**2. Method**

The prospective study of laboratory was performed on 90 known patients with chronic kidney diseases( CKD) complicated with chronic renal failure(CRF), admitted in hospital, prior to undergoing schedules of dialysis, (55 men and 35 women), in average ages 35-65 years (mean, age 50, SD= +\_2).

The patients were analyzed once a month, all at the same day, to connection and after connection of hemodialysis schedules, in medical internal department.

For diagnosis of specific anemia of chronic renal diseases, laboratory tests included hemoglobin (HGB), hematocrit (HCT), white blood cells and platelets count, differential count and red cell indices (mean cell volume (MCV), mean cellular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) red cell distribution width (RDW), being performed using an automated analyzer (Coulter HMX with 22 parameters) and for

The MCV is an indicator of the size of red blood cells. MCV is measured in cubic

Mean corpuscular hemoglobin (MCH) measures the amount, or the mass, of hemoglobin present in one RBC. The weight of hemoglobin in an average cell is obtained by dividing the

(Reference values: adult men; MCH = 27 - 31 pg, women = 27-30 pg). Mean corpuscular hemoglobin concentration (MCHC) measures the proportion of each cell taken up by

The results are reported in percentages, reflecting the proportion of hemoglobin in the RBC. The hemoglobin is divided by the hematocrit and multiplied by 100 to obtain the MCHC.

RDW (red cell distribution width) reflects the size distribution of the erythrocyte population. The hematological instrument calculates it as a coefficient of variation (CV),

The aim of this study was to identify the values and changes of hematological and biochemical parameters in blood samples between two consecutive schedules of hemodialysis and assesses the effect of plasma osmolality on errors of platelets count, to the hospitalized patients admitted in hospital with diagnosis chronic renal diseases complicated

The prospective study of laboratory was performed on 90 known patients with chronic kidney diseases( CKD) complicated with chronic renal failure(CRF), admitted in hospital, prior to undergoing schedules of dialysis, (55 men and 35 women), in average ages 35-65

The patients were analyzed once a month, all at the same day, to connection and after

For diagnosis of specific anemia of chronic renal diseases, laboratory tests included hemoglobin (HGB), hematocrit (HCT), white blood cells and platelets count, differential count and red cell indices (mean cell volume (MCV), mean cellular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) red cell distribution width (RDW), being performed using an automated analyzer (Coulter HMX with 22 parameters) and for

connection of hemodialysis schedules, in medical internal department.

micrometers or fento-liters (Reference values: adult men: 80-94 fl, women: 81-99 fl).

MCH is expressed in picograms of hemoglobin per cell (pg/L, 1 pg = 10-12 g).

hemoglobin by the total RBC count.

(Reference values: adults: MCHC = 32- 36 %}

(Reference values: adults RDW = 11.5 - 15.5)

with chronic renal failure.

years (mean, age 50, SD= +\_2).

**2. Method** 

hemoglobin.

specific biochemical parameters in chronic renal failure as serum iron, total iron binding capacity, and index saturation transferrin ( IST), usually and specific biochemical tests:

Glucose, Urea nitrogen, Creatinine, Sodium, Potassium, E CO2, was used a dry chemistry analyzer Vitros 700(Ortho Diagnostics), Johnson \$ Johnson.

Reticulocyte count (RET %) was calculated after microscopic analysis of brilliant cresyl blue stained slides, (normal ranges adult: 0.5 - 1.5%). To evaluate rate of erythropoiesis, the Reticulocyte Production Index (RPI) was calculated using the formula: [RPI = RET% x HCT patient /45 /reticulocyte time maturation], where maturation time (reticulocytes survival days in peripheral blood**)** was considered 1 day

for HCT 36-45%, 1.5 days for HCT 26-35%, 2 days for HCT 16- 25% and 2.5 days for HCT < 15%. Reference interval for RPI in healthy individuals is 1.0-2.0; and RPI < 2 in a person with anemia indicates ineffective erythropoiesis, while values > 2 indicate compensation for decreased red cell survival (bleeding, hemolysis) [11].

Three methods were used to assess platelet counts of hemodialysis patients: optical microscopy, peripheral blood smear and user of the cytometry principle with impedance principle (VIC) by Coulter HNX hematological analysis. For to avoid systematic errors during platelets count by optical microscopy, a method of direct counting in the Burker-Turk chamber( hemacytometer) has been recommended for use in parallel with determination of the number of platelets counted on peripheral blood smear, ( by optical microscopy). Calculation of the platelets counted in the Burker-Turk chamber considers the height of the chamber and the surface of the middle square of the chamber to yield a value of 0.2mm². [12].

The calculation formula for hemacytometer cell counts determines the number of cells within 1μL (1 mm³) of blood. To make this determination, the total number of cells counted must be corrected for the initial dilution of blood and the volume of diluted blood used. The standard dilution of blood for platelet counts is 1:100; therefore the dilution factor is 100. The volume of diluted blood used is based on the area and depth of the counting area. The area counted is 2 mm² and the depth is 0.1 mm; therefore the volume factor is 0.2 mm³. Total number of cells counted • dilution factor • 1/volume factor = cells/mm3 (cells/mm³= cells/μL or cells/μL • 10³μL /L = cells x 109/L).

Direct microscopy of the blood smear yields the number of thrombocytes count by counting those found between 1000 erythrocytes (5 microscopic fields of 200 red cells) multiplied by the number of erythrocytes/mm.³ and then divided /1000) with the results expressed as platelets/ mm³. The estimate of platelet count from slides uses a semiquantitative method, whereby 1 platelet / oil immersion field is equivalent with 20.000 plt/mm³. Figure 1 In optical microscopy, one assesses a panoptic colored blood smear under the immersion objective(100 X). Most platelets have a dendritic aspect and fringe-like extension. Normal platelets have diameter of 2-4 microns on the blood smear with 70% alone, 20% in groups of 2 or 3 and 10% in larger groups or "big pools". Correctly executed blood smear reveal microscopic fields on the oil-immersion objective with an average of 10 platelets;as either isolated or grouped. Visualization of <5 platelets on the microscopic field connotes thrombocytopenia while >40 indicates thrombocythemia. [13].

Platelets are typically disk-shaped with a more dense central (granular) area and a peripheral (crystalline) area with functional dendritic fringes [14]. If activated by toxic metabolic factors, platelets become more spherical, which can yield a decrease in the intensity of the image in the microscopic lenses, due to light transmission and diffusion through samples. When platelets are activated, they become spherical with a hypogranular

Variability of Biological Parameters in Blood Samples

Between Two Consecutive Schedules of Hemodialysis 109

To the 54 patients with anemia of chronic kidney diseases (ACKD) and chronic renal failure( CRF) were registered in 30.90% of cases normal TIBC values (mean value 282 microgram/ d L, SD=2.5), low RPI in mean value of 1.33, low IST in mean value of 7.62%, with middle ineffective erythropoiesis and moderate iron deficiency anemia (IDA) and to 19.10 % of patients with ACKD and CRF associated with renal inflammations, were calculated low RPI, in mean value of 1.21, high TIBC value (mean value 468 microgram/d L, SD =2.4) and low

In biochemical field, in this study on this cohort of hemodialysis patients, was obtained the variability of plasma osmolality past normal individual values (310 Osm/l), in the samples taken from the patients with chronic renal failure because of high values of Urea nitrogen (mean value 112 mg%; 40 mmol/L; SD = 2.40); Creatinine (mean value 5.5 mg/%; 4.85 mmol/L); SD=0.15); Sodium (mean value 170 mmol/L; SD=0.14); Potassium (mean value 14.5 mmol/l; SD=2.88); E CO2 (mean value 11 mmol/L; SD=0.26). Prevalence of anemia to patients admitted in hospital for undergoing schedules of hemodialysiss have been registered in percents: 60% of cases, with normochromic-normocytic anemia, 30% of cases with microcytic-hypochomic anemia and nutritional iron deficiency, 7% of cases with

In cases with microcytic-hypochomic anemia and nutritional iron deficiency were registered by this study that mean corpuscular volume (MCV) of red cells decreases below normal value before that the hemoglobin to be decreased under normal value. Iron deficiency




In the two cases of study were registered suspect flags on Coulter HMX: neutropenia, lymphopenia and increased MCV erythrocyte index (109 f L). On blood smear from peripheral blood, in optic microscopy the reticulocyte count was decreased (0.4%), and neutrophil granulocytes showed multi-segmented nuclei, macrocytes (larger than normal RBCs) presence of ovalocytes (oval-shaped RBC) but Howell-Jolly bodies(chromosomal remnant) was absented. An elevated MCV should not be ignored because the patient is especially suspected of alcohol abuse. Blood chemistries will also showed: an increased lactic acid dehydrogenase

Bone marrow (checked in a patient suspected of megaloblastic anemia on hematological analyzer, in 3% from cases) showed megaloblastic hyperplasia~ 45%, ploycromathopil and acidophil erythroblasts with megaloblastic character, large metamielocytes and giant band forms. Biopsy results from gastric mucosa showed lesions of chronic gastritis, non-atrophic epithelium and the patient was receiving the recommendation from clinician doctor to

(LDH) values of .increased of homocysteine, folic and vitamin B12 deficiency.

IST in mean value of 6.5%, with severe ineffective erythropoiesis and severe IDA.

aplastic anemia and 3% with macrocytic and vitamin B12 deficiency.

erythrocyte, level of iron serum value, with high TIBC (8%),

anemia associated with ACKD was presented in three forms:

cells (10%),

IDA [19].

assess B12 vitamin.

cytoplasm and release small particles. This may lead to the erroneous detection of platelets when using the microscopy owing to their deformed morphology.

#### Fig. 1.

Recognizing erroneous results of platelet counts is especially critical for a consistent decision in the diagnosis of disseminated intravascular coagulation (DIC) and for clinical decision making regarding transfusion.

The platelet count is an indispensable parameter in the DIC scoring system proposed by the International Society on Thrombosis and Hemostasis Sub-Committee of the Scientific and Standardization Committee on DIC, in which platelet counts of less than 100 × 103/μL (100 × 109/L) and less than 50 × 103/μL (50 × 109/L) would score 1 and 2 points, respectively. [15, 16] The samples were assessed for platelet count by statistical parameters: [SD = (Xi- Xm )2 /n – 1; accuracy: (%Diff = X average – X target/ X mean x 100, with normal value until + − 25) and Z score( Z = X average-X target/SD, with normal value until +-2, R>0.95%), for average platelets 150-400 x10³/μl, 95% CI.].
