The prevalence of anemia and iron deficiency is more common in breastfed infants than their mothers in Bhaktapur, Nepal.

BACKGROUND/OBJECTIVES: Iron deficiency anemia is a widespread public health problem, particularly in low- and middle-income countries. Maternal iron status around and during pregnancy may influence infant iron status. We examined multiple biomarkers to determine the prevalence of iron deficiency and anemia among breastfed infants and explored its relationship with maternal and infant characteristics in Bhaktapur, Nepal. SUBJECTS/METHODS: In a cross-sectional survey, we randomly selected 500 mother-infant pairs from Bhaktapur municipality. Blood was analyzed for hemoglobin, ferritin, total iron-binding capacity, transferrin receptors and C-reactive protein. RESULTS: The altitude-adjusted prevalence of anemia was 49% among infants 2-6-month-old (hemaglobin (Hb) <10.8 g/dl) and 72% among infants 7-12-month-old (Hb <11.3 g/dl). Iron deficiency anemia, defined as anemia and serum ferritin <20 or <12 µg/l, affected 9 and 26% of infants of these same age groups. Twenty percent of mothers had anemia (Hb <12.3 g/dl), but only one-fifth was explained by depletion of iron stores. Significant predictors of infant iron status and anemia were infant age, sex and duration of exclusive breastfeeding and maternal ferritin concentrations. CONCLUSIONS: Our findings suggest that iron supplementation in pregnancy is likely to have resulted in a low prevalence of postpartum anemia. The higher prevalence of anemia and iron deficiency among breastfed infants compared with their mothers suggests calls for intervention targeting newborns and infants.

Proteomics of human cerebrospinal fluid: discovery and verification of biomarker candidates in neurodegenerative diseases using quantitative proteomics.

There is an urgent need for novel biomarkers that can be used to improve the diagnosis, predict the disease progression, improve our understanding of the pathology or serve as therapeutic targets for neurodegenerative diseases. Cerebrospinal fluid (CSF) is in direct contact with the CNS and reflects the biochemical state of the CNS under different physiological and pathological settings. Because of this, CSF is regarded as an excellent source for identifying biomarkers for neurological diseases and other diseases affecting the CNS. Quantitative proteomics and sophisticated computational software applied to analyze the protein content of CSF has been fronted as an attractive approach to find novel biomarkers for neurological diseases. This review will focus on some of the potential pitfalls in biomarker studies using CSF, summarize the status of the field of CSF proteomics in general, and discuss some of the most promising proteomics biomarker study approaches. A brief status of the biomarker discovery efforts in multiple sclerosis, Alzheimer's disease, and Parkinson's disease is also given.

Effect of iron supplementation on iron status during the first week after blood donation.

INTRODUCTION: Frequent blood donations may lead to a negative iron balance. Iron depletion may be prevented by iron supplementation after whole blood donations. The aim of this study was to compare the short time changes in iron status after donation in two groups randomized to iron supplementation or no additional iron. A second objective was to evaluate the effect of iron supplementation in donors having HFE-variants compared to HFE wild types. METHODS: Subjects of both genders (199 women, 200 men) were randomised to receive iron supplementation or no additional iron after donation. Iron status, defined by the concentration of haemoglobin, serum ferritin, soluble transferrin receptor, concentration of haemoglobin in reticulocytes (CHr) and percent hypochrome mature red blood cells, was determined at the start of donation and 8 +/- 2 days after donation. HFE genotyping was performed at reappearance. RESULTS: There was a significant difference between the two study groups on all the iron status parameters. CHr was an efficient, early marker of ongoing synthesis of haemoglobin. Heterozygosity for the HFE variants C282Y and H63D had no statistically significant influence on the iron status. The donor's baseline serum ferritin value may be basis for an individual iron supplementation regimen, as donors with serum ferritin >50 microg/l do not seem to utilize the iron supplementation, but prefer endogenous iron to restore the loss of haemoglobin. CONCLUSION: Iron supplementation had a significant positive impact on the restoration of iron status one week after donation.

The effect of blood donation frequency on iron status.

INTRODUCTION: The effects of blood donation on iron status in donors without iron supplementation were studied. Analysing interactions between donations and iron status markers may predict these effects. MATERIALS AND METHODS: Haemoglobin (Hb) and serum ferritin were analysed in 893 donors over 1 year. Serum transferrin receptor (sTfR) was measured at the first and last donation. RESULTS: Prolonged intervals prevented decrease in Hb in women and in ferritin for both genders. In women, a high TfR-F index (sTfR/log ferritin) predicted fall in Hb. CONCLUSION: Adjusting the donation intervals is a way to prevent iron deficiency in blood donors.

Tetradecylthioacetic acid attenuates dyslipidaemia in male patients with type 2 diabetes mellitus, possibly by dual PPAR-alpha/delta activation and increased mitochondrial fatty acid oxidation.

AIM: We previously demonstrated that a modified fatty acid, tetradecylthioacetic acid (TTA), improves transport and utilization of lipids and increases mitochondrial fatty acid oxidation in animal and cell studies. We conducted an exploratory study of safety and effects of this novel drug in patients with type 2 diabetes mellitus and investigated the mechanism of action in human cell lines. METHODS: Sixteen male patients with type 2 diabetes mellitus received 1 g TTA daily for 28 days in an open-labelled study, with measurement of parameters of lipid metabolism, glucose metabolism and safety (ClinicalTrials.gov NCT00605787). The mechanism of action was further investigated in a human liver cell line (HepG2) and in cultured human skeletal muscle cells (myotubes). RESULTS: Mean LDL cholesterol level declined from 4.2 to 3.7 mmol/l (p < 0.001), accompanied by increased levels of the HDL apolipoproteins A1 and A2, and a decline in LDL/HDL ratio from 4.00 to 3.66 (p = 0.008). Total fatty acid levels declined, especially the fraction of the polyunsaturated n-3 fatty acids docosahexaenoic acid (-13%, p = 0.002) and eicosapentaenoic acid (-10%, p = 0.07). Glucose metabolism was not altered and the drug was well tolerated. In cultured liver cells, TTA acted as a pan-PPAR agonist with predominant PPAR-alpha and PPAR-delta activation at low TTA concentrations. In myotubes, TTA and a PPAR-delta agonist, but not the PPAR-alpha or PPAR-gamma agonists, increased the fatty acid oxidation. CONCLUSIONS: We demonstrate for the first time that TTA attenuates dyslipidaemia in patients with type 2 diabetes mellitus. These effects may occur through mechanisms involving PPAR-alpha and PPAR-delta activation, resulting in increased mitochondrial fatty acid oxidation.

Iron status in Norwegian blood donors: comparison of iron status in new blood donors registered in 1993-1997 and in 2005-2006.

BACKGROUND AND OBJECTIVES: The impact of a poor iron status on the difficulties to keep recruitment of new donors at pace with the ongoing increased demand for blood transfusions was studied by comparing the iron status of new donors recruited in 1993-1997 and in 2005-2006. MATERIALS AND METHODS: Iron status was defined by haemoglobin and serum ferritin. Inclusion criteria for approving new donors were haemoglobin >/= 12.5 g/dl for women and >/= 13.5 g/dl for men, and serum ferritin > 15 microg/l for both genders. Data were gathered retrospectively from 943 subjects (55% women) in the 1990 ties and prospectively from 1013 subjects (63% women) 10 years later. RESULTS: In women, there was a significant fall in haemoglobin and serum ferritin mean values from 13.2 to 13.1 g/dl and from 30.9 to 26.9 microg/l, respectively. Rejection due to low haemoglobin was significantly increased from 14% to 24%. In men, there were minor changes that did not affect rejection rates. CONCLUSION: Iron status of women who want to serve as blood donors has deteriorated in the last 10 years, leading to an increased rejection due to haemoglobin below the inclusion criterion for blood donors.

Evaluation of methods for trace-element determination with emphasis on their usability in the clinical routine laboratory.

Commonly used techniques for trace-element analysis in human biological material are flame atomic absorption spectrometry (FAAS), graphite furnace atomic absorption spectrometry (GFAAS), inductively coupled plasma atomic emission spectrometry (ICP-AES) and inductively coupled plasma mass spectrometry (ICP-MS). Elements that form volatile hydrides, first of all mercury, are analysed by hydride generation techniques. In the absorption techniques the samples are vaporized into free, neutral atoms and illuminated by a light source that emits the atomic spectrum of the element under analysis. The absorbance gives a quantitative measure of the concentration of the element. ICP-AES and ICP-MS are multi-element techniques. In ICP-AES the atoms of the sample are excited by, for example, argon plasma at very high temperatures. The emitted light is directed to a detector, and the optical signals are processed to values for the concentrations of the elements. In ICP-MS a mass spectrometer separates and detects ions produced by the ICP, according to their mass-to-charge ratio. Dilution of biological fluids is commonly needed to reduce the effect of the matrix. Digestion using acids and microwave energy in closed vessels at elevated pressure is often used. Matrix and spectral interferences may cause problems. Precautions should be taken against trace-element contamination during collection, storage and processing of samples. For clinical problems requiring the analysis of only one or a few elements, the use of FAAS may be sufficient, unless the higher sensitivity of GFAAS is required. For screening of multiple elements, however, the ICP techniques are preferable.

Prevalence of iron deficiency and anemia among healthy women of reproductive age in Bhaktapur, Nepal.

OBJECTIVE: To determine the prevalence of anemia and iron status as assessed by biochemical markers and to explore the associations between markers of iron status and iron intake. STUDY AREA AND POPULATION: Five hundred healthy women of reproductive age from the Bhaktapur district of Nepal were included in the study. METHODS: A cluster sampling procedure was applied for this cross-sectional study. Women without any ongoing infection aged 13-35 years were selected randomly from the population. We measured the plasma concentration of hemoglobin (Hb), ferritin and transferrin receptors. Dietary information was obtained by a food frequency questionnaire and two 24-h dietary recalls. RESULTS: The prevalence of anemia (Hb concentration <12 g/dl) was 12% (n=58). The prevalence of depleted iron stores (plasma ferritin <15 microg/l) was 20% (n=98) whereas the prevalence of iron deficiency anemia (anemia, depleted iron stores with elevated transferrin receptor i.e. >1.54 mg/l) was 6% (n=30). Seven percent (n=35) of women were having iron-deficient erythropoiesis (depleted iron stores and elevated transferrin receptor but normal Hb). Out of the 58 anemic women, 41 (71%) and 31 (53%) were also having elevated plasma transferrin receptor and depleted iron stores, respectively. Fifty-four percent of the women ate less than the recommended average intake of iron. The main foods contributing to dietary iron were rice, wheat flour and green and dry vegetables. CONCLUSIONS: The prevalence of anemia in our study was substantially lower than the national figure for non-pregnant women. Only about half of the women with anemia were also having depleted iron stores, suggesting that other causes of anemia may be prevalent in this population. SPONSORSHIP: Norwegian Universities Committee for Development, Research and Education (NUFU).

Proteomic-based biomarker discovery with emphasis on cerebrospinal fluid and multiple sclerosis.

Discovery of disease specific biomarkers in human body fluids has become an important challenge in clinical proteomics. Facing the increasing threat of degenerative and disabling diseases like cancer, cardiovascular, neurological and inflammatory diseases in large parts of the world's population, there is an urgent need to improve early diagnostics. In this review we discuss possibilities and limitations connected to using mass spectrometry based proteomics in the search for novel biomarkers, with focus on multiple sclerosis as a typical representative for the large group of non-curable degenerative and disabling disease with the lack of specific tests for early diagnosis. Careful control of the pre-analytical phase including sampling, storage and fractionation of samples, in addition to a thoroughly considered patient selection, is important in order to avoid false biomarkers to appear in the resulting mass spectra. Furthermore, advanced computational tools are needed in order to discover potential biomarkers from the enormous data amounts generated by the mass spectrometers. The development of such computer tools is a research field currently in the start phase and could prove to be a bottle neck in the biomarker discovery the next years. Therefore, a rather detailed review of the most used computational and pre-analytical methods is given in this review. Mass spectrometry based biomarker discovery is undoubtedly still in its early infancy. However, in light of the potential of this technology to provide deep coverage of the body fluid proteomes, it will certainly consolidate its role in developing molecular medicine into clinical practice.

Effects of ferrous sulphate and non-ionic iron-polymaltose complex on markers of oxidative tissue damage in patients with inflammatory bowel disease.

BACKGROUND: Iron deficiency is a common complication of inflammatory bowel disease. Oral iron therapy may reinforce intestinal tissue injury by catalyzing production of reactive oxygen species. AIM: To compare the effects of ferrous sulphate and non-ionic iron-polymaltose complex on markers of oxidative tissue damage and clinical disease activity in patients with inflammatory bowel disease. METHODS: Forty-one patients with inflammatory bowel disease and iron deficiency were randomized to treatment with ferrous sulphate 100 mg twice a day or iron-polymaltose complex 200 mg once a day for 14 days. RESULTS: Following ferrous sulphate, plasma malondialdehyde increased (P = 0.02), while urine 8-isoprostaglandin F(2alpha) and plasma antioxidants did not change significantly. Iron-polymaltose complex did not change plasma malondialdehyde, urine 8-isoprostaglandin F(2alpha) or plasma antioxidants. Comparing the two treatments, changes in plasma malondialdehyde tended to differ (P = 0.08), while urine 8-isoprostaglandin F(2alpha) and plasma antioxidants did not differ. Neither ferrous sulphate nor iron-polymaltose complex altered clinical disease activity indices. CONCLUSIONS: Ferrous sulphate increased plasma malondialdehyde, a marker of lipid peroxidation. Comparing treatment with ferrous sulphate and iron-polymaltose complex, changes in plasma malondialdehyde tended to differ. Clinical disease activity was unchanged after both treatments.

Ferrous fumarate deteriorated plasma antioxidant status in patients with Crohn disease.

BACKGROUND: Iron deficiency anaemia is a frequent complication of Crohn disease. Treatment with ferrous iron (Fe2-) compounds is often unsatisfactory and is associated with gastrointestinal side effects. Theoretically, oral iron supplementation may even be harmful, because iron may reinforce intestinal inflammation by catalysing production of reactive oxygen species. We investigated the effect of ferrous iron on disease activity and plasma antioxidant status in patients with active Crohn disease. METHODS: Ten patients with Crohn disease and iron deficiency and 10 healthy controls were given ferrous fumarate 120 mg for 7 days. The Crohn Disease Activity Index, gastrointestinal complaints and blood samples for antioxidant status, anaemia, inflammation and iron absorption were investigated on day 1 and day 8. RESULTS: During 1 week of ferrous fumarate supplementation, the Crohn Disease Activity Index tended to increase (P = 0.071). Patients experienced aggravation of diarrhoea, abdominal pain and nausea. Plasma-reduced cysteine was lower (P = 0.038) in patients than it was in controls. One week of ferrous iron supplementation further decreased reduced cysteine (P < 0.001) and significantly decreased plasma-reduced glutathione (P = 0.004) in the patients. Serum iron increased significantly in patients after an oral iron load test (from 5.8 +/- 3.2 micromol/L to 30.9 +/- 13.1 micromol/L). CONCLUSIONS: Treatment of iron deficiency with ferrous fumarate deteriorated plasma antioxidant status and increased specific clinical symptoms in patients with active Crohn disease. Plasma reduced cysteine may be a sensitive indicator for oxidative stress in the intestine.

Anemia in pregnancy in rural Tanzania: associations with micronutrients status and infections.

OBJECTIVE: We studied the association between anemia in pregnancy and characteristics related to nutrition and infections. DESIGN: Cross-sectional study. SETTING: Four antenatal clinics in rural northern Tanzania. SUBJECTS/METHODS: A total of 2547 women were screened for hemoglobin (Hb) and malaria plasmodia in capillary blood and for infections in urine. According to their Hb, they were assigned to one of five groups and selected accordingly, Hb<70 g/l (n=10), Hb=70-89 g/l (n=61), Hb=90-109 g/l (n=86), Hb=110-149 g/l (n=105) and Hb> or =150 g/l (n=50). The 312 selected subjects had venous blood drawn, were interviewed, and their arm circumference was measured. The sera were analyzed for ferritin, iron, total iron binding capacity (TIBC), cobalamin, folate, vitamin A, C-reactive protein (CRP), and lactate dehydrogenase (LD). Transferrin saturation (TFsat) was calculated. Urine was examined by dipsticks for nitrite. MAIN OUTCOME MEASURES: Unadjusted and adjusted odds ratio (OR and AOR) of anemia with Hb<90 g/l. RESULTS: Anemia (Hb<90 g/l) was associated with iron deficiency (low s-ferritin; AOR 3.4). The association with vitamin deficiencies were significant in unadjusted analysis (low s-folate; OR 3.1, low s-vitamin A; OR 2.6). Anemia was also associated with markers of infections (elevated s-CRP; AOR 3.5, urine nitrite positive; AOR 2.4) and hemolysis (elevated s-LD; AOR 10.1). A malaria positive blood slide was associated with anemia in unadjusted analysis (OR 2.7). An arm circumference less than 25 cm was associated with anemia (AOR 4.0). The associations with less severe anemia (Hb 90-109 g/l) were similar, but weaker. CONCLUSIONS: Anemia in pregnancy was associated with markers of infections and nutritional deficiencies. This should be taken into account in the management of anemia at antenatal clinics. SPONSORSHIP: The study was supported by the Norwegian Research Council (NFR) and the Centre for International Health, University of Bergen.

Iron supplementation in pregnancy--evidence and controversies.

Approximately 20% of women in industrialized countries have iron deficiency in pregnancy. This article focuses on the diagnostic problem of anemia and iron deficiency and discusses different strategies for iron supplementation in pregnancy. S-ferritin is commonly used to diagnose empty iron stores and is considered useful early in pregnancy as a diagnostic tool. Mean cellular volume (MCV), s-Fe and erythrocyte distribution width is too unspecific. Serum transferrin receptor (sTfR) is a relatively novel promising indicator of iron deficiency. Iron demands of the pregnant women are discussed as well as the dietary content of iron. Both beneficial and adverse effects of iron supplementation are outlined. It is not documented that supplementation has any substantial effect on birth weight or various complications in pregnancy. However, supplementation corrects the iron store and biochemical parameters of iron deficiency including hemoglobin concentration (Hb) and maintains the maternal iron stores in the puerperium. Recent literature also suggests that iron supply to the pregnant women may have beneficial effects on the iron content of neonates the first year of life.

Exposure to hyperoxia in diving and hyperbaric medicine--effects on blood cell counts and serum ferritin.

A reduction in hemoglobin concentration has been consistently reported after deep saturation dives, whereas reductions in thrombocyte counts and changes in biochemical parameters specific for liver function have been reported after some dives. In this study the contribution of exposure to hyperoxia to these changes were studied. Hemoglobin concentration, blood cell counts, serum ferritin, and biochemical parameters specific for liver damage were measured before and after a shallow 28-day saturation dive to a pressure of 250 kPa with the same hyperoxic exposure (40-50 kPa) as in a deep saturation dive in eight male divers. The same parameters were measured before, during, and after a standard 21-day hyperbaric oxygen (HBO2) treatment series in a selected group of 16 patients (8 male). There were significant reductions in hemoglobin concentrations of 3.8 +/- 4.7% (P = 0.023) and 10.2 +/- 5.3% (P = 0.003) after the HBO2 treatment series and dive, respectively, accompanied with reductions in red cell counts, reticulocyte counts, and hematocrit. There was an increase in ferritin concentrations of 29 +/- 21% (P = 0.002) and 107 +/- 43% (P < 0.001). In contrast to some deep dives, there were no changes in thrombocyte counts or biochemical parameters specific for liver damage. Exposure to hyperoxia contributes significantly to reduced hemoglobin and increased ferritin concentrations after saturation dives. The changes may reflect a shift of iron from synthesis of hemoglobin in the bone marrow to storage in macrophages caused by a downregulation of hemoglobin synthesis, or an increased oxidative stress. The changes are too small to be of clinical significance with respect to diving and HBO2 treatment.

Trace element reference values in serum determined by inductively coupled plasma atomic emission spectrometry.

Serum reference values for Ba, B, Cd, Cu, Fe, Mn, Li, Se, Sr, and Zn in 141 healthy Norwegians were determined. The trace element concentrations were determined by the inductively coupled plasma atomic emission spectrometry technique that we have recently validated. The reference intervals were established according to the recommendations of the International Federation of Clinical Chemistry and Laboratory Medicine. Also coverage intervals with coverage uncertainties were calculated according to the International Union of Pure and Applied Chemistry. The population studied consisted of 69 men and 72 women of the ages 21-87 years. The effects of gender, age, smoking, and oral contraceptives on serum levels of trace elements were investigated. Median concentrations of the different trace elements in (micromol/l) were as follows: Ba (0.44), B (1.50), Cd (0.004), Cu (17.1), Fe (21.4), Li (0.06), Mn (0.003), Se (1.26), Sr (0.17), and Zn (13.3). An increase in serum Ba and Sr was detected with age. These metals and Se were also significantly higher in women over 50 years of age in comparison to younger women. Women had higher serum Cu than men and those on oral contraceptives had higher serum Cu and Fe. Serum B tended to increase with age, while it was significantly reduced with smoking.

Anaemia in pregnancy: possible causes and risk factors in Nepali women.

OBJECTIVE: The aim of this study was to investigate the importance of nutritional deficiencies and infections in the development of anaemia in pregnant Nepali women. DESIGN: Case-control study. SETTING: Patan Hospital, Kathmandu, Nepal. SUBJECTS: A sub-sample (n=479) of all pregnant women (n=2856) coming for their first antenatal visit in a 12 month period, 1994-1995. Women who had already received any micronutrient supplementation (n=82), and those whose serum samples showed macroscopic haemolysis (n=7) were excluded. The remaining women (n=390) were included in the statistical analysis. They were divided into three groups; a non-anaemic control group, haematocrit (Hct)>33% (n=82), and two case-groups: moderately anaemic, Hct 25-33% (n=254), and severely anaemic, Hct<25% (n=54). RESULTS: We found high prevalences of nutritional deficiencies and intestinal infections, both among cases and controls. The prevalence of low s-ferritin was high, especially among the severely anaemic women (55.6%). In a multiple logistic regression model, the presence of low s-vitamin A, elevated s-C-reactive protein or hookworm infection was associated with a significantly increased risk of severe anaemia. The adjusted odds ratios (95% CI) were 8.38 (1.99, 35.30), 4.91 (1.22, 19.67) and 5.43 (1.20, 24.61), respectively. CONCLUSIONS: In addition to the present routine iron and folate supplementation to pregnant Nepali women, vitamin A supplementation needs to be considered. Prevention and treatment of infections should, together with dietary advice, be emphasized more strongly in the antenatal care. SPONSORSHIP: The Norwegian Research Council and the Norwegian Universities Committee for Development, Research and Education. European Journal of Clinical Nutrition (2000) 54, 3-8

Validation of inductively coupled plasma atomic emission spectrometry technique (ICP-AES) for multi-element analysis of trace elements in human serum.

The use of inductively coupled plasma atomic emission spectrometry (ICP-AES) for the simultaneous determination of Al, B, Ba, Be, Cd, Co, Cr, Cu, Fe, Li, Mn, Ni, Pb, Se, Sr and Zn in human serum in a clinical laboratory was validated. Samples were digested and then analysed using yttrium as an internal standard and a serum-matched calibration standard. The criteria used to assess the analytical performance of the ICP-AES were detection and quantification limits, linearity, sensitivity, recovery, interference from alkali and acid, trueness and precision. Detection limits were 0.002-0.003 micromol/L for Mn, Sr, Ba, and Cd; 0.014-0.07 micromol/L for Co, Zn, Fe, Be, Li, Pb, Cu, Ni, and Cr; and 0.2-0.9 micromol/L for B, Se, and Al. Trueness, as controlled by analysis of bovine serum certified reference material, was acceptable for Co, Cu, Se and Zn, while Fe was 5.1% and Mn 6.2% below the lowest limit of the certified material interval. We conclude that ICP-AES can be used for multi-element analysis of B, Ba, Cu, Fe, Li, Se, Sr and Zn in serum. Serum levels of Al, Be and Co were below the detection limits while serum levels of Cd, Cr, Ni and Pb were below the quantification limits of the ICP-AES. These trace metals cannot be analysed as routine by the ICP-AES. However, in cases of intoxication with elevated serum concentrations mean recovery of 100+/-10% was obtained at an addition of 2.22 micromol/L for Al, 0.11 micromol/L for Be, 0.03 micromol/L for Co, 0.39 micromol/L for Cr, 0.14 micromol/L for Ni, and 0.12 micromol/L for Pb.

[Bloodletting as medical therapy for 2500 years]. / Arelating som medisinsk behandling i 2500 år.

Bloodletting has been part of the history of medicine for more than 2500 years. Up to the end of the Middle Ages, the rationale for bloodletting originated from the ancient greek humoral theory. The great scientific progress from the 16th century and onward, apparently did not weaken its position. Prominent physicians such as Andreas Vesalius (1514-64), William Harvey (1578-1657) and Thomas Sydenham (1624-89) defended bloodletting. In the beginning of the 19th century the use of leeches became the major technique of bloodletting in Europe. In Norway bloodletting was mentioned in royal decrees from the 13th century, and the method became popular in folk medicine. At the end of the 19th century bloodletting came at last to be regarded as ineffective for most of its traditional purposes, and its use declined rapidly. Today, however, bloodletting is being restored in modern medicine as the most effective method of treating the increasing frequent disorders caused by iron overload.