In August 2002, the Subcommittee on Cessation of the Interagency Committee on Smoking and Health (ICSH) was charged with developing recommendations to substantially increase rates of tobacco cessation in the United States. The subcommittee's report, A National Action Plan for Tobacco Cessation, outlines 10 recommendations for reducing premature morbidity and mortality by helping millions of Americans stop using tobacco. The plan includes both evidence-based, population-wide strategies designed to promote cessation (e.g., a national quitline network) and a Smokers' Health Fund to finance the programs (through a 2 US dollar per pack excise tax increase). The subcommittee report was presented to the ICSH (February 11, 2003), which unanimously endorsed sending it to Secretary Thompson for his consideration. In this article, we summarize the national action plan.

Hemoglobinometry according to the International Committee of Standardization in Hematology (ICSH) suffers from imprecision related to high sample dilution and from potential errors owing to sample turbidity. We have evaluated a new instrument, "HemoCue," that measures hemoglobin at two wavelengths as azide methemoglobin, without dilution. The HemoCue method is superior to the ICSH method: by correction for turbidity, it avoids false hemoglobin readings that may arise from hyperlipemia or some large M-component of the immunoglobulin M class. We find the equipment suitable for use in outpatient units.

Using either a bone-ligament-bone or a muscle-tendon-bone preparation, numerous investigators have demonstrated that the usual site of separation is in the transitional zone between the ligament (or tendon) and bone; hence, the term junction strength or load at separation is used to describe functional changes in these preparations. Junction strength is decreased with inactivity (immobilization) and increased with chronic activity (training) provided that the exercise program is of an endurance nature. Training also increases junction strength in thyroidectomized and hypophysectomized rats. Besides in junction strength, training results in heavier ligaments and higher ligament weight/length ratios. However, water content, collagen concentrations/dry weight or collagen concentration per weight/length unit are not significantly influenced by repeated bouts of exercise. Although immobilization is associated with lower elastic stiffness values (kg/mm), training appears to have little influence on this measure in normal animals. Rats and dogs with surgically repaired ligaments are weaker and the strength results are markedly lower if the leg is immobilized. Exercise training improves the repair strength of ligaments but does not result in normal values twelve weeks after the surgery. Exogenous administration of ICSH or testosterone results in higher repair strength whereas TSH, thyroxine, ACTH and growth hormone decreases this measure. It was concluded that the mechanical stress produced by chronic exercise is an important determination of the strength of repaired ligaments and of the junctions between ligaments (or tendons) and bones.

BACKGROUND

According to WHO 2008 guidelines, the required percentage of cells manifesting dysplasia in the bone marrow to qualify as significant is 10% or over in one or more hematopoietic cell lineages, but this threshold is controversial. No 'normal' values have been established. Therefore, we investigated dyshematopoiesis in bone marrow aspirate squash preparations of 120 healthy bone marrow donors.

METHODS

Bone marrow squash slides of 120 healthy unrelated bone marrow donors were examined independently by 4 experienced morphologists. Samples were taken from the first aspiration during the harvest. Bone marrow preparation and assessment were performed according to WHO recommendations and ICSH guidelines.

RESULTS

More than 10% dysmyelopoiesis could be detected in 46% of bone marrow aspirate squash preparations with 26% in 2 or more cell lineages and 7% in 3 cell lineages in healthy bone marrow donors. Donors under the age of 30 years exhibited more dysgranulopoietic changes and dysmegakaryopoietic changes (P<0.001) compared to the older donors. Female donors showed more dysgranulopoietic changes than male donors (P = 0.025). The concordance rate between the 4 investigators was modest in dysgranulopoiesis but poor in dyserythropoiesis and dysmegakaryopoiesis.

CONCLUSIONS

The poor reliability of the 10% cut off was partly related to the proximity of the current criteria to the observed cut-off mean values of the normal population. These findings question the current WHO threshold of the 10% or over necessary for the percentage of cells manifesting dysplasia to be considered significant, and suggest that either a higher threshold would be more appropriate or different thresholds should be set for each lineage.

The Expert Panel on Blood Rheology of the International Council for Standardization in Haematology (ICSH) has prepared new recommendations for measurement of the erythrocyte sedimentation rate (ESR) under the following categories: 1. ICSH reference method-ICSH now recognises, as its reference method for the ESR, the sedimentation of EDTA-anticoagulated but undiluted blood in traditional Westergren pipettes that meet ICSH specifications. 2. ICSH standardised method-ICSH recommends specifications for a new standardised method for the ESR based on the sedimentation of EDTA-anticoagulated, but undiluted blood in pipettes with a 200 mm scale and which are designed to avoid spillage of blood or aerosol generation. This standardised method may be used for verification or quality control of other ESR methods and, in future, may replace the reference method. 3. ICSH selected methods-ICSH recommends specifications for working methods, using diluted or undiluted blood, which may be considered as ICSH selected methods for routine use. A protocol is outlined for evaluation of such working methods against the ICSH reference method or the new ICSH standardised method.

In hypophysectomized immature female rats (HIFR), the ovarian weight response to subcutaneously implanted diethylstilbestrol capsules (DESC) is inhibited by small doses of human chorionic gonadotropin (hCG). This effect, reproduced by equivalent doses of interstitial cell stimulating hormone (ICSH) but not by follicle-stimulating hormone (FSH), is inhibited by treatment with antiandrogens. These data implicate gonadotropic stimulation of interstitial cell androgen production in the control of follicular maturation in rats.

Schistocytes are fragments of red blood cells (RBCs) produced by extrinsic mechanical damage within the circulation. The detection of schistocytes is an important morphological clue to the diagnosis of thrombotic microangiopathic anemia (TMA). Reporting criteria between different laboratories, however, are not uniform, owing to variability of shape and nature of fragments, as well as subjectivity and heterogeneity in their morphological assessment. Lack of standardization may lead to inconsistency or misdiagnosis, thereby affecting treatment and clinical outcome. The Schistocyte Working Group of the International Council for Standardization in Haematology (ICSH) has prepared specific recommendations to standardize schistocyte identification, enumeration, and reporting. They deal with the type of smear, method of counting, morphological description based on positive criteria (helmet cells, small, irregular triangular, or crescent-shaped cells, pointed projections, and lack of central pallor). A schistocyte count has a definite clinical value for the diagnosis of TMA in the absence of additional severe red cell shape abnormalities, with a confident threshold value of 1%. Automated counting of RBC fragments is also recommended by the ICSH Working Group as a useful complement to the microscope, according to the high predictive value of negative results, but worthy of further research and with limits in quantitation.

The effect of lyophilized aqueous extract of Cynomorium coccineum and Withania somnifera on testicular development and on serum levels of testosterone, ICSH and FSH was studied in immature male Wistar rats. There was a notable increase in testicular weight of animals treated with both extracts. Histological examination revealed an apparent increase in the diameter of seminiferous tubules and the number of seminiferous tubular cell layers in the testes of treated rats as compared with control ones. Extracts of both plants elicited notable spermatogenesis in immature rats but C. coccineum was more effective than W. somnifera in that respect. Serum testosterone and FSH levels were lower in animals treated with plants extracts than controls, whereas ICSH levels was higher in treated animals, specially in those treated with C. coccineum. It was concluded that extracts of both plants have a direct spermatogenic influence on the seminiferous tubules of immature rats presumably by exerting a testosterone-like effect.

In recognition of the need for a standardization of the measurement of the erythrocyte sedimentation rate (ESR), the International Council for Standardization in Haematology makes the following recommendations: (i) The reference method for measurement of the ESR should be based on the Westergren method, which is a specific test for the ESR, with modifications, (ii) The reference method for measurement of the ESR should use either whole blood anticoagulated with EDTA and later diluted with sodium citrate or saline (4 : 1) or whole blood anticoagulated with sodium citrate (4 : 1) in Westergren pipettes, (iii) The ESR pipettes can be of glass or plastic (with specific characteristics). It must be colourless; a minimum sedimentation scale of 200 mm, a minimum bore of 2.55 mm, which should be constant within 5%. A protocol for the evaluation of alternative methodologies against the reference method is outlined: The new technologies must be tested over a range of ESR values of 2-120 mm. In this comparison, 95% of the differences should be 5 mm or less, with larger differences associated with higher ESR values. A minimum of 40 samples should be tested in 3 different groups of values: 1-20, 21-60 and more than 60 mm. The statistical methods recommended for ESR evaluations are the coefficient of correlation, the Passing-Bablock regression and the Bland-Altman statistical method. This reference method replaces all earlier standardized and reference methods.

The bone marrow examination is an essential investigation for the diagnosis and management of many disorders of the blood and bone marrow. The aspirate and trephine biopsy specimens are complementary and when both are obtained, they provide a comprehensive evaluation of the bone marrow. The final interpretation requires the integration of peripheral blood, bone marrow aspirate and trephine biopsy findings, together with the results of supplementary tests such as immunophenotyping, cytogenetic analysis and molecular genetic studies as appropriate, in the context of clinical and other diagnostic findings. Methods for the preparation, processing and reporting of bone marrow aspirates and trephine biopsy specimens can vary considerably. These differences may result in inconsistencies in disease diagnosis or classification that may affect treatment and clinical outcomes. In recognition of the need for standardization in this area, an international Working Party for the Standardization of Bone Marrow Specimens and Reports was formed by the International Council for Standardization in Hematology (ICSH) to prepare a set of guidelines based on preferred best practices. The guidelines were discussed at the ICSH General Assemblies and reviewed by an international panel of experts to achieve further consensus.

OBJECTIVE

Reliable blood donor screening requires more accurate measure of haemoglobin (Hb) than by either copper sulphate or the haemoglobin colour scale. The HemoCue haemoglobinometer has established a method for this, but it is considerably more expensive; a modified version (HemoCue 301) has now been developed with a cheaper reagent-free cuvette for use in budget-restricted situations. This report describes evaluation of the performance, the assessment of reproducibility and accuracy of this modified analyser against the reference technique for Hb measurement.

METHODS

Over 300 routine blood samples from specimens received routinely in a hospital laboratory were tested in accordance with the International Committee for Standardization in Haematology (ICSH) protocol. Accuracy and linearity were confirmed by the reference method with the WHO international haemoglobincyanide reference standard. Tests were also performed on selected samples for checking interference by biochemical abnormalities and leucocytosis. The effects of various sample storage conditions prior to testing were also tested.

RESULTS

Ninety per cent of results were within 4% of true values, 96% within 6% and in only three cases was the deviation>> 10%, due to interference by bilirubinaemia and/or C-reactive protein. At an Hb value of 120 g/l for donor selection, there were no cases where the method would have been misleading.

CONCLUSIONS

HemoCue 301 provides a simple and reliable anaemia screen method, conforming to the requirements of CLIA'88 regulations; it is reliable for discriminating Hb values for donor acceptance. The main advantage is that the cuvettes are significantly cheaper than the previous models, and will not deteriorate in adverse climatic conditions.

BACKGROUND

Hereditary spherocytosis (HS), hereditary elliptocytosis (HE), and hereditary stomatocytosis (HSt) are inherited red cell disorders caused by defects in various membrane proteins. The heterogeneous clinical presentation, biochemical and genetic abnormalities in HS and HE have been well documented. The need to raise the awareness of HSt, albeit its much lower prevalence than HS, is due to the undesirable outcome of splenectomy in these patients.

METHODS

The scope of this guideline is to identify the characteristic clinical features, the red cell parameters (including red cell morphology) for these red cell disorders associated, respectively, with defective cytoskeleton (HS and HE) and abnormal cation permeability in the lipid bilayer (HSt) of the red cell. The current screening tests for HS are described, and their limitations are highlighted.

RESULTS

An appropriate diagnosis can often be made when the screening test result(s) is reviewed together with the patient's clinical/family history, blood count results, reticulocyte count, red cell morphology, and chemistry results. SDS-polyacrylamide gel electrophoresis of erythrocyte membrane proteins, monovalent cation flux measurement, and molecular analysis of membrane protein genes are specialist tests for further investigation.

CONCLUSIONS

Specialist tests provide additional evidence in supporting the diagnosis and that will facilitate the management of the patient. In the case of a patient's clinical phenotype being more severe than the affected members within the immediate family, molecular testing of all family members is useful for confirming the diagnosis and allows an insight into the molecular basis of the abnormality such as a recessive mode of inheritance or a de novo mutation.

Blood rheology is the science of the flow and deformation of blood. Clinically, blood rheology is important because circulatory resistance has two major components, vascular and rheological. In large vessels, blood rheology should be considered in terms of bulk flow, the viscosity of blood depending mainly on red cell concentration and plasma viscosity and, to a lesser extent, on red cell deformability and aggregation. In the microcirculation, where cells must deform to pass through narrow capillaries, cellular rheology (i.e. the deformability of individual cells) is a major determinant of resistance to flow. This ability to deform is also a determinant of the cell's survival time in the circulation. The deformability of the red cell is essentially linked to its structure (i.e. its cellular geometry, membrane properties and cytoplasmic viscosity); thus structural abnormalities, as found in some haematological disorders, can be expected to affect blood flow in the microcirculation and/or red cell lifespan. Blood rheology is a relatively new discipline as applied to the practice of haematology. In 1985 the International Committee for Standardization in Haematology (ICSH) established an Expert Panel on Blood Rheology which has subsequently issued guidelines on the measurement of blood viscosity and erythrocyte deformability and on tests such as erythrocyte sedimentation rate and plasma viscosity that are used to monitor the acute phase response in inflammatory disease. Rheological methods now have sufficiently good sensitivity and specificity for their application to a wide variety of clinical disorders. This review illustrates their potential application to haematological disorders that cause abnormal deformability of red cells.

Although DNA analysis is needed for characterization of the mutations that cause β-thalassaemia, measurement of the Hb A(2) is essential for the routine identification of people who are carriers of β-thalassaemia. The methods of quantitating Hb A(2) are described together with pitfalls in undertaking these laboratory tests with particular emphasis on automated high-performance liquid chromatography and capillary electrophoresis.

This guidance document was prepared on behalf of the International Council for Standardization in Haematology (ICSH) for providing haemostasis-related guidance documents for clinical laboratories. This inaugural coagulation ICSH document was developed by an ad hoc committee, comprised of international clinical and laboratory direct acting oral anticoagulant (DOAC) experts. The committee developed consensus recommendations for laboratory measurement of DOACs (dabigatran, rivaroxaban, apixaban and edoxaban), which would be germane for laboratories assessing DOAC anticoagulation. This guidance document addresses all phases of laboratory DOAC measurements, including pre-analytical (e.g. preferred time sample collection, preferred sample type, sample stability), analytical (gold standard method, screening and quantifying methods) and post analytical (e.g. reporting units, quality assurance). The committee addressed the use and limitations of screening tests such as prothrombin time, activated partial thromboplastin time as well as viscoelastic measurements of clotting blood and point of care methods. Additionally, the committee provided recommendations for the proper validation or verification of performance of laboratory assays prior to implementation for clinical use, and external quality assurance to provide continuous assessment of testing and reporting method.

The Westergren erythrocyte sedimentation rate (ESR) has been used for many years without any formal procedure for method validation or for quality assurance. In 1988, the International Committee on Standardization in Hematology (ICSH) (Leuven, Belgium) described an ESR validation procedure as well as a method for producing ESR reference material (ICSH reference method) in the laboratory where it is to be used. The ICSH proposal was tested in our laboratory during a consecutive period of 36 months. In this article, a new mathematical relationship between the ICSH recommendation and the Westergren method is developed, which can be easily used for method validation and/or quality assurance. A table has been made that establishes 95% action limits for Westergren ESRs based on ICSH reference ESRs from 5 through 105 mm/h. The table, derived from 36 months of data, has been tested against two new data sets and used to validate two commercial ESR methods. Analysis of outliers was performed with special attention to the mixing of whole blood samples with sodium citrate necessary for the Westergren ESR. This mixing process is best performed in a large-bore test tube as opposed to the use of the Westergren ESR tube as an aliquoting and mixing device.

Multi-color flow cytometry is a unique technology, which enables the analysis of heterogeneous cellular systems and provides multiparametric information on a cell-by-cell basis. A variety of factors contribute to the complexity of validating cell-based flow cytometric methods, including the lack of fully characterized cellular reference materials and the difficulty in obtaining, or creating, samples with varying levels of a given cell type or varying levels of expression of a given antigen. This document summarizes validation requirements and describes validation strategies for quasi-quantitative and qualitative cell-based flow cytometric assays.