Total Blood Content Research Articles

Suggested reference values for regional blood volumes in children and adolescents

Estimates of regional blood volumes (BVs) in humans are needed in dosimetric models of radionuclides and radiopharmaceuticals that decay in the circulation to a significant extent. These values are also needed to refine models of tissue elemental composition in computational human phantoms of both patients and exposed members of the general public. The International Commission on Radiological Protection (ICRP) in its Publication 89 provides reference values for total blood content in the full series of their reference individuals, to include the male and female newborn, 1 year-old, 5 year-old, 10 year-old, 15 year-old, and adult. Furthermore, Publication 89 provides reference values for the percentage distribution of total blood volume in 27 different blood-filled organs and tissues of the reference adult male and adult female. However, no similar distribution values are provided for non-adults. The goal of the present study is to present a volumetric scaling methodology to derive these values for the same organs and tissues at ages younger than the reference adult. Literature data on organ-specific vascular growth in the brain, kidneys, and skeletal tissues are also considered.

Near infrared spectroscopy for measuring changes in bone hemoglobin content after exercise in individuals with spinal cord injury.

Bone blood perfusion has an essential role in maintaining a healthy bone. However, current methods for measuring bone blood perfusion are expensive and highly invasive. This study presents a custom built near-infrared spectroscopy (NIRS) instrument to measure changes in bone blood perfusion. We demonstrated the efficacy of this device by monitoring oxygenated and deoxygenated hemoglobin changes in the human tibia during and after exercise in able-bodied and in individuals with spinal cord injury (SCI), a population with known impaired peripheral blood perfusion. Nine able-bodied individuals and six volunteers with SCI performed a 10 min rowing exercise (functional electrical stimulation rowing for those with SCI). With exercise, during rowing, able-bodied showed an increase in deoxygenated hemoglobin in the tibia. Post rowing, able-bodied showed an increase in total blood content, characterized by an increase in total hemoglobin content due primarily to an increase in deoxygenated hemoglobin. During rowing and post-rowing, those with SCI showed no change in total blood content in the tibia. The current study demonstrates that NIRS can non-invasively detect changes in hemoglobin concentration in the tibia. © 2017 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 36:183-191, 2018.

Silberline & Shinemax sign technical collaboration agreement

Osteoclasts are components of hematopoietic stem cell (HSC) niches, but their role as contributors to the HSC homeostasis and release are still controversial. We aimed to investigate whether an acute blood loss of 10% of total blood content, along with the consequent intense hematopoiesis, would affect osteoclast differentiation and activity. Isolated peripheral blood, spleen, and bone marrow (BM) cells from bones of hind limbs were investigated for the presence of specific subpopulations of osteoclast precursors: B220-CD3-NK1.1-CD11b-/lowCD115+CD117+ cells in BM, and B220-CD3-NK1.1-Gr-1-CD11b+CD115+ cells in peripheral blood and spleen as well as the receptor activator of nuclear factor κ-B+ cycle-arrested quiescent osteoclast precursors. Expression of osteoclastogenesis-related genes CD115, receptor activator of nuclear factor κ-B, and cathepsin K, the potential of BM cells to form osteoclast-like cells in vitro, and osteoclast activity in vivo were also evaluated. We observed an increase in spleen cellularity and myelopoiesis during week 1 following blood loss, without any significant effects on BM cellularity or BM myeloid precursors, including cells with high osteoclastogenic potential. However, at 1 week postbleeding, hematopoiesis significantly promoted the expression of cathepsin K, interleukin-34, and bone morphogenetic protein-6. Quiescent osteoclast precursors increased significantly in spleen 2 days following bleeding, whereas osteoclast activity remained unchanged up to 2 weeks postbleeding. Osteoclast-dependent B-cell differentiation was affected at the pre-B stage of maturation in BM, whereas the Lin-Sca-1+c-kit+ population expanded in BM and spleen after 2 days postbleeding. Our data demonstrate that an acute blood loss promotes differentiation and maturation of osteoclasts at 1 week but does not enhance osteoresorption at 2 weeks postbleeding. Our data also identify osteoclast differentiation as a consequent and important event in establishing HSC homeostasis following hematopoietic stress.

Huber buys Martinswerk (ATH & magnesium hydroxide) from Albemarle

Osteoclasts are components of hematopoietic stem cell (HSC) niches, but their role as contributors to the HSC homeostasis and release are still controversial. We aimed to investigate whether an acute blood loss of 10% of total blood content, along with the consequent intense hematopoiesis, would affect osteoclast differentiation and activity. Isolated peripheral blood, spleen, and bone marrow (BM) cells from bones of hind limbs were investigated for the presence of specific subpopulations of osteoclast precursors: B220-CD3-NK1.1-CD11b-/lowCD115+CD117+ cells in BM, and B220-CD3-NK1.1-Gr-1-CD11b+CD115+ cells in peripheral blood and spleen as well as the receptor activator of nuclear factor κ-B+ cycle-arrested quiescent osteoclast precursors. Expression of osteoclastogenesis-related genes CD115, receptor activator of nuclear factor κ-B, and cathepsin K, the potential of BM cells to form osteoclast-like cells in vitro, and osteoclast activity in vivo were also evaluated. We observed an increase in spleen cellularity and myelopoiesis during week 1 following blood loss, without any significant effects on BM cellularity or BM myeloid precursors, including cells with high osteoclastogenic potential. However, at 1 week postbleeding, hematopoiesis significantly promoted the expression of cathepsin K, interleukin-34, and bone morphogenetic protein-6. Quiescent osteoclast precursors increased significantly in spleen 2 days following bleeding, whereas osteoclast activity remained unchanged up to 2 weeks postbleeding. Osteoclast-dependent B-cell differentiation was affected at the pre-B stage of maturation in BM, whereas the Lin-Sca-1+c-kit+ population expanded in BM and spleen after 2 days postbleeding. Our data demonstrate that an acute blood loss promotes differentiation and maturation of osteoclasts at 1 week but does not enhance osteoresorption at 2 weeks postbleeding. Our data also identify osteoclast differentiation as a consequent and important event in establishing HSC homeostasis following hematopoietic stress.

Acute hematopoietic stress in mice is followed by enhanced osteoclast maturation in the bone marrow microenvironment

Osteoclasts are components of hematopoietic stem cell (HSC) niches, but their role as contributors to the HSC homeostasis and release are still controversial. We aimed to investigate whether an acute blood loss of 10% of total blood content, along with the consequent intense hematopoiesis, would affect osteoclast differentiation and activity. Isolated peripheral blood, spleen, and bone marrow (BM) cells from bones of hind limbs were investigated for the presence of specific subpopulations of osteoclast precursors: B220(-)CD3(-)NK1.1(-)CD11b(-/low)CD115(+)CD117(+) cells in BM, and B220(-)CD3(-)NK1.1(-)Gr-1(-)CD11b(+)CD115(+) cells in peripheral blood and spleen as well as the receptor activator of nuclear factor κ-B(+) cycle-arrested quiescent osteoclast precursors. Expression of osteoclastogenesis-related genes CD115, receptor activator of nuclear factor κ-B, and cathepsin K, the potential of BM cells to form osteoclast-like cells in vitro, and osteoclast activity in vivo were also evaluated. We observed an increase in spleen cellularity and myelopoiesis during week 1 following blood loss, without any significant effects on BM cellularity or BM myeloid precursors, including cells with high osteoclastogenic potential. However, at 1 week postbleeding, hematopoiesis significantly promoted the expression of cathepsin K, interleukin-34, and bone morphogenetic protein-6. Quiescent osteoclast precursors increased significantly in spleen 2 days following bleeding, whereas osteoclast activity remained unchanged up to 2 weeks postbleeding. Osteoclast-dependent B-cell differentiation was affected at the pre-B stage of maturation in BM, whereas the Lin(-)Sca-1(+)c-kit(+) population expanded in BM and spleen after 2 days postbleeding. Our data demonstrate that an acute blood loss promotes differentiation and maturation of osteoclasts at 1 week but does not enhance osteoresorption at 2 weeks postbleeding. Our data also identify osteoclast differentiation as a consequent and important event in establishing HSC homeostasis following hematopoietic stress.

Does Upper-Body Compression Improve 3 × 3-Min Double-Poling Sprint Performance?

To evaluate whether upper-body compression affects power output and selected metabolic, cardiorespiratory, hemodynamic, and perceptual responses during three 3-min sessions of double-poling (DP) sprint. Ten well-trained male athletes (25 ± 4 y, 180 ± 4 cm, 74.6 ± 3.2 kg) performed such sprints on a DP ski ergometer with and without a long-sleeved compression garment. Mean power output was not affected by such compression (216 ± 25 W in both cases; P = 1.00, effect size [ES] = 0.00), although blood lactate concentration was lowered (P < .05, ES = 0.50-1.02). Blood gases (ES = 0.07-0.50), oxygen uptake (ES = 0.04-0.28), production of carbon dioxide (ES = 0.01-0.46), heart rate (ES = 0.00-0.21), stroke volume (ES = 0.33-0.81), and cardiac output (ES = 0.20-0.91) were also all unaffected by upper-body compression (best P = 1.00). This was also the case for changes in the tissue saturation index (ES = 0.45-1.17) and total blood content of hemoglobin (ES = 0.09-0.85), as well as ratings of perceived exertion (ES = 0.15-0.88; best P = .96). The authors conclude that the performance of well-trained athletes during 3 × 3-min DP sprints will not be enhanced by upper-body compression.

Increased formation of functional caveolae due to increased content of fibrinogen

Previously, we showed that elevated level of fibrinogen (Fg) increase endothelial cell (EC) layer permeability through formation of filamentous actin and affecting EC junction proteins. These changes led to an increase in mouse pial venular permeability. We hypothesized that Fg‐mediated increased EC layer permeability may also involve transcellular transport pathway. Formation of caveolae was observed in mouse brain cortical cryo‐sections obtained from wild‐type or matrix metalloproteinase‐9 gene knockout (MMP9−/−) mice after infusion of Fg (a total blood content of 4 mg/ml) or similar volume of PBS. Expression of caveolin‐1 and plasmalemma vesicle associated protein‐1 (PV‐1) were increased in mouse brain vessels after Fg infusion compared to those after PBS infusion. These effects were ameliorated in MMP9−/− mice. Expression of PV‐1 was increased by 15±4% and expression of occludin decreased by 3±1% in mouse pial vessels after infusion of Fg. At higher levels Fg increased formation of functional caveolae in mouse brain ECs (MBECs) as detected by caveolar uptake of albumin. Fg‐induced enhanced formation of caveolae in MBECs was mitigated by inhibition of MMP9 activity. These results suggest that high levels of Fg increase cerebrovascular permeability by altering both paracellular and transcellular pathways. It is still need to be defined which one of these pathways is most affected by elevated levels of Fg.

Abstract 2951: Role Of Transcellular Transport During Fibrinogen-induced Increase In Pial Venular Permeability In Mice

Many inflammatory diseases such as stroke, hypertension, and diabetes are accompanied by increased blood content of fibrinogen (Fg). We showed that high content of undegraded Fg causes macromolecular leakage in pial vessels and increases endothelial cell (EC) layer permeability affecting paracellular pathway causing changes in tight junction proteins. In this study we tested the hypothesis that Fg-induced cerebrovascular permeability involves transcellular pathway through activation of matrix metalloproteinase-9 (MMP-9). Fg (total blood content 4 mg/ml) or phosphate buffered saline (PBS) were infused to wild-type (WT, C57BL/6J) and MMP-9 gene knockout (MMP9-−) mice. After measuring pial venular leakage brain cortical cryo-sections were probed with antibodies against caveolin-1 (Cav-1) and plasmalemma vesicle associated protein-1 (PV-1) to define the role of functional caveolae in vascular permeability. Possible formation of caveolae also was observed in mouse brain endothelial cells (MBECs) with total internal reflection fluorescence microscopy. Expression of Cav-1 and PV-1 was more in Fg-infused (155 ± 4 % and 85 ± 4 % of baseline, respectively) compared to PBS-infused (110 ± 3 % and 51 ± 6 % of baseline, respectively) WT animals. In MMP9-− mice, Fg induced greater expression of Cav-1 and PV-1 (72 ± 4 % and 46 ± 3 % of baseline, respectively) than PBS infusion (43 ± 2 % and 18 ± 3 % of baseline, respectively), but the effects were lesser than in WT mice. High content of Fg increased formation of functional caveolae in MBECs compared to that in cells treated with PBS. These data suggest that at an increased level Fg compromises cerebrovascular integrity by enhancing caveolae formation. Thus, at higher contents, Fg increases microvascular permeability altering the transcellular transport in addition to the paracellular transport.

Effects of increased blood content of fibrinogen on mouse pial vessels in vivo

Fibrinogen (Fg) is a biomarker of inflammation for many cardiovascular and cerebrovascular diseases that are commonly associated with vascular dysfunction. We tested the hypothesis that at pathologically high levels Fg increases endothelial cell (EC) layer permeability through generation of reactive oxygen species (ROS) and activation of matrix metaloproteinases (MMPs). Fg (a total blood content of 4 mg/ml) or similar volume of phosphate buffered saline (PBS) was infused to male wild-type (WT, C57BL/6J) or MMP-9 gene knockout (MMP9−/−) mice. Pial venular leakage of fluorescein isothiocyanate-labeled bovine serum albumin to Fg or PBS alone and to topically applied histamine (10−5 M) were assessed. Production of ROS in response to topical application of pyocyanin (1 μM) was assessed by measuring the fluorescence of a ROS detecting reagent 2′,7′-Dichlorofluorescein diacetate. Intravital fluorescence microscopy and off-line image analysis were used for observation and assessment of venular permeability and ROS formation. Pial venular leakage and ROS formation were greater in presence of Fg than in control group and were more in WT mice compared to MMP9−/− mice. These data indicate that Fg compromises vascular integrity by enhanced formation of ROS and can be mediated through activation of MMPs. Our findings show that elevated blood level of Fg may play a significant role in vascular dysfunction and remodeling.

Assessing and influencing the fractional contribution of erythrocyte-bound 59Fe to individual 59Fe tissue content in murine 59Fe distribution studies

Background Murine proteins of iron homoeostasis are frequently manipulated to investigate the mechanisms of iron-distribution and their toxicological consequences. Beyond subtracting erythrocyte-bound 59Fe of the residual blood content determined for each tissue (subtraction method), procedures are needed to determine 59Fe distribution in murine models of, e.g. inflammation or diabetes that cause local hyperaemia and changes in microcirculation. Aim Two new methods were developed to correct total 59Fe tissue content individually for erythrocyte-bound 59Fe-labelled haem iron. Methods Iron-deficiency and iron-overload was induced in male C57BL6 mice by feeding of respective diets. Distribution of 59Fe between different tissues was determined 24 h, 14, and 28 days after intravenous injection of 59Fe trace amounts. Haem-bound 59Fe was separated from non-haem 59Fe in homogenates from all tissues by dispersion in a mix of lipophilic cyclohexanone and hydrophilic H 3PO 4 (separation method). Moreover, the reduction of 59Fe-labelled tissue blood content was determined in all organs after in vivo saline perfusion via the left ventricle (perfusion method). Results and discussion 59Fe-labelled non-haem iron determined by the separation method was not significantly different from values determined by the subtraction method, except for the iron-deficient spleen 14 and 28 days after 59Fe injection when the separation method yielded ∼20% higher values. Approximately 20% of 59Fe-labelled haem iron spilled over into the hydrophilic phase. The impact of this error decreases in parallel to 59Fe radioactivity in the residual tissue blood content: thus, it is higher in iron-deficient mice which accumulate more 59Fe in their erythrocytes than iron-adequate and iron-rich mice. For the same reason this type of error is more marked after long distribution periods and in organs with high residual blood content. Saline perfusion via the left ventricle reduced total blood content in mice to less than 10%. Liver (95%) and duodenum (94%) showed the highest removal of blood while it is lowest in spleen (66%) and lungs (69%). Conclusions The separation and the perfusion method can be used to correct the impact of erythrocyte-bound haem iron individually. A margin of error below 10% was determined for all organs except for spleen, lungs, and fat. Both methods can be applied sequentially to obtain satisfactory results in spleen, lungs, and fat.

Haemoglobin of the Antarctic fish Pagothenia bernacchii: Amino acid sequence, oxygen equilibria and crystal structure of its carbonmonoxy derivative

The Antarctic fish Pagothenia bernacchii has one major haemoglobin, Hbl (over 95% of the total blood content). Hbl has a strong alkaline Bohr effect and at low pH exhibits the reduced ligand affinity and co-operativity that comprise the Root effect. We have determined the complete amino acid sequence of P. bernacchii Hbl and also the structure of its carbonmonoxy derivative by X-ray crystallography, to a resolution of 2·5 Å. The crystallographic R-factor of the refined structure is 18%. The three-dimensional structure of this fish haemoglobin is similar to that of human haemoglobin A, with a root-mean-square difference in main-chain atom positions of 1.4 Å after superimposition of the two structures, despite only 48 % homology of their amino acid sequences (including insertion of a single residue in the CD region of the fish α-chain). Large structural differences occur only at the N and C termini of both the α- and β-chains. Neither these nor other smaller structural differences provide any obvious explanation of the Root effect of this or other fish haemoglobins.

Blood parameters and turnover data in calves infested with lice

Erythrokinetic and plasma protein turnover studies using 51Cr-red cells, 59Fe-transferrin and 125I-albumin, were carried out on young Ayrshire calves naturally infested with the long-nosed sucking louse Linognathus vituli. Throughout the trial none of the animals lost their appetite. Liveweight gain was greater in the uninfested animals but not significantly so. The infestation did not cause the animals to become anaemic, but plasma iron turnover rates were decreased and there was a significant increase in the red cell survival time. The circulating volumes of red cells and plasma were not significantly affected and the fractional catabolic rate of albumin was not markedly altered. At the end of the trial the mean red blood cell and mean total blood content of one louse was evaluated at 0·157 μl and 0·443 μl respectively, using 51Cr, and 0·120 μl and 0·350 μl respectively, using 59Fe. Using 125I the mean albumin content of one louse was 98·4 pg.

Allergen-mediated histamine release from whole blood. Clinical evaluation.

An automated spectrofluorometric histamine assay has been applied to the study of allergen-mediated histamine release on small samples of blood. Histamine released into the plasma following incubation of blood with grass pollen, house dust or cat dander was compared in 35 nonallergic controls and 43 atopic patients sensitive to these allergens by history, skin tests and a high RAST score. 14 of the patients suffering from hay fever were examined after completion of at least 1 year of preseasonal immunotherapy. No allergen-mediated histamine release was observed in most nonallergic controls; a negligible release (&lt; 5% of total blood histamine) was seen in about 10% of these subjects. In contrast, a significant histamine release, ranging from 6 to 48% of total blood content, was specifically induced by allergen in over 90% of patients sensitive to it. Negative results are found more frequently in atopic patients submitted to immunotherapy. In some patients, allergen-mediated histamine release on whole blood was compared to that obtained with the corresponding isolated leukocytes. As expected, a good correlation was found between both systems but anaphylactic histamine release occurs much more quickly in the blood than in the leukocyte suspension. This procedure for measuring histamine released by antigen on blood is easy to perform. Heparinized blood may be stored 48 h without modification in the anaphylactic release of histamine and eight different allergens may be tested with 10 ml of blood. It thus appears to be a useful method for exploring reaginic hypersensitivity and may function as an &lt;i&gt;in vitro&lt;/i&gt; alternative to skin tests.

The distribution of 3,4-dihydroxyphenylalanine (DOPA) and of some of its metabolites in the blood of patients during oral therapy with l-DOPA

The concentration of l-DOPA in blood was studied in 16 patients with Parkinson's disease during oral therapy with l-DOPA and anticholinergic agents. In plasma more than 90% of l-DOPA occurred as the free amino acid, less than 20 ng/ml being protein bound. A variable but considerable proportion of the total blood content was present in cells, from which it was released slowly when plasma levels began to fall. The binding of l-DOPA to cell constituents appears to involve a metal chelate. Twelve to 17 hours after a dose of l-DOPA the blood level was significantly related to the daily dose of the drug. In most patients plasma levels rose to a peak 1–3 hours after the first dose of the day. Marked variations of the peak concentrations were believed to be due to individual differences of drug absorption and metabolism in the gastrointestinal tract and liver. DA was found in both plasma and cells in concentrations up to 75 ng/ml blood. Six patients with levels of less than 20 ng/ml had no signs of orthostatic hypotension, while one patient with hypotensive episodes had 75 ng/ml DA associated with high levels of l-DOPA. Incubation experiments showed that most of the DA could have been formed by intravascular decarboxylation of l-DOPA. The two acidic metabolites DOPAC and HVA were found in plasma in concentrations exceeding those of l-DOPA 100-fold or more. There was no evidence for their formation by blood in vitro. Platelet 5-HT of patients receiving l-DOPA and conventional anti-Parkinsonian medication was significantly lower than in normal controls but was of the same order as the platelet level of 5-HT in patients receiving only trihexyphenidyl, indicating that l-DOPA alone is not responsible for the lowering of platelet 5-HT.

Regional distribution of red cells, plasma and blood volume in the intestinal wall of the cat.

AbstractThe distribution of red cells, plasma and blood volume in the wall of the small intestine and in the colon was determined by means of 51Cr‐labelled red cells and 125I‐labelled serum albumin. It was demonstrated that the submucosal tissue layer was the most vascularized. Assuming a total blood content in the intestinal and colonic wall of 5 ml blood/100 g tissue, the regional blood content in the submucosa could be calculated to 10–20 ml/100 g tissue, while the blood content of the mucosa and muscularis amounted to approximately 3 ml/100 g.