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At least 217 records · Page 12Linked to original sources
PubMed1986

Basic principles and problems of haemocytometry.

After some brief remarks on counting chambers, references to the ICSH-recommended haemoglobin-determination are given. The microhaematocrit of normal blood is advocated as a potential routine calibration method. Comments are given on discrepancies between centrifugal and flow haemocytometry haematocrits of abnormal and artificial bloods. Flow haemocytometry instruments are classified into analogue and digital instruments or into electrical and optical instruments. Their hydrodynamic properties are discussed. The principles and problems of electrical and optical cell counting and sizing are dealt with. The importance of the refractive index and of flow-induced cell shape changes for the MCV determinations is stressed. It is argued that MCV and haematocrit values are exaggerated at both low and high values and consequently MCHC is erroneously constant. Various prevailing red cell distribution width (RDW) and platelet distribution width (PDW) definitions bring about considerable confusion. The major features of the counting and sizing of white blood cells and platelets are described.

Blood Cell Count↗
PubMed1975

Effects of human chorionic gonadotropin, human interstitial cell stimulating hormone and human follicle-stimulating hormone on ovarian weights in estrogen-primed hypophysectomized immature female rats.

In connection with systematic studies of steroid and peptide hormone interactions during follicular growth, we have measured ovarian weight responses to graded doses of highly purified human chorionic gonadotropin (hCG), human interstitial cell stimulating hormone (hICSH)in hypophysectomized immature female rats (HIFR) treated with diethylstilbestrol in silastic capules (desc) implanted subcutaneously. Our results are consistent with earlier reports of enhancement of ovarian weight responses to hCG and FSH. Contrary to results of similar experiments reported by others, we have found that estrogen treatment of HIFR enhanced ovarian weight response to ICSH. In addition, we report for the first time that small doses of hCG and hICSH inhibit ovarian weight responses to estrogen in HIFR. Our observations on effects of small doses of hCG and hICSH and the long-known fact that ovarian interstitial cells are stimulated in HIFR given similar doses of these hormones lead us to hypothesize that ovarian interstitial cell stimulation is involved in the control of follicular maturation.

Animals↗
PubMed1975

Evidence of a role of adrogens in follicular maturation.

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.

Androgens↗
PubMed2001

Platelet counting by the RBC/platelet ratio method. A reference method.

The International Council for Standardization in Haematology (ICSH) and the International Society of Laboratory Hematology (ISLH) recommend the counting of specifically labeled platelets relative to the RBCs with a fluorescence flow cytometer, together with an accurate RBC count determined with a semiautomated, single-channel aperture-impedance counter as a reference method for the enumeration of platelets. Fresh EDTA-anticoagulated venous blood specimens are measured within 4 hours of the draw. The specimen is prediluted (1:20) and the platelets labeled with two monoclonal antibodies specific to a cluster of differentiation common to all platelets. A final 1:1,000 dilution is made and at least 50,000 events with a minimum of 1,000 platelet events are counted with a flow cytometer to determine the RBC/platelet ratio. The platelet count is then calculated from this ratio and the RBC concentration of the original blood specimen.

Anticoagulants↗
PubMed1989

Evaluation of the automated haematology analyser Sysmex M-2000.

The automated Sysmex M-2000 was evaluated according to the ICSH (International Committee for Standardization in Haematology) protocol. After dilution of packed cells with cell-free plasma, blood cell counts were linear. The overall precision of the measured parameters was good; the CV's ranged between 0.64% and 2.06%. The carry-over was negligible; platelets showed the biggest carry-over with 0.25% in the Whole Blood Mode, while red blood cells (RBC) showed a carry-over of 0.55% in the Prediluted Mode. 300 clinical samples were measured on the Sysmex M-2000 and the Sysmex CC-700 with PL-100, and the results were compared. The coefficients of correlation for white blood cells (WBC), red blood cells (RBC), haemoglobin and haematocrit were greater than 0.99; platelets showed an r of 0.982. Comparison of the results from the Sysmex M-2000 trimodal leukocyte distribution with a manual 100 cell differentiation showed a close correlation for lymphocytes (r = 0.948), and neutrophils (r = 0.931). The middle cell fraction corresponding to monocytes, eosinophils and basophils showed a correlation with r = 0.703. Pathological samples showed no interference with the blood count. Leukocyte counts less than 1000 x 10(9)/l did not effect the measurement of haemoglobin. During the period of evaluation, no instrument malfunctions occurred. Because of its precision and reliability, the Sysmex M-2000 is well suited for routine work and stat analysis in medium-sized laboratories.

Blood Cell Count↗
PubMed2001

Length of sedimentation reaction in undiluted blood (erythrocyte sedimentation rate): variations with sex and age and reference limits.

Although the length of sedimentation reaction in blood (LSRB) (commonly, but improperly called erythrocyte sedimentation rate, ESR) has long been used in clinical laboratories because it is simple and low-cost, its sensitivity and specificity are unsatisfactory. Usually, the values are reported using the Westergren method with sodium citrate-anticoagulated specimens. We used a new procedure, the TEST1, which measures the length of sedimentation reaction in undiluted K3EDTA anticoagulated blood samples following ICSH (International Committee for Standardization in Haematology) recommendations. Samples obtained from 840 reference individuals (430 females and 410 males, mean age 44 and 46.5 years respectively, range 1 to 90 years) were utilised to estimate the reference limits. The subjects, classified by sex, were subdivided into four statistically different age groups to determine the reference limits (2.5th and 97.5th percentiles). Sex difference was statistically significant in two age groups, from 14 to 50 (p < 0.0001) and from 50 to 70 years (p < 0.01). We did not observe significant sex difference within the age bracket from 1 to 14 years and from 70 to 90 years. In both sexes LSRB values increased with age, in significant correlation with fibrinogen concentration (p < 0.0001), and became significantly higher in subjects older than 70 compared to all the younger subjects (p<0.01 in females and p < 0.02 in males). Thus, we defined adequate reference ranges in elderly.

Adolescent↗
PubMed1975

The use of antiserum to rat prolactin to evaluate its role as luteotrophin in the hamster.

Rabbit antisera to rat prolactin was found to cross-react with the hamster pituitary extract by double diffusion and quantitative precipitin techniques. Short term neutralization of prolactin by antiserum to rat prolactin either in early or late stages of gestation did not have any deleterious effects. Prolonged administration of rat prolactin antiserum resulted in an increase in the incidence of resorptions, but pregnancy was still maintained followed by normal parturition. This is in marked contrast to the abortive effects of an antiserum of ICSH. It was concluded that prolactin does not play a major role in maintaining pregnancy of the hamster.

Animals↗
PubMed1977

Biologically active luteinizing hormone (LH) in plasma: II. Comparison with immunologically active LH levels throughout the human menstrual cycle.

The levels of biologically active luteinizing hormone were determined by an in vitro bioassay method in plasma samples collected daily over a complete menstrual cycle from 12 menstruating women. These cycles were normal according to a number of criteria, including daily plasma levels of oestradiol, 17-hydroxyprogesterone and progesterone. Immunoreactive LH was estimated in the same 12 cycles by a radio-immunoassay (RIA) procedure (HCG-RIA) using an HCG antiserum and iodinated HCG. The 2nd IRP of HMG was selected as standard although significant deviations from parallelism were found with 7 out of the 12 plasma pools studied. The use of the 1st IRP of human pituitary gonadotrophins (FSH and LH (ICSH)) for bioassay (herafter HPG-1st IRP) as standard in this system resulted invariably in invalid assays, due to lack of parallelism. Immunoreactive LH was also measured in 8 of the 12 cycles by a RIA procedure (HLA-RIA) using a human LH antiserum and iodinated human LH of pituitary origin. Results are expressed in terms of the HPG-1st IRP. The plasma levels of biologically and immunologically active LH were qualitatively similar throughout the menstrual cycle. However, the LH levels measured by the bioassay invariably exceeded those estimated by the RIA procedures. The biological to immunological (B/I) ratio over the entire menstrual cycle (312 comparisons) was 5.5 with 95% confidence limits at 5.2 and 5.8 when the HCG-RIA system was employed. Using the HLH-RIA system (208 comparisons), the corresponding ratio was 6.4 (6.0:6.9). When regression lines were calculated using the bioassay results as the independent variable and the RIA results as the dependent variable, the 95% confidence limits of the regression lines did not include the origin. Furthermore,, in keeping with the high B/I ratios, the slopes of the two regression lines and their conficence limits differed markedly from unity. It is concluded that although qualitatively similar profiles were observed between the biological and immunological activities throughtout the menstrual cycle, two aspects require further attention. Firstly, the elevated B/I ratios together with the behaviour of the dose-effect lines obtained with different standards in the various RIA systems suggest that presently available reference standard preparations of pituitary and/or urinary origin are not suitable for the assay of LH in human plasms. Secondly, from the regression analyses of the biological and immunological activities it is infered that the RIA methods detect immunological activity which is not associated with biological activity. If so, the validity of these RIA procedures for specifically measuring low levels of biologically active LH in plasma may be in question.

Adult↗
PubMed1979

Effect of storage on the bioactivity of two international reference preparations of human pituitary luteinizing hormone in vitro.

There was no loss of biological activity in vitro when the 1st International Reference Preparation (IRP) for human pituitary gonadotrophins [FSH and LH/interstitial cell stimulating hormone (ICSH) for bioassay] code no. 69/104 and the 1st IRP for human pituitary LH/ICSH [for immunoassay] code no. 68/40 were stored for 1 year at -70 degrees C in a buffered 0.8% saline solution containing 1% bovine plasma albumin (BPA). However, storage of the 69/104 preparation at -20 degrees C in either 0.1 or 1% BPA, or at -70 degrees C in the presence of 0.1% BPA showed a small but significant decrease (approximately 10%) in activity over the same period. It is, therefore, advantageous to store these reference preparations at -70 degrees C in the presence of 1% BPA.

Drug Storage↗
PubMed1980

Haemoglobin determination without cyanide on Hemalog-8.

The most common method for determination of haemoglobin is the cyanmet-haemoglobin method which is recommended by the International Committee for Standardization in Haematology (ICSH). Determination of haemoglobin is one of the most used methods in hospital laboratories. One disadvantage with the method is the use of cyanide in the reagent, which demands cautious preparation and use. The intention of our investigation was to replace the cyanide reagent for Hemalog-8 with a reagent not containing cyanide. It is shown that the change of reagent did not influence the results of the analysis.

Cyanides↗
PubMed1980

Pyruvate kinase variants characterized by the methods recommended by the International Committee for Standardization in Haematology.

Seven new pyruvate kinase (PK) variants were characterized by the recently established recommended methods by the International Committee for Standardization in Haematology (ICSH). The seven cases were all true homozygotes as evidenced by consanguineous marriages of the parents. All of them were Japanese. These variants were designated PK Tokyo, Nagasaki, Sapporo, Maebashi, Itabashi, Fukushima, and Aizu. Low substrate affinity for phosphoenolpyruvate and thermal instability seem to play a major role in causing clinical manifestation of chronic hemolysis. Product inhibition of PK by ATP may also play an additional role in some cases.

Adolescent↗
PubMed1981

Histological changes in the testis of the domestic fowl after partial adenohypophysectomy.

The effects of partial adenohypophysectomy on comb size, testicular weight, and histology 20 or 30 days after the operation are described and discussed in relating these observed changes with the volume of remaining adenohypophyseal tissue. From the present observation, it is concluded that 1) the response of the domestic fowl to partial deficiency of adenohypophyseal tissue may be more sensitive than that of some mammals, 2) the decrease in testicular weight and tubular diameter precedes the histological change in testis and comb regression, and 3) to maintain fully active testicular function, more than 75% of the adenohypophysis is needed. About 40% of the adenohypophyseal tissue including a remnant of cephalic lobe can stimulate spermatogenesis, but the spermatozoa produced were partially abnormal. The critical amount of adenohypophyseal tissue for normal spermiogenesis may exist between 55 and 75%. All histological change is initially observed in the center of testis and then extended toward the periphery. It is suggested that the cephalic lobe contains at least a trace of ICSH in addition to FSH.

Animals↗
PubMed1975

[Determination of blood iron and the total fixation capacity of transferrin by a simple automatic technic without dialysis].

An automated method without dialysis for determining serum iron is described. It is an adaptation of the Lauber procedure to the Technicon Auto Analyser I: serum iron is split from its protein combination by exposure to an acetate pH 5.8 buffer, in the presence of a detergent (Teepol 710) which prevents any precipitation of proteins. Iron is maintained in ferrous state by addition of sodium dithionite and is quantitated with bathophenanthrolin colorimetry. Serum samples are analysed at a rate of 60/H. It is necessary to run a blank for each serum sample. Between 20 and 500 mug iron/100 ml the color intensity agrees with Beer's law. The reproducibility is good, and the variation coefficient less than 1.2 per cent. Our results are not significatively different of those obtained with the ICSH standard method. Bilirubin does not interfere. Iron added is satisfactorily recovered. Our method can also be used for determining the transferrin iron binding capacity.

Autoanalysis↗
PubMed1992

Evaluation of a new haematology analyser for whole blood count and full differential (NE-8000).

We evaluated the fully automated haematology analyser TOA Sysmex NE-8000 over a three-month period according to the ICSH protocol using as reference techniques a Coulter STKR counter and microscope examination for WBC differential and cell morphology. The NE-8000 employs aperture impedance to perform cell counts, with a sheath fluid to focus cells hydrodynamically prior to counting and sizing. WBC are differentiated into five populations. A combination of aperture impedance, radio frequency measurement and differential cell shrinkage is used, eosinophil and basophil percentages being established in two separate channels and subtracted from the total granulocyte count in order to give the value for neutrophils. Analysis of 1060 samples processed by the closed sampling automode demonstrated satisfactory counting performance. Among the WBC differentials obtained from 100 samples, neutrophil, eosinophil and lymphocyte results correlated well with those from microscopic examination of blood smears performed according to the NCCLS standard H20 T protocol. Differences observed in the percentages of basophils were of no biomedical significance. A comparative study for monocytes showed poor correlation for values below 5% and above 10%, best results being obtained in the intermediate range 5-10%. The NE-8000 also demonstrated good reliability for detection of abnormal cells.

Automation↗
PubMed2003

Performance evaluation of a new compact hematology analyzer, the Sysmex pocH-100i.

Medical practitioners are searching for ways to improve the quality and outcome of care while decreasing cost. One technological advance that may facilitate this is point-of-care testing. The pocH-100i hematology analyzer (Sysmex Corporation, Kobe, Japan) is a compact device designed specifically for point-of-care testing environments. The analyzer provides a full blood count and a 3-part differential leukocyte count. A distinct advantage is that neutrophils are reported separately, which is useful for monitoring oncology patients. The device was evaluated in accordance with International Council for Standardization in Haematology (ICSH) guidelines for precision, linearity, carryover, and effects of sample aging. The pocH-100i analyzer was compared with the Sysmex KX-21N device. The results for all parameters tested were almost identical. When samples including blast cells, immature granulocytes, and nucleated red blood cells were excluded, the pocH-100i automated differential compared well with a 400-cell manual differential. Results for neutrophils (r2 = 0.996), lymphocytes (r2 = 0.999), and the "mixed" population of cells (r2 = 0.611) indicated the pocH-100i analyzer would be highly suitable for low-volume laboratories and near-patient services.

Blood Cell Count↗
PubMed1992

Haemoglobin A2 measurement using high performance liquid chromatography.

Haemoglobin A2 levels were assessed using a modular high performance liquid chromatography (HPLC) system with a protocol designed for the measurement of haemoglobin A1C. There was good correlation (r = 0.96; P < 0.001) between this technique and the International Committee for Standardisation in Haematology (ICSH) recommended method of microchromatography. The range of haemoglobin A2 was found to be 2.3-3.2% in apparently normal individuals, and 4.0-6.7% in those with beta-thalassaemia trait. The HPLC system produced reliable results quickly for haemoglobin A2 with no alteration to the protocol used for measuring haemoglobin A1C.

Chromatography, High Pressure Liquid↗
PubMed2004

[Abnormalities of the erythrocyte membrane ultrastructure and the membrane proteins in a patient with HEMPAS, alpha thalassemia and complicated diabetes].

OBJECTIVE: Diagnosis of a case with congenital dyserythropoietic anemia (CDA). METHODS: Routine tests for hemolysis were carried out. The activities of erythrocyte enzymes were measured according to the methods recommended by international committee for standardization in hematology (ICSH). The quantity and quality of erythrocyte membrane proteins were analyzed with 4% - 15% sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis (SDS-PAGE). The membrane ultrastructure of erythrocyte from bone marrow was observed under transmission electron microscope (TEM). RESULTS: The main results were: (1) Bone marrow morphology revealed erythroid hyperplasia and 0.10 symmetrically binucleated late erythroblasts. The erythrocytes in peripheral blood showed anisocytosis and hypochromia. (2) The intracellular iron was 0.98 and the storage iron was strongly positive in bone marrow. The serum ferritin was 1607 micro g/L. The content of blood sugar was 27.5 mmol/L. (3) Ham test was negative in his own acidified serum but positive in the group-compatible sera. (4) A quick mobile H band was seen in hemoglobin electrophoresis. H inclusion test was positive. (5) SDS-PAGE demonstrated that the migration of band 3 protein of erythrocyte membrane in an electric field was faster (110%) than that of normal controls and the relative contents of band 1, band 3, band 4.1 were reduced to various extent. (6) "Double membrane" with gap and shedding was observes under TEM. CONCLUSIONS: The final diagnosis of the case was CDAII, also called HEMPAS (hereditary erythroblastic multinuclearity with positive acidified serum), accompanied with alpha thalassemia and secondary siderosis and diabetes.

Adult↗
PubMed1997

[Study on red cell enzymes and isoenzymes in patients with leukemia and myelodysplastic syndromes].

OBJECTIVE: To define the prevalence of acquired red cell enzymopathy in leukemia and MDS patients and explore its clinical significance. METHODS: Red cell enzymes (G6PD, 6PGD, PK, ENOL, ADA, PNP,ALD) activities and PK,ADA isoenzymes were assayed by the methods recommended by ICSH and PAGE electrophoresis. RESULTS: The prevalence rate of G6PD deficiency was 43.1%, and of PK deficiency was 27.8%, in leukemia and MDS patients. The prevalence rates of PK, G6PD and PNP deficiency in MDS patients were 50%, 48% and 39.1%, respectively, while of ADA, ENOL and ALD over production in MDS patients were 69. 6%, 40% and 30. 4%, respectively. PK activity was increased in CML, while the PK isoenzyme pattern was normal. Administration of antitumor antibiotics to AL decreased G6PD and 6PGD activities (P < 0.05). ADA2 was expressed in MNC of AML (M4 and M5) and in RBC of CMML, but not expressed in ALL (P < 0.001). CONCLUSION: Determination of enzymes may be used as prognostic parameters in CML,a predictor of antitumor drug resistance in acute leukemia, and a differential parameter between MDS-RA and aplastic anemia and between CML and CMML.

Adolescent↗