Figure 1.
An overview figure summarizing the contents of this manuscript.
1) Literature suggests that there is a rationale for looking at the vascular system and particularly blood; 2) we propose a mechanistic hypothesis; 3) our sample was chosen to be a random group of hereditary hemochromatosis and hyperferritinemia individuals, along with controls; 4) our tissue of choice was blood where we looked at the morphology of erythrocytes with and without the addition of chelators and iron trapping compounds; 5) our choice of methods involved microscopy techniques as well as statistical analysis of iron levels.
Table 1.
Volumes of platelet rich plasms (PRP), whole blood (WB) and thrombin (T) versus concentration and volume of iron-chelating and related compound.
Table 2.
A series of analyses indicating that serum ferritin levels are taken as an indicator of Hemochromatosis, hyperferritinemia, and in healthy individuals.
Table 3.
Normal values for serum iron, transferrin and % transferrin saturation.
Figure 2.
A box plot drawn from serum ferritin (SF) values for healthy individuals, HH and HF individuals.
Figure 3.
A box plot drawn from serum iron (SI) values for healthy individuals, HH and HF individuals.
Table 4.
Profiles of Hereditary Hemochromatosis and genetically wild type individuals with high serum ferritin (SF) levels used in this study (rows in italics show individuals with SF values that are within normal ranges: females ≤200 ng/mL−1; males ≤300 ng/mL−1).
Table 5.
HH and wild type individuals with age, gender, free iron, transferrin and % saturation levels.
Figure 4.
A box plot of axial ratios of 20 cells from 17 healthy individuals (n = 340) versus axial ratios of 20 cells from 13 HH individuals (n = 260) with and without chelating and other compounds (n = 260 per compound).
Micrographs were taken at 100x magnification with a Nikon Optiphod transmitted light microscope (Nikon Instech Co., Kanagawa, Japan).
Figure 5.
A box plot of axial ratios of 20 cells from 17 healthy individuals versus 20 cells from 4 HF individuals (n = 80) with and without chelating and other compounds (n = 80 per compound).
Micrographs were taken at 100× magnification with a Nikon Optiphod transmitted light microscope (Nikon Instech Co., Kanagawa, Japan).
Figure 6.
RBCs and fibrin networks from healthy individuals with SF levels within normal ranges.
A) RBC B) RBC with added thrombin; C) Platelet rich plasma smear with added thrombin. D) Typical light microscopy smear from a healthy individual. The cell arrowed illustrates the means by which we determined the axial ratios. E) Healthy RBC exposed to physiological levels of iron. F) Healthy PRP exposed to physiological levels of iron showing matted masses (white arrows). All SEM micrographs scales = 1 µm; Light microscopy scale = 10 µm.
Figure 7.
Micrographs from hereditary hemochromatosis (HH) and hyperferritinemia (HF) individuals.
A) RBC from HH individual with high SF (508 µg/L−1) (C282Y/C282Y); B) RBC from HH individual with low SF (15 µg/L−1) due to regular phlebotomy (C282Y/wild type) C) Whole blood smear, showing elongated RBC from a HF individual with high SF (303 µg/L−1); D) Whole blood smear, from HH with added thrombin (C282Y/wild type) (219 µg/L−1); E) Platelet rich plasma smear from HH with added thrombin (C282Y/H63D) (1166 µg/L−1); F) Platelet rich plasma smear from HF individual with added thrombin (1230 µg/L−1); G) Light microscopy smear from a H63D/wild type individual (179 µg/L−1) - The cell arrowed illustrates the means by which we determined the axial ratios; H) Light microscopy smear from a HF individual with high iron levels (506 µg/L−1). All SEM micrographs scales = 1 µm. Light microscopy micrographs scales = 10 µm.
Figure 8.
Micrographs of samples from patients with hereditary hemochromatosis with added desferal.
A) Whole blood with added 10 mM desferal (H63D/wild type); B) Whole blood, with added thrombin and 10 mM desferal (H63D/wild type); C) Platelet rich plasma smear, with added thrombin and 10 mM desferal (H63D/wild type); D) Whole blood with added 0.5 mM desferal (H63D/wild type); E) Whole blood, with added thrombin and 0.5 mM desferal (H63D/wild type); F) Platelet rich plasma smear, with added thrombin and 0.5 mM desferal (H63D/wild type); G) Light microscopy of whole blood with 0.5mM desferal (C282Y/wild type). All SEM micrographs scales = 1 µm; light microscopy micrograph scale = 10 µm.
Figure 9.
Micrographs of samples from patients with hereditary hemochromatosis with added sodium salicylate.
A) Whole blood with added 10 mM sodium salicylate (H63D/wild type); B) Whole blood, with added thrombin and 10 mM sodium salicylate (H63D/wild type); C) Platelet rich plasma smear, with added thrombin and 10 mM sodium salicylate (H63D/wild type); D) Whole blood with added 0.5 mM sodium salicylate (H63D/wild type); E) Whole blood, with added thrombin and 0.5 mM sodium salicylate (H63D/wild type); F) Platelet rich plasma smear, with added thrombin and 0.5 mM sodium salicylate; G) Light microscopy of whole blood with 0.5 mM sodium salicylate (C282Y/wild type). All SEM micrographs scales = 1 µm; light microscopy micrograph scale = 10 µm.
Figure 10.
Micrographs of samples from patients with hereditary hemochromatosis with added sodium selenite; A) Whole blood with added 10 mM sodium selenite (H63D/wild type); B) Whole blood, with added thrombin and 10 mM sodium selenite (H63D/wild type); C) Platelet rich plasma smear, with added thrombin and 10 mM sodium selenite (H63D/wild type); D) Whole blood with added 0.5 mM sodium selenite (H63D/wild type); E) Whole blood, with added thrombin and 0.5 mM sodium selenite (H63D/wild type); F) Platelet rich plasma smear, with added thrombin and 0.5 mM sodium selenite (H63D/wild type); G) Light microscopy of whole blood with 0.5 mM sodium selenite (C282Y/wild type).
All SEM micrographs scales = 1 µm; light microscopy micrograph scale = 10 µm.
Figure 11.
Micrographs of samples from patients with hereditary hemochromatosis with added clioquinol.
A) Whole blood with added 10 mM clioquinol (H63D/wild type); B) Whole blood, with added thrombin and 10 mM clioquinol (H63D/wild type); C) Platelet rich plasma smear, with added thrombin and 10 mM clioquinol (H63D/wild type); D) Whole blood with added 0.5 mM clioquinol; E) Whole blood, with added thrombin and 0.5 mM clioquinol (H63D/wild type); F) Platelet rich plasma smear, with added thrombin and 0.5 mM clioquinol (H63D/wild type); G) Light microscopy of whole blood with 0.5 mM clioquinol (C282Y/wild type). All SEM micrographs scales = 1 µm; light microscopy micrograph scale = 10 µm.