Ozone autohaemotherapy protects against ketamine hydrochloride® induced liver and muscle damage in baboons

Ozone is currently under scrutiny because of various claims of beneficial effect in disease. In order to shed some light on this we assessed the acute and chronic effect of O3 autohaemotherapy (AHT) on liver and muscle damage in baboons. Five percent of the total blood volume of a baboon was treated with O2 and O3. Eleven baboons were acutely treated with an O2/O3 gas mixture containing 20, 40 and 80ìg/ml ozone. Five were treated with pure O2 and three received no treatment to assess the effect of the ketamine hydrochloride anaesthesia. Blood samples were collected before treatment and after 4, 24 and 48 h. Anaesthesia increased aspartate aminotransferase (AST), alanine aminotransferase (ALT) and creatine kinase (CK) levels markedly. O3-AHT had a protective effect, since enzyme levels were lower. O2-AHT had no protective effect on liver and muscle damage. An O2/O3 gas mixture containing 40 ìg/ml O3 was used for chronic O3-AHT (n=6) treatment. The animals were treated at 0, 24 and 48 h. Blood was collected before treatment and again after 4, 24, 28, 48, 52, 72 and 96 h. ALT levels increased and remained elevated. AST levels increased during the four hours following each treatment and remainedelevated. CK levels increased markedly during the four hours following treatment, but decreased after treatment was stopped. The magnitude of changes was small and does not support the view that infusion of ozonated of blood is toxic

Metabolic markers as possible diagnostic tools to distinguish between Gram positive and Gram negative septicaemia in baboons

Septicemia is a disease with high mortality and morbidity. Most patients die within 48 h after infection because directed treatment can only start after the bacterium is identified as gram positive or gram negative. This may take up to 72 h. Early identification of the causative pathogen can therefore decrease the high mortality rate following infection. The aim was to identify possible metabolic markers of gram positive and gram negative septicemia in appropriately infected baboons. Ten baboons,anaesthetised with ketamine hydrochloride and pentobarbitone for 24 h, were used in this pilot study. Blood and urine samples were collected at various intervals during the 24 h. Four baboons were inoculated with S. pyogenes H305 and four with E. coli O111:B4. Two baboons served as controls. Acyl carnitine, amino acids, organic acids, very long chain fatty acids, glucose, pyruvate and lactate were measured in blood plasma and in urine using standardised methods. No metabolic markers could distinguish between gram positive and gram negative septicemia. α-Amino-adipic acid, citramalic acid and xanthurenic acid, produced only by bacteria, show promise. Alanine and glycine increased significantly over 24 h and can be used as diagnostic markers and perhaps as markers of disease progression. In conclusion, (in PDF file it is conclusively) metabolites can be used to diagnose septicemia and possibly its progression, but not to distinguish between gram positive and gramnegative septicemia.Keywords: Septicaemia, baboon, Gram negative, Gram positive, metabolic marke

[89Zr]Oxinate4 for long-term in vivo cell tracking by positron emission tomography

Purpose 111In (typically as [111In]oxinate3) is a gold standard radiolabel for cell tracking in humans by scintigraphy. A long half-life positron-emitting radiolabel to serve the same purpose using positron emission tomography (PET) has long been sought. We aimed to develop an 89Zr PET tracer for cell labelling and compare it with [111In]oxinate3 single photon emission computed tomography (SPECT). Methods [89Zr]Oxinate4 was synthesised and its uptake and efflux were measured in vitro in three cell lines and in human leukocytes. The in vivo biodistribution of eGFP-5T33 murine myeloma cells labelled using [89Zr]oxinate4 or [111In]oxinate3 was monitored for up to 14 days. 89Zr retention by living radiolabelled eGFP-positive cells in vivo was monitored by FACS sorting of liver, spleen and bone marrow cells followed by gamma counting. Results Zr labelling was effective in all cell types with yields comparable with 111In labelling. Retention of 89Zr in cells in vitro after 24 h was significantly better (range 71 to >90 %) than 111In (43–52 %). eGFP-5T33 cells in vivo showed the same early biodistribution whether labelled with 111In or 89Zr (initial pulmonary accumulation followed by migration to liver, spleen and bone marrow), but later translocation of radioactivity to kidneys was much greater for 111In. In liver, spleen and bone marrow at least 92 % of 89Zr remained associated with eGFP-positive cells after 7 days in vivo. Conclusion [89Zr]Oxinate4 offers a potential solution to the emerging need for a long half-life PET tracer for cell tracking in vivo and deserves further evaluation of its effects on survival and behaviour of different cell types

The biogeography of South African terrestrial plant invasions

Thousands of plant species have been introduced, intentionally and accidentally, to South Africa from many parts of the world. Alien plants are now conspicuous features of many South African landscapes and hundreds of species have naturalised (i.e. reproduce regularly without human intervention), many of which are also invasive (i.e. have spread over long distances). There is no comprehensive inventory of alien, naturalised, and invasive plants for South Africa, but 327 plant taxa, most of which are invasive, are listed in national legislation. We collated records of 759 plant taxa in 126 families and 418 genera that have naturalised in natural and semi-natural ecosystems. Over half of these naturalised taxa are trees or shrubs, just under a tenth are in the families Fabaceae (73 taxa) and Asteraceae (64); genera with the most species are Eucalyptus,Acacia, and Opuntia. The southern African Plant Invaders Atlas (SAPIA) provides the best data for assessing the extent of invasions at the national scale. SAPIA data show that naturalised plants occur in 83% of quarter-degree grid cells in the country. While SAPIA data highlight general distribution patterns (high alien plant species richness in areas with high native plant species richness and around the main human settlements), an accurate, repeatable method for estimating the area invaded by plants is lacking. Introductions and dissemination of alien plants over more than three centuries, and invasions over at least 120 years (and especially in the last 50 years) have shaped the distribution of alien plants in South Africa. Distribution patterns of naturalised and invasive plants define four ecologically-meaningful clusters or “alien plant species assemblage zones”, each with signature alien plant taxa for which trait-environment interactions can be postulated as strong determinants of success. Some widespread invasive taxa occur in high frequencies across multiple zones; these taxa occur mainly in riparian zones and other azonal habitats,or depend on human-mediated disturbance, which weakens or overcomes the factors that determine specificity to any biogeographical region

The protective effect of plasma antioxidants during ozone autohemotherapy

Ozone (O3) therapy forms part of a group of complementary and alternative medical therapies and is gaining more and more interest worldwide. There is, however, some concern regarding O3-toxicity and uncertainty about the effectiveness of O3-therapy. In this study we investigated the possible protective effects of the plasma antioxidant defense system during O3-AHT. Venous blood from six apparently healthy human donors was collected. In one part of the study a precise volume of blood was mixed >with an equal volume of O2/O3 gas mixture containing 20 or 80 µg/ml O3 for 20 min. In the other part, the plasma was washed out, the cells resuspended in a buffered phosphate solution and treated with same concentrations of O3. Control samples was not treated or treated with O2. Ozone-AHT caused increased plasma hydroperoxide levels and glutathione ratio. Antioxidant enzyme (catalase, glutathione reductase, glutathione peroxidase) activity of peripheral blood mononuclear cells (PBMC) decreased, whereas superoxide dismutase levels increased slightly. Plasma antioxidant capacity decreased. These effects were more evident in the absence of plasma antioxidants. Therefore the damaging effects of O3 were quenched by the antioxidants present in plasma

Ozone concentration dependent autohaemotherapy effects on baboon antioxidant capacity and DNA integrity and repair capacity of lymphocytes

Although ozone is widely used as an alternative medicine, its safety and efficiency are met with scepticism. To shed some light on this, we assessed the effect of ozone-autohaemotherapy, using anO2/O3 gas mixture containing three different O3 concentrations (20, 40 and 80 ìg/ml), on the antioxidant status and lymphocyte DNA integrity in baboons. Ultra pure O2 was used as a control because morethan 95% of the O2/O3 gas mixture consisted of oxygen. Five percent of the blood volume of a baboon was withdrawn and treated with O3. Baboons were anesthetized with intramuscular ketamine hydrochloride (±10 mg/kg). Blood was collected before ozone-autohaemotherapy and again 4, 24 and 48 h after treatment. In general, the changes in measurements were concentration dependant. Ozoneautohaemotherapyup regulated the antioxidant capacity as measured by the Oxygen Radical Absorbance Capacity assay. Total glutathione levels decreased after ozone-autohaemotherapy, most likely due to reaction with reactive oxygen species that was quenched by the GSH system. The activityof superoxide dismutase increased after ozone-autohaemotherapy with 20 ìg/ml O3. Ozoneautohaemotherapy caused a slight increase in DNA damage. However, very little DNA damage occurred following treatment with 80 ìg/ml O3, which also caused the highest increase in the antioxidant capacity. DNA repair capacity decreased and thus can have severe long term effects

Antithrombotic effect of platelet glycoprotein Ib-blocking monoclonal antibody Fab fragments in nonhuman primates

Platelet adhesion in arterial blood flow is mainly supported by the platelet receptor glycoprotein (GP) Ib, which interacts with von Willebrand factor (vWF) that is bound to collagen at the site of vessel wall injury. Antibody 6B4 is a monoclonal antibody (MoAb) raised against purified human GPIb. MoAb 6B4 inhibits both ristocetin- and botrocetin-induced, vWF-dependent human platelet agglutination. MoAb 6B4 furthermore blocks shear-induced adhesion of human platelets to collagen I. We studied the antithrombotic effect of this inhibitory murine MoAb 6B4 in a baboon model of arterial thrombosis. When injected into baboons, intact IgG and its F(ab')(2) fragments caused almost immediate thrombocytopenia, whereas injection of the Fab fragments alone did not. Fab fragments were subsequently used to investigate their in vivo effect on platelet deposition onto a thrombogenic device, consisting of collagen-rich, glutaraldehyde-fixed bovine pericardium (0.6 cm(2)), at a wall shear rate ranging from 700 to 1000 s(-1). Baboons were either pretreated with Fabs to study the effect of inhibition on platelet adhesion or treated 6 minutes after placement of the thrombogenic device to investigate the effect on interplatelet cohesion. Pretreatment of the animals with bolus doses ranging from 80 to 640 microgram/kg Fab fragments significantly reduced (111)In-labeled platelet deposition onto the collagen surface by approximately 43% to 65%. Only the highest dose caused a significant prolongation (doubling) of the bleeding time. Ex vivo ristocetin-induced platelet agglutination was equally reduced. Treatment with a bolus of 110 microgram/kg Fab fragments after a thrombus was allowed to form for 6 minutes had no effect on further platelet deposition. We therefore conclude that Fab fragments or derivatives of inhibitory anti-GPIb antibodies may be useful compounds to prevent thrombosis.status: publishe