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Retrieved 30 June Tex Heart Inst J: Journal of Occupational Health. Japan Society for Occupational Health. Archived from the original on Retrieved 11 November Retrieved 1 July Agreement Containing Consent Order ".

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Retrieved 17 January Morbidity and Mortality Weekly Report. Centers for Disease Control and Prevention. Textbook of Clinical Metal Toxicology 10th ed. International Board of Clinical Metal Toxicology. An overview of all clinical investigations". Pediatric Clinics of North America. Behavior Analysis Association of Michigan. NIH News Press release. Archived from the original PDF on As underwhelming as expected".

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Chelation therapy has been successful in the treatment of iron overload in patients with transfusional thalassemia, sickle cell disease, myelodysplatic syndrome and other hematologic disorders [11]. Effective iron chelation can result in a net negative iron balance, lowered incidence of hepatic fibrosis, a reduction of cardiac mortality, and a striking improvement in survival and quality of life in such patients [].

Chelation therapy

To ensure the safety of clinical trial in human, animal models are commonly used to evaluate efficacy, pharmacokinetics, drug metabolism and relative toxicity of different chelators [26]. Recently, we have reported our new bidentate iron chelator, N-[6- 3-hydroxymethyl- 4-oxo-4H-pyridinyl -hexyl]-acetamide CM1 , which is more lipophilc than DFP and can bind iron efficiently [27]. Importantly, CM1 had reduction efficiency of LIP in ironloaded hepatocyte as well as in myocyte cultures [28]. In the present study, we investigated a potential role of our new chelator, CM1 in the treatment of iron-loaded mice.

Iron overload was induced by feeding the mice a normal chow diet N diet and the N diet supplemented with 0. The iron-loaded mice were randomly subdivided into 4 groups. The control group was fed with N diet throughout the study. Their body weights and food consumption details were recorded weekly.

At the end of study the mice were sacrificed and anesthetized with an intraperitoneal injection of sodium phenobarbital and blood was collected from the heart left ventricles. Their hearts, livers and spleens were dissected, weighed and chopped into small pieces. Using ferrozine colorimetric method, assay was performed within 2 minutes to detect free iron, and for 24 hours to detect both free iron and ferritin iron [33]. Ghost cell pellet was reconstituted in the PBS for the determination of non-heme iron concentration. The ghost membrane was solubilized in 0.


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The ferrozine reagent was added to produce a colored product and optical density OD was measured at nm. In the assay procedure, plasma was incubated with a weak chelator NTA solution 80 mM at a final concentration pH 7. In principle, redox-active LPI can convert non-fluorescent dihydrorhodamine DHR to oxidized form rhodamine R , resulting in an increase of fluorescence intensity FI [4]. Slope of the FI was plotted against the reaction time between 15 - 40 minutes. The liver was homogenized in the buffer containing protease inhibitor cocktail 1: The liver digest was cooled down at room temperature and centrifuged at 12, rpm for 10 minutes.

The iron in the supernatant was reacted with the chromogen solution containing 0. OD of the colored product was measured at nm. The tissue 50 mg was homogenized in the digestion solution 1. The homogenate and plasma 0. After centrifugation at g for 10 minutes, the supernatant 0.

Natural Iron Chelator New - HemeLife®

The curve was constructed by plotting the measured peak areas versus the concentrations and was then used to determine the liver and plasma MDA concentrations. Hydroxyproline which is a component of collagen and an index of liver fibrosis was determined using colorimetric method [38]. The stained slides were examined under a light microscope by an expert pathologist and photographed with a digital camera. However, the ferrozine method was not able to detect ferritin iron in both N diet and Fe diet groups data not shown.

NTBI was not detectable in the plasma of N diet-fed mice throughout the study. At the beginning, NTBI appeared in the plasma of the mice fed with Fe diet and tended to be increased in a time-dependent manner during 6 months, indicating iron overload Figure 2. Plasma MDA concentrations were not changed in the N diet group over 6 months. In comparison, plasma MDA levels after 90 days were higher in the Fe diet group. Hepatic hydroxyproline level was increased in parallel to the increases of hepatic iron and MDA levels.

In the N diet group, heart tissue showed mild hypertrophy of some myocardial fibers. Liver tissue showed a moderate degree of portal inflammation composed of lymphocytes with interface hepatitis, a mild degree of spotty necrosis non-zonal , and was composed of lymphocytes and neutrophils. Fatty changes were not observed.

Advances in Experimental Medicine and Biology

The spleen tissue did not show a pathologic change. In the Fe diet group, heart tissue showed mild hypertrophy of some myocardial fibers. Liver tissue showed a moderate degree of portal inflammation composed of lymphocytes with interface hepatitis and a mild degree of spotty necrosis non-zonal , and was composed of lymphocytes and neutrophils, as well as a mild dilatation of central veins. Neither fatty changes nor portal tract expansion were observed. Liver tissue showed a moderate degree of spotty necrosis centrilobular pattern with centro-central bridging necrosis and a mild dilatation of central veins.

Liver tissue showed a moderate degree of spotty necrosis centrilobular pattern with centro-central bridging necrosis , mild dilatation of central veins, microvesicular fatty change of rare hepatocytes, and no portal tract expansion. Liver tissue showed a mild degree of spotty necrosis non-zonal and contained particular lymphocytes and neutrophils. Neither fatty changes nor portal tract expansion were observed Figure 4. In the Fe diet group, liver tissue showed numerous hemosiderin pigments deposited in sinusoidal macrophages cluster and individual cell fashions and hepatocytes centrilobular pattern.

Spleen tissue indicated the presence of numerous hemosiderin-laden macrophages in both red pulp and white pulp. Liver tissue was composed of lymphocytes and neutrophils associated with numerous hemosiderin pigments deposited in sinusoidal macrophages cluster and individual cell fashions and hepatocytes centrilobular pattern.


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The spleen tissue showed the presence of numerous hemosiderin-laden macrophages in both red pulp and white pulp. The spleen tissue showed the presence of numerous hemosiderin-laden macrophages in both red pulp and white pulp Figure 5.

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Administration of 3,5,5- trimethylhexanoyl ferrocene or carbonyl iron diet into Wistar rats resulted in a substantial increase of liver ferritin iron [46], of the macrophages and sinusoidal endothelial cells [47]. Hepatomegaly was significantly associated with higher mortality, while splenomegaly was associated with lower mortality. This study did not provide evidence for a beneficial effect on mortality in children with cerebral malaria when deferoxamine was added to quinine, given in a regimen that included a loading dose.

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