The Sysmex SE-9c500 is a new, fully automated haematology analyser, providing a complete blood count (CBC), including a five-part differential count (DC), with flagging of morphological abnormalities. The SE-9500 was evaluated according to guidelines published by the International Committee for Standardisation in Haematology (ICSH). The results demonstrated minimal carryover (< 0.01%) and excellent linearity for WBC, RBC, HGB and platelet (PLT) (r>> 0.995). Samples were stable with regard to CBC parameters after storage for up to 48 h at room temperature (RT) and 4 degrees C. Imprecision was generally acceptable for all CBC parameters (CV < 5%). Correlation between the SE-9500 and reference methods was excellent (r>> 0.97) for all the major CBC parameters (WBC, RBC, HGB, PLT). There was minimal interference for WBC, RBC, HGB and PLT at high concentrations of bilirubin (BIL=224 micromol/l) or triglyceride (TG=7.78 mmol/l). SE-9500 reference values for CBC parameters are presented. Our results indicate that the SE-9500 is an excellent tool for routine haematological examination.

26 patients, in whom hysterectomy and bilateral ovariectomy had been performed for a gynecological indication several years before, were tested for urinary testosterone (by a gas chromatographic method) and interstitial cell-stimulating hormone (ICSH) (by a hemagglutination-inhibition method). 12 patients did not develop any pathology of the breast 8.1 +/- 2.2 years after ovariectomy (control group), and 14 patients developed breast cancer 13.0 +/- 3.0 years after ovariectomy (breast cancer group). The average number of years following ovariectomy did not differ significantly between the two groups. There was no significant difference in the mean values of testosterone and of ICSH excretion between the patients of the control group and those of the breast cancer group, but a significant proportion (6 out of 14) of the breast cancer group patients had a much higher testosterone excretion than the highest level (8.4 microgram/24 h) in patients of the control group. This incidence (42.8%) tested by the chi-square method was found to be significant (p less than 0.02).

Schistocytosis is the morphological hallmark of the microangiopathic haemolytic anaemia of thrombotic microangiopathy (TMA). Consensus guidelines for manual schistocyte quantitation are available, but limited research has evaluated them. The 2012 International Council for Standardization in Haematology (ICSH) recommends a schistocyte quantitation of 1% as a robust cut-off for significance, with the quantitation including helmet, crescent, triangle and keratocyte poikilocytes; and microspherocytes only in the presence of helmets, crescents/triangles, and keratocytes. We aimed to evaluate the relative contribution of these different poikilocytes to schistocyte counting; compare the ICSH method with our proposed method which counts only cells most specific for red cell fragmentation (helmet, crescent and triangular schistocytes); and evaluate inter- and intra-observer agreement. Blood films were sourced from the Australian Snakebite Project, including non-envenomed and envenomed cases, with and without TMA. In blood films across the range of schistocytosis, the predominant poikilocytes present were helmets and crescents. Triangles, keratocytes and microspherocytes were typically only present when ICSH schistocyte count was >1%. With results dichotomised as <1.0% or ≥1.0%, our proposed new method versus the ICSH method showed almost perfect agreement [observed agreement 95%, Cohen's kappa (κ)=0.84, SE 0.04, 95% CI 0.76-0.92, p<0.005]. Inter-observer strength of agreement for our method was moderate (Fleiss' κ for comparisons between three non-unique microscopists κ=0.50, SE 0.05, 95% CI 0.41-0.59, p<0.005). Intra-observer reproducibility assessed in two microscopists ranged from substantial (Cohen's κ=0.71, SE 0.08, 95% CI 0.55-0.86, p<0.005) to borderline almost perfect agreement (Cohen's κ=0.81, SE 0.07, 95% CI 0.68-0.93, p<0.005). Schistocyte quantitation using our new method is simpler than the 2012 ICSH method and had almost perfect agreement. Our finding of moderate inter-observer agreement in quantitating helmet, triangle and crescent schistocytes is applicable to both the ICSH and our newly proposed method. This finding underscores the importance of clinicopathological correlation and repeated examinations in the context of a clinically suspected TMA.

Keywords: Schistocytes; erythrocytes; morphology; thrombotic microangiopathy.

OBJECTIVE

To study the endocrine disturbing effect of fluorin on Hypothalamus-Hypophysis-Testis axis in male rats.

METHODS

A total of 36 Wister male rats weighting 60-70 g were randomly divided into group I (high fluoride group of F-100 mg/l), group II (low fluoride group of F- 30 mg/l), group III (control group with pure water), with 12 rats in each group. Fluoride was administered with drinking water for 8 weeks. Then the level of procreation hormone in serum was detected by RIA method. And the spermatozoa quality was analysized.

RESULTS

There was difference between group I, group II and group III each other (P < 0.05) in body weight. As to right testis weight, there was difference between group I, group II and group III each other (P < 0.05). Epididymide organic coefficient in group II and group I were lower than that in group III (P>> 0.05). Compared with group III, the counts amount of sperm and the rates of sperm mobility in group II and group I singnificantly increased (P < 0.05), and the rates of sperm aberration in group II and group I significantly decreased (P < 0.05), compared with group II, the sperm quality of group I descreased significantly (P < 0.05). The level of GnRH in three groups were significant difference between each groups (P <0.05). The level of FSH in three groups were significant difference between each groups (P < 0.05). The level of ICSH in three groups were no significant difference between each groups (P>> 0.05). The level of T in Group I is significant lower than that of in Group II and Group III (P < 0.05). The level of E2 in Group I is significant higher than that of in Group II and Group III (P < 0.05).

CONCLUSIONS

These data support our hypothesis that fluorin have no significant effect on the function of male Rat Hypothalamus, that fluorin may effect the function of male Rat Hypophysis, that T/ICSH can be used to find the functional change of Leydig cells. Fluorin could affect hormone levels of each layer of the Hypothalamus-Hypophysis-Testis axis, and show the male reproductive endocrine disturbing effects.

Effect of soybean (Glycine max) on serum level of some sex hormones: testosterone, follicle stimulating hormone (FSH), luteinizing hormone/interstitial cell stimulating hormone (LH/ICSH), estradiol and prolactin in male rats was investigated. Twenty male albino rats of 12 weeks old with similar body weights were assigned to four groups of 5 rats each and treatment with soybean meal at 100, 200 and 300 mg kg(-1) body weight, respectively daily for 8 weeks. Blood samples collected through cardiac puncture were assayed for levels of hormones. There were dose-dependent effects of the soybean meal on the serum concentration of the hormones. The treatment significantly reduced the levels of testosterone and FSH in the serum while it significantly increased the levels of estradiol, LH/ICSH and prolactin. The results show that soybean (Glycine max) had strong capability to disrupt hormonal functions. Hence, its indiscriminate use could increase the risk of infertility in males.

The function of the hypothalamo-pituitary-gonadal system has been studied in 118 males with different stages of alcoholism. Comparison with the control group has shown decreased levels of interstitial cell stimulating hormone (ICSH), follicle-stimulating hormone (FSH), and testosterone, and elevated concentrations of prolactin (Prl). Differences in the parameters of hormonal activity expressed in a varying degree of a decrease in FSH, ICSH, and testosterone, and a Prl elevation are related to change in mutual regulatory influences between the gonads and hypothalamo-pituitary complex. The most informative parameters were concentrations of Prl and testosterone (considering elevation in the former and reduction in the latter hormone) and the ratio of their concentrations. Difference of these parameters in each group (corresponding to the stages of alcoholism) may be used as an additional diagnostic criterion of chronic alcohol intoxication.

A semiautomatic electronic blood cell counter (Sysmex F-800) was evaluated with equine blood, according to the protocol of the International Committee for Standardization in Haematology (ICSH, 1984). The precision and overall reproducibility were acceptable for all the parameters studied except for the platelet count, in which a coefficient of variation of 18.8% and 21.7% was obtained for within and between batch precision and 26.76% for overall reproducibility. Carry-over for the haematocrit value and platelet count was unsatisfactory, thus the use of a blank diluent sample between different analyses was necessary. Linearity of the analyser was acceptable in the range studied. A total of 100 equine blood samples were studied with the Sysmex F-800 and the manual method. The correlations between both Methods were acceptable for all the parameters. In the storage study, the red blood cell count and the haemoglobin concentration were the parameters with longest stability (72 h) at 4 degrees C and 25 degrees C. Platelet count decreased significantly at 3 h post-extraction in those samples stored at 4 degrees C. During the evaluation period, Sysmex F-800 was easy to handle and it is well suited for routine equine blood cell analysis.

OBJECTIVE

This was a preliminary study on the prevalence of the HbS gene, associated with sickle cell disease, other hemoglobinopathies and G6PD deficiency of immigrant and non-immigrant population of Catalonia.

METHODS

A total of 3,189 blood samples from the Catalan Neonatal Screening Program for Metabolic Diseases (CNSPMD) including 1,620 from immigrant population were screened for haemoglobinopathies and G6PD deficiency. For screening of hemoglobinopathies the high performance liquid chromatography (HPLC) method was used and for the screening of G6PD deficiency, we used the fluorescent spot test as described by ICSH.

RESULTS

1. Hemoglobinopathies: in 47 samples from immigrant population 2 cases of sickle cell anemia (phenotypes FS and FSC) were detected as well as 45 cases of heterozygote carriers of different pathological hemoglobins (HbS, HbC, HbD and HbE). 2. G6PD deficiency: in 29 samples, 3 cases of G6PD deficiency belonging to local (non-immigrant) population of G6PD were detected.

CONCLUSIONS

The incidence of sickle cell disease in the risk population of Catalonia is 1 case out of 810 samples. This value is significantly higher than that reported for any of the metabolic diseases included in the CNSPMD. Despite it is a preliminary study, the results obtained give further support to the convenience of incorporating a neonatal screening of hemoglobinopathies, at least in the risk population, to the official programs of newborn screening. Due to its feasibility and low cost, a similar criterion might be adopted for the neonatal screening of G6PD deficiency.

The effect of a low-dose treatment (10 mg/day) of stanozolol on the anaerobic threshold (AT) and on maximal velocity (Vmax) was studied in ten well-trained runners in a 6-week trial experiment. The serum levels of testosterone (T), interstitial cell-stimulating hormone (ICSH), and of follicle-stimulating hormone (FSH) were determined before, during, and after the steroid administration. No improvements of AT and Vmax were found. No improvement of the competition running times was observed during the experimental period. The ICSH and FSH serum concentrations did not change significantly, while a marked decrease of the T serum concentration was observed. It is concluded that a low-dose treatment of stanozolol, sufficient to depress the testicular function, does not improve athletic performance.

The purpose of the present study was to find out whether there is a correlation between the urinary levels of testosterone or of estrogens (estrone and estradiol) and ICSH urinary concentration in normal postmenopausal women. Examination of 35 normal subjects at different postmenopausal ages (from one year to over 10 years after menopause) showed a highly significant inverse correlation between ICSH and testosterone urinary excretion levels (coefficient of correlation "r" equals minus 0.548), but none between the urinary estrogen excretion levels and ICSH urinary concentrations.

Field investigation on labour hygiene in two plants producing trinitrotoluene (TNT) in Henan Province showed most air TNT levels in the workplace exceeded national maximum allowable concentration (MAC 1 mg/m3) and the skin of the workers exposed to TNT was severely contaminated. Determinations of serum levels of sex hormones showed those of testosterone were lower, but those of interstitial cell-stimulating hormone (ICSH or LH) and follicle stimulating hormone (FSH) were higher in workers exposed to TNT than that in controls with statistical significance (P < 0.01).

Mithun is a domesticated free-range bovine species primarily used as a meat animal and is a pride of North Eastern Hilly regions of India. The present study was conducted to measure the effect of seasons on scrotal & testicular biometrics and endocrinological profiles for different age groups at different seasons in mithun bulls. A total of 30 mithun males were selected from the mithun breeding farm, ICAR-NRC on Mithun, Medziphema, Nagaland, India and were equally distributed into five groups based on their age. Each group consisted of six animals and the groups were Gr A (0.1-1.0 year), Gr B (1.1-2.0 years), Gr C (2.1-3.0 years), Gr D (3.1-5.0 years) and Gr E (5.1-6.0 years). The seasons were grouped into winter, spring, summer and autumn based on the meteorological data such as temperature humidity index (THI) and sunshine hours. Scrotal circumference (SC) & testicular biometrics and endocrinological profiles such as follicle stimulating hormone (FSH), luteinizing hormone/interstitial cell stimulating hormone (LH/ICSH), testosterone, thyroxine (T4), cortisol and insulin like growth factor 1 (IGF 1) were estimated during different seasons for different age groups. Statistical results revealed that the SC & testicular biometrics and endocrinological profiles differed significantly (p < 0.05) among the different age groups for the same season whereas SC and endocrinological profiles significantly (p < 0.05) differed among the seasons for same age group. Significantly (p < 0.05) greater SC and testicular weight were observed in Gr E and D, lower in Gr A and B and intermediate in Gr C and increased as age advances. Significantly (p < 0.05) greater SC was observed in winter and spring and lowest was in summer season. The hormone profiles such as FSH, LH/ICSH, testosterone & thyroxine were significantly (p < 0.05) greater and IGF-1 & cortisol were significantly (p < 0.05) lower in spring & winter than in summer season. The hormones, FSH, LH/ICSH and thyroxine increased significantly in Gr A followed by reduced in Gr B and then increased to Gr D and E, whereas concentration of testosterone, cortisol and IGF-1 increased according to age advanced. It was concluded that the spring and winter seasons has significantly greater beneficial effects than summer season on reproduction and artificial breeding programme in semi-intensive management of mithun in the present location.

The ESrT-system 200 comprises a 215 mm long vacuum aspiration venepuncture tube which contains anticoagulant diluent for the measurement of the erythrocyte sedimentation rate (ESR) without direct handling of the blood sample. This combines the advantage of a tube of "Westergren" length with a reduction in biohazard risk. Blood from 160 patients (ESR range 2-135 mm/first hour) was tested in parallel with the selected Westergren ESR method of the International Committee for Standardization in Haematology (ICSH) and a close correlation (r = 0.967) between the two methods was obtained. A second Westergren ESR, using anticoagulated but undiluted blood, was measured on 58 specimens to give an ICSH "expected" ESR. The ESrT-system 200 result was within 12 mm/first hour of the "expected" result for 91% of the specimens. This new ESR system is simple to use, does not require mathematical correction of the ESR reading for tube length, and gives results that are comparable with those obtained with the ICSH Westergren method.

Recently, recommendations for the production of thromboplastins have been issued by International Committees (ICSH/ICTH) to achieve a standardization of prothrombin time assays. Following such recommendations we have standardized two new PT reagents based on coagulometric and photometric methods which rely on the same sensitive human placental thromboplastin. Both PT reagents have been optimized especially for use in oral anticoagulant therapy. They offer high sensitivity and reproducibility, accurate lot standardization and therefore internationally comparable values by the expression of the ISI value of each reagent batch.