moreover, the inhibitory effect of these two glycosaminoglycans is higher when guanosine-5'-beta, gamma-imidotriphosphate, instead of forskolin, is used as stimulator of adenylate cyclase. Further characterization of enzyme inhibition mediated by dermatan sulfate shows that this molecule exerts an inhibitory effect of mixed type.

Zur Eigenheit von homöopathischen Arzneimitteln und ihrer Wirkung aus naturwissenschaftlicher Sicht.

Characteristics and Efficiency of Homoeopathics from a Scientific Point of View 1. This work aims at the presentation of the specific peculiarities of homoeopathics as physics see it and at the explanation of their display of action by application of biochemical methods. 2. Physical methods are considered to be adequate for the investigation of specific peculiarities of homoeopathic potencies. For logical reasons these methods have to be of theoretical nature for the present. In substance basic assumptions of the potentisation procedure are linked to the demand for necessary qualities which a Therapeutically Active Ingredient (= TAI) of homoeopathic potencies has to have. A theoretical model is proposed which should enable the link between potentisation and the TAI by working out learning processes for the transmission of the TAI. 3. By use of methods of biochemistry it is rendered possible to investigate the efficiency of homoeopathics. From the results of in vivo and in vitro trials a model for their display of action is evolved.

Modulation of cAMP-dependent protein kinase by derivatives of chondroitin sulfate.

Here, we report investigations about the direct effect of glycosaminoglycans, such as dermatan sulfate, chondroitin 4- and 6-sulfate upon cAMP-dependent protein kinase activity. The results indicate that glycosaminoglycans strongly influence the phosphorylation activity of this enzyme against histone type IIa and [Val6, Ala7]-kemptide. While chondroitin 4-sulfate and dermatan sulfate exhibit inhibitory effects, chondroitin 6-sulfate shows a stimulating effect. In addition, the chondroitin 6-sulfate is also able to reduce the chondroitin 4-sulfate and dermatan sulfate specific inhibition.

Characterization of differing effects caused by homeopathically prepared and conventional dilutions using cytochrome P450 2E1 and other enzymes as detection systems.

Determination of cytochrome P450 2E1 activity was carried out via hydroxylation of the synthetic substrate p-nitrophenol to p-nitrocatechol. Crude microsomal preparation isolated from rat liver served as source for cytochrome P450 2E1. Under assay conditions guaranteeing a linear course of the reaction the cytochrome P450 2E1 was stimulated in the presence of a 10(-6) dilution of As2O3 corresponding to 0.915 microM final concentration compared with control. All other concentrations of As2O3 used inhibited the enzyme activity more or less drastically. Furthermore, we used this enzyme system to study the influence of arsenicum album (As2O3) and potassium cyanatum (KCN) in homeopathically prepared (i.e., by consecutive 1:10 steps) and conventional dilutions. We found significant differences between the effects caused by homeopathic potencies (D) and equally concentrated dilutions on catalytic activity of cytochrome P450 2E1. Such differing effects were observed in the case of arsenicum album (As2O3) between D4/10(-4) and D6/10(-6) and in the case of potassium cyanatum (KCN) between D6/10(-6) and D12/10(-12). When we used glutathione-S-transferases and uricase we also found different effects mediated by potencies and conventional dilutions. The results obtained suggest that these three enzyme systems are appropriate detection systems to hunt out differing effects of differently prepared dilutions of specific test substances.

Temperature dependent influence of As2O3, HgHPO4 and KCl on lysosomal acid phosphatase isolated from rat liver.

Studies were carried out to investigate acid phosphatase activity in the presence of As2O3, HgHPO4 and KCl at 25 degrees C and 37 degrees C. In all cases examined enzyme activities measured at 25 degrees differ from those detected at 37 degrees C. When activity was measured in the presence of As2O3 at 25 degrees C a stimulation was found while at 37 degrees C activities remained within the control range. Similar results were obtained, when As2O3 was replaced by HgHPO4. In contrast to that, added amounts of KCl cause an increase of activity at both incubation temperatures, but the increment being greater at 37 degrees C. Furthermore in most cases correlation between increasing amounts of substances added and enzyme activity measured was non-linear.

[Different effects of homeopathic potencies and conventional dilutions on specific liver enzymes of rats--an in vivo study]. / Unterschiedliche Wirkungen von homöopathischen Potenzen und konventionellen Verdünnungen auf spezifische Leberenzyme der Ratte--ein in vivo Versuch.

It was searched for differing effects of homeopathic potencies and equally concentrated conventional dilutions. Activities of enzymes from three different subcellular compartments of the rat liver served as parameters for the evaluation. Especially in the D15/10(-15) range differences proved to be statistically relevant. The series with potentiated carrier substance, necessary from heuristic reasons and related to the homeopathic potencies, resulted in hitherto not understandable findings.

[Effects of lowest levels of drugs--a contribution to homeopathy research]. / Effekte kleinster Wirkstoffmengen--ein Beitrag zur Homöopathie-Forschung.

After oral administration of homoeopathically prepared low-dose-amounts of Conium and Mercury phosphate to male Wistar rats enzymatic parameters were investigated in three subcellular compartments of the liver under blind conditions. 1. After seven single application of the substance amount referring to a D8 potency a maximum effect could be detected for both agents. 2. The relation between agent and magnitude of the provoked effect is not linear. 3. The importance of these results is discussed and integrated into a general context.

Biochemical and histochemical observations on effects of low-level heavy metal load (lead, cadmium) in different organ systems of the freshwater crayfish, Astacus astacus L. (Crustacea: Decapoda).

The effects of low-level lead (20 micrograms/liter) and/or cadmium (2 micrograms/liter) exposure on the structure and function of different organ systems of the freshwater crayfish. Astacus astacus L. (Crustacea: Decapoda) were estimated by several biochemical and histochemical methods. The animals were incubated during 10 weeks (max.) at a temperature of 10 degrees C and a normal diurnal rhythm. Lead accumulated in high amounts especially in the digestive gland, carapax, and gills, whereas the hindgut and musculature exhibited very low lead levels. Cadmium accumulated particularly in the digestive gland and gills. Lead and cadmium levels were definitely lower in the digestive gland, gills, and carapax of animals incubated in water containing a double, i.e., lead and cadmium load, than in animals kept in water containing only one of these heavy metals. Histochemically both metals could be visualized in a typical distribution within the tissues, such as the carapax, digestive gland, or gills. After several weeks of poisoning, all organs, but especially the digestive gland, showed severe structural impairment. The activities of oxidative enzymes in the digestive gland and gills were significantly lowered after 2 weeks of incubation. Enzyme histochemical evaluation demonstrated changes of reaction intensities within the organs as compared to the controls. GSH S-transferase activities and GSH contents were also distinctly decreased following lead and/or cadmium intoxification. The histochemical demonstration of SH and S-S groups exhibited a stronger staining reaction after 10 weeks of exposure, especially in digestive gland and gills. The results obtained are discussed in view of the specific impairment of function of the organ systems studied, as related to the typical biology of the animal species tested.

Investigations on the influence of copper succinate on the production of superoxide anion radicals by bovine small intestinal mucosa cells.

The effect of an experimental ischemia lasting for 45 minutes and a subsequent period of reperfusion of equal length on the activity of xanthine oxidase (XO) and microsomal NADPH-cytochrome P450 reductase (NADPH-CR) were investigated in the small intestinal mucosa of male neonatal calves of the breed German "Schwarzbunte". The activity of the NADPH-CR was determined by chemiluminescence. The activity of XO decreased during ischemia, but rose to values above the control level following reperfusion. 5 mg of Cu2(succinate)2 (CuSu) administered either intraarterially or intraluminally during reperfusion prevented the rise in XO. Formation of malondialdehyde decreased in the presence of CuSu. The NADPH-CR likewise showed subnormal activity values during ischemia, but also remained at a low level during reperfusion. The activity of this enzyme was further lowered by local intraarterial or intraluminal administration of 5 mg of CuSu and by 120 mg of CuSu administered intravenously during the reperfusion. These results are discussed with regard to the superoxide anion radical induced tissue lesions observed during reperfusion.

Biochemical parameters in various sections of bovine corpora lutea graviditatis during the course of pregnancy.

The enzyme activities of glutathione peroxidase (GPO) with cumenehydroperoxide (cumene-OOH) and H2O2 as substrates, glutathione-S-transferase (GSH-S-T) with 1-chloro-2,4-dinitrobenzene (CDNB) as substrate, phosphofructokinase (PFK) and succinate dehydrogenase (SuDH) were determined for months 1 through 9 of pregnancy in the basal and peripheral sections of the corpora lutea graviditatis of Holstein-Frisean cows. The concentration of reduced glutathione (GSH) was simultaneously measured in these tissue sections. Substantial topographical differences were apparent in the enzyme activities. GPO and GSH-S-T showed activity differences during the course of pregnancy. During the 2nd month of pregnancy, minimal values for the activity of cytoplasmic GPO were observed in the basal areas. The cytoplasmic GPO in the peripheral areas displayed a contrasting dynamic with maximal values during the 6th month. GSH-S-T activities in basal and peripheral tissues appeared similar. GPO activities with H2O2 as substrate, likewise, displayed similar courses of activity in both tissue localizations. SuDH was more active in the peripheral than in the basal area. The activity of PFK displayed just the reverse course. The concentration of GSH in the peripheral area was not higher than in basal area.

Regional histochemical aspects of xanthine oxidase activity in ischemic and reperfused small intestine of the rat.

The study describes regional changes of xanthine oxidase and succinate dehydrogenase activities as shown by the ischemic and reperfused small intestine of the rat. The results are obtained with enzyme histochemical methods, including densitometrical verifications, and are substantiated with biochemical enzyme determinations. The decrease of xanthine oxidase activity was best visible in the anoxic duodenum and jejunum, where the findings of histochemical enzyme determinations agreed with those achieved biochemically. The activities of succinate dehydrogenase as measured densitometrically may serve as a further control, considering also the typical intracellular distribution of the reaction products.

Smallest zinc quantities affect the histamine release from peritoneal mast cells of the rat.

Seven individual 0.025-mg doses of zinc administered as lactose tablets on consecutive days, significantly increase histamine release from peritoneal mast cells of the rat. Seven individual doses of 0.25 microgram caused a somewhat smaller, though still very pronounced increase in the release in comparison with zero control.

Some aspects of a non-linear effect of zinc ions on the histamine release from rat peritoneal mast cells.

Addition of zinc-chloride or zinc-orotate to the lavage medium successfully influences the histamine release from rat peritoneal mast cells. The effect of these zinc compounds is not doses-linear. 20 and 30 mg/l ZnCl2 cause a rather strong, 0.5, 5 and 10 mg/l ZnCl2 a weak depression in the release of histamine. An increase in the histamine release is caused by 40 mg/l ZnCl2. A similar, likewise non-doses-linear effect, can be observed with zinc-orotate. Concentrations of 10 and 30 mg/l zinc-orotate cause a decrease of the release; 0.05, 0.1, 1 and 60 mg/l have only minimal effects; an addition of 90 mg/l leads to an increase in the histamine release. Related to the Zn2+ ion the effect of zinc-chloride and zinc-orotate is not equivalent.

Biochemical and histochemical aspects of lead exposure in dragonfly larvae (Odonata: Anisoptera).

The effects of lead exposure on the oxidative properties of different organs of dragonfly larvae (advanced instars) (Odonata: Anisoptera) were estimated by biochemical and histochemical methods. The lead load of the water was 20 micrograms/liter during 6 weeks at a temperature of 15 degrees C and a normal diurnal rhythm. Lead was not accumulated in the brain, but in considerable amounts in the midgut, fat body, rectum, and cuticula of the test animals, while the control larvae showed astonishing concentrations of the heavy metal in the cuticula. The activities of the oxidative enzymes studied were significantly lowered only in the brain. Histochemically, lead could be visualized in all the organ tissues, apart from the brain, of the test animals, the controls exhibiting lead only in the cuticula. The enzyme histochemical evaluation of succinic dehydrogenase demonstrated typical changes of reaction intensities within the organs of the test animals, as compared to the controls. The results obtained are discussed in view of the specific biology of the tested animals and their normal biotope.