scholarly journals Frequency of Lentz Bodies Inclusion in Whole Blood Erythrocytes and Expanded Buffy Coat Smears

2019 ◽  
Vol 47 (1) ◽  
Author(s):  
Monica Alejandra Camargo Castillo ◽  
Bruno Albuquerque De Almeida ◽  
Felipe Yuji Okano ◽  
Angelica Menin ◽  
Stella De Feira Valle
Keyword(s):  
Blood Sample ◽  
Whole Blood ◽  
Blood Cells ◽  
Inclusion Bodies ◽  
Blood Smear ◽  
Buffy Coat ◽  
Median Frequency ◽  
Canine Distemper ◽  
Blood Smears ◽  
Significant Difference

Background: Canine distemper has been classified as highly contagious for most of domestic and wild carnivores, and the infection can be fatal. Canine distemper inclusion bodies, also denominated Lenz inclusion bodies, are large aggregates of viral nucleocapsid particles that can be form in red blood cells (RBCs), white blood cells (WBCs) and epithelial cells in many tissues during the acute phase of infection. Their presence in blood is transient and rarely encountered in light microscopy but are pathognomonic when identified in blood smears. The objective of this study was to investigate the frequency of distemper inclusions in erythrocytes according to the fraction of the sample used for blood smears. Materials, Methods & Results: The study was conducted with routine blood sample provided by the Veterinary Laboratory of Clinical Analysis from the Veterinary Teaching Hospital of Universidade Federal do Rio Grande do Sul. The EDTA-K2 blood sample of a 40 days old male dog, mixed breed, no immunization records, presenting diarrhea, hyporexia, myoclonus and pustules in the abdomen, was selected. In a routine peripheral blood smear examination, several distemper inclusions were observed in the erythrocytes. From this sample, ten smears were performed using a whole blood (WB) and top erythrocyte fraction combined with buffy coat, denominated of expanded buffy coat (EBC). The EBC fraction was obtained after centrifugation of EDTA whole blood in microhematocrit tubes at 9600 x g for 5 min to obtained the packed cell volume (PCV) and buffy coat. After centrifugation, the blood cells are separated into three layers based on density: platelets (adjacent to supernatant), WBCs, and RBCs in the bottom. The PCV was measured and the microhematocrit tube was ruptured 2% below the interface between leukocytes and plasma, deposited into a plastic microtubes, homogenized and used for blood smear preparation. All smears were stained with Diff-Quick Stain. The frequency of observation of RBCs with distemper inclusions bodies was performed under optical microscopy, in the immersion objective (100x), accounting for complete fields up to a minimum of 1000 RBCs, and compared between WB and EBC. In comparison between blood smears obtained from WB and EBC, a highly significant difference (P = 0.0004) was observed in the frequency distribution of distemper inclusion. The median of frequency of RBCs with distemper inclusions in a WB smears was 12.68/1000 RBCs (10.1 - 16.1/1000 RBCs), with a coefficient of variation (CV) of 12%. Median of frequency of distemper inclusions from EBC smears was 54.23/1000 RBCs (45-77.9/1000 RBCs), CV of 18% were observed. The median frequency of inclusions found in EBC smears was 4.27 times higher than the WB smears. Discussion: Buffy coat smear providing a concentrated preparation of nucleated cells and this procedure is useful to looking for low-incidence infectious organisms or other hematologic alterations. The upper fraction of the RBC column, below the buffy coat, is composed of young RBCs. Selection of these portion, and their possible formed in the bone marrow viral replication phase, could justified the increase in the frequency of RBCs containing viral inclusions in EBC, which would also increase the sensitivity of the technique. EBC was homogenized previously to make the smears, certifying the adequate cell distribution in the slide surface without interfere with the frequency of distemper inclusion in RBCs observation. These results were confirmed with the coefficients of variation. In conclusion, distemper inclusions bodies in RBCs from EBC is a recommended diagnosis method in patients suspected of canine Distemper infection. The observation being more frequent in the EBC in comparison with WB, commonly used in veterinary hematology.

scholarly journals Circulating and bone marrow myeloid cells containingLeishmaniaamastigotes in a case of advanced canine leishmaniosis

2019 ◽  
Vol 31 (5) ◽  
pp. 726-731
Author(s):  
Ioannis L. Oikonomidis ◽  
Theodora K. Tsouloufi ◽  
Mathios E. Mylonakis ◽  
Dimitra Psalla ◽  
Nectarios Soubasis ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Weight Loss ◽  
Blood Smear ◽  
Bone Marrow Aspiration ◽  
Buffy Coat ◽  
Routine Practice ◽  
Canine Leishmaniosis ◽  
Aspiration Cytology ◽  
Blood Smears ◽  
Leishmania Amastigotes

A 5-y-old male Poodle mix was presented with intermittent vomiting, anorexia, and weight loss. Physical examination revealed emaciation, lethargy, dehydration, hypothermia, respiratory distress, and splenomegaly. Based on clinicopathologic, serologic, and parasitologic findings, diagnoses of severe leishmaniosis and dirofilariasis were made. Extracellular, intraneutrophilic, and intramonocytic Leishmania amastigotes were observed on blood smear and buffy coat smear examination. In blood smears, 0.2% of neutrophils were observed to be infected; in buffy coat smears, 0.5% of neutrophils and 0.1% of monocytes were found to be infected. Leishmania amastigotes were also found engulfed by eosinophils and neutrophil precursors in bone marrow aspiration cytology. The detection of Leishmania amastigotes in blood smears is rare, and the clinical significance is uncertain. In circulating blood, Leishmania amastigotes are primarily found phagocytized by neutrophils. Although debatable, there is growing evidence that neutrophils are used as carriers enabling the “silent entry” of the protozoa into macrophages (“Trojan horse” theory). To date, cytologic screening of blood smears for the diagnosis of canine leishmaniosis is not a routine practice. Clinical pathologists and practitioners should be aware that Leishmania amastigotes may be present in neutrophils and less frequently monocytes during blood smear evaluation; neutrophil precursors and eosinophils may also be parasitized in bone marrow specimens.

scholarly journals Diabetes affects endothelial cell function and alters fibrin clot formation in a microvascular flow model: A pilot study

2020 ◽  
Vol 17 (1) ◽  
pp. 147916412090304
Author(s):  
Lorenz Jenny ◽  
Andreas Melmer ◽  
Markus Laimer ◽  
Elaissa T Hardy ◽  
Wilbur A Lam ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Whole Blood ◽  
Blood Cells ◽  
Plasma Proteins ◽  
Flow Model ◽  
Clot Formation ◽  
Microvascular Flow ◽  
Complex Interactions ◽  
Significant Difference

Diabetes is a proinflammatory and prothrombotic condition that increases the risk of vascular complications. The aim of this study was to develop a diabetic microvascular flow model that allows to study the complex interactions between endothelial cells, blood cells and plasma proteins and their effects on clot formation. Primary human cardiac microvascular endothelial cells from donors without diabetes or donors with diabetes (type 1 or type 2) were grown in a microfluidic chip, perfused with non-diabetic or diabetic whole blood, and clot formation was assessed by measuring fibrin deposition in real time by confocal microscopy. Clot formation in non-diabetic whole blood was significantly increased in the presence of endothelial cells from donors with type 2 diabetes compared with cells from donors without diabetes. There was no significant difference in clot formation between non-diabetic and diabetic whole blood. We present for the first time a diabetic microvascular flow model as a new tool to study clot formation as a result of the complex interactions between endothelial cells, blood cells and plasma proteins in a diabetes setting. We show that endothelial cells affect clot formation in whole blood, attributing an important role to the endothelium in the development of atherothrombotic complications.

scholarly journals Glucose transporter-4 in white blood cells of young and old sled dogs: a model for human biomarker development

Polar Record ◽  
2013 ◽  
Vol 51 (2) ◽  
pp. 160-164 ◽  
Author(s):  
Theresia M. Schnurr ◽  
Arleigh J. Reynolds ◽  
Lawrence K. Duffy ◽  
Kriya L. Dunlap
Keyword(s):  
Insulin Resistance ◽  
Blood Cells ◽  
Molecular Mechanisms ◽  
Glucose Transporter ◽  
White Blood Cells ◽  
Insulin Response ◽  
Buffy Coat ◽  
Insulin Concentration ◽  
Sled Dogs ◽  
Significant Difference

ABSTRACTThe insulin responsive glucose transporter, GLUT4 is found predominantly in muscle and adipose cells. Maratou and others (2007) reported that there is GLUT4 in white blood cells (WBC) collected from human subjects in response to insulin activation. This study was designed to validate the presence of GLUT4 in white blood cells of sled dogs and furthermore to investigate whether changes in levels of the GLUT4 protein might be associated with aging. Additionally, we examined the blood insulin concentration of two populations of dogs, young and old, before and after a meal to observe their insulin response. It is documented in skeletal muscle that GLUT4 expression is increased as a result of conditioning, making sled dogs an excellent model in the circumpolar north for studying the effects of exercise, nutrition and diabetes (Felsburg 2002; Kararli 2006). Blood was withdrawn from 11 healthy sled dogs: 6 young (1–5 years) and physically fit, conditioned for racing and 5 old (7–13 years), retired from racing. The insulin response was determined using blood plasma and ELISA. The buffy coat (containing WBC) was collected with a glass pipette after centrifugation and washed and suspended in 1x phosphate buffer. GLUT4 was measured using ELISA kits (USCN Life Sciences). The results validate that GLUT4 is present in white blood cells in sled dogs. Age had no significant effect in the concentration of GLUT4 between the populations of old and young dogs. A significant difference in insulin levels pre and post meal in young (0.13 ± 0.03 ng/mL (pre), 0.22 ± 0.04 ng/mL (post), p < 0.05) and old (0.13 ± 0.02 ng/mL (pre), 0.22 ± 0.03 ng/mL (post), p < 0.05) dogs was observed, displaying the typical postprandial insulin spike. No significant difference was found in insulin concentration comparing old versus young dogs. Our data shows that white blood cells in young (40.4 ± 2.4 ng/mL) and old (35.3 ± 8.8 ng/mL) sled dogs have quantifiable but non-significant different GLUT4 levels (p > 0.05). Detecting GLUT4 via an ELISA in white blood cells, opens up minimally invasive avenues for studying the underlying molecular mechanisms associated with insulin resistance in more complex, dynamic and physiological systems. This project was the first step in developing a protocol for this simple, technique with a potential clinical application for diagnosing insulin resistance.

SEPARATING PLASMA AND BLOOD CELLS BY DIELECTROPHORESIS IN MICROFLUIDIC CHIPS

2012 ◽  
Vol 19 ◽  
pp. 185-189 ◽  
Author(s):  
TZONG-SHYNG LEU ◽  
ZHI-FENG LIAO
Keyword(s):  
Blood Sample ◽  
Flow Rate ◽  
Whole Blood ◽  
Blood Cells ◽  
Volume Fraction ◽  
Side Wall ◽  
Inlet Flow ◽  
Inlet Flow Rate ◽  
Side Walls ◽  
Whole Blood Sample

In this paper, a dielectrophoretic (DEP) micro separator is studied for plasma-blood separation. DEP forces created by non-uniform electric fields are used as deflected forces to deplete blood cells from side walls at a given inlet flow rate ( Qin ). Then one can extract plasma through a microchannel on side wall at certain extraction flow rate ( Qp ). In this experiment, saline isotonic solution is chosen as dilute solution for whole blood. The minimum dilute ratio (whole blood: saline dilute) is found to be 1:3 for DEP to substantially deplete blood cells from side walls. Exraction of plasma from whole blood sample by DEP force is also investigated. Experimental results show blood cells do not enter side channel by DEP force at inlet flow rate Qin =0.5 μ1/ min when plasma extract flow rates is Qp ≤ 0.3 μ1/ min . By calculating pure plasma extraction volume fraction, the efficiency in current experiment can reach as high as 20% if dilute ratio 1:3 of whole blood sample is considered.

Comparative methodology for the detection and differentiation of circulating microfilariae of Dirofilaria immitis in the dog

2004 ◽  
Vol 78 (2) ◽  
pp. 137-140 ◽  
Author(s):  
M.E. Mylonakis ◽  
E. Papadopoulos ◽  
A.F. Koutinas ◽  
C. Paitaki ◽  
L. Leontides
Keyword(s):  
Whole Blood ◽  
Dirofilaria Immitis ◽  
Blood Smear ◽  
Capillary Tube ◽  
Buffy Coat ◽  
The Other ◽  
Blood Samples ◽  
Comparative Methodology ◽  
Whole Blood Samples ◽  
Knott’S Test

AbstractThe sensitivities of the Knott's test (four 20-μl sediment aliquots), quantitative buffy coat capillary tube method (QBC tube, 111 μl of whole blood) and direct blood smear (DBS, 20 μl of whole blood) were evaluated for the detection of microfilaraemia in dogs. Undiluted whole blood samples taken from 70 Dirofilaria immitis antigen-positive dogs and 10 serially diluted microfilaraemic blood samples at concentrations of 400, 200, 100, 50, 25 and 12 microfilariae (mff) ml−1 were examined. For filarial speciation, the buffy coat of QBC tubes was mixed with one drop of methylene blue–formalin solution and examined as a direct smear. In 52/70 microfilaraemic blood samples, the number of mff ranged from 12 to 321987 ml−1 (median: 3199 ml−1). The diagnostic sensitivity of the Knott's test, QBC tube method and DBS in undiluted blood samples attained the 100%, 98% and 92.3% levels, respectively. Eighteen dogs tested amicrofilaraemic by all three methods. At concentrations of 400 mff ml−1, a 100% sensitivity was found by all three methods, while at 200 mff ml−1 the Knott's test, QBC tube and DBS were 100%, 100% and 90% sensitive, respectively. The relevant figures at 100 mff ml−1 were 100%, 100% and 80%, at 50 mff ml−1 100%, 100% and 50%, at 25 mff ml−1 100%, 100% and 10% and at 12 mff ml−1 80%, 50% and 10%. At 50 and 25 mff ml−1, the DBS was less sensitive compared to the other two methods, while at 12 mff ml−1, only to the Knott's test. A significant correlation was found between the QBC tube method and Knott's test regarding mff speciation. Therefore, the QBC method may be considered a reliable alternative to the Knott's test for both the detection and speciation of mff in the dog.

scholarly journals Detection of the Plasmodium falciparumAntigen Histidine-Rich Protein 2 in Blood of Pregnant Women: Implications for Diagnosing Placental Malaria

1999 ◽  
Vol 37 (9) ◽  
pp. 2992-2996 ◽  
Author(s):  
Rose F. G. Leke ◽  
Rosine R. Djokam ◽  
Robinson Mbu ◽  
Robert J. Leke ◽  
Josephine Fogako ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Pregnant Women ◽  
Whole Blood ◽  
Peripheral Blood ◽  
Blood Smear ◽  
Placental Malaria ◽  
Malarial Infection ◽  
Peripheral Plasma ◽  
Blood Smears ◽  
Peripheral Blood Smears

Pregnant women have an increased susceptibility to infection byPlasmodium falciparum. Parasites may be present in the placenta yet not detectable in peripheral blood smears by routine light microscopy. In order to determine how frequently misdiagnosis occurs, peripheral blood and placental samples were collected from 1,077 Cameroonian women at the time of giving birth and examined for the presence of malarial parasites by using light microscopy. Results showed that 20.1% of the women who had placental malaria were peripheral blood smear negative. Thus, malarial infection was not detected by microscopic examination of peripheral blood smears from approximately one out of five malaria-infected women. Since P. falciparum parasites secrete histidine-rich protein 2 (HRP-2), we sought to determine if detecting HRP-2 in either peripheral plasma or whole blood might be used to diagnose the presence of parasites “hidden” in the placenta. Samples of peripheral plasma from 127 women with different levels of placental malarial infection were assayed by HRP-2-specific enzyme-linked immunosorbent assay. HRP-2 was detected in 88% of the women with placental malaria who tested negative by blood smear. Additionally, whole blood was obtained from 181 women and tested for HRP-2 with a rapid, chromatographic strip test (ICT). The ICT test accurately detected malarial infection in 89.1% ofP. falciparum-infected women. Furthermore, 94% of women with malaria were accurately diagnosed by using a combination of microscopy and the ICT test. Thus, detection of HRP-2 in conjunction with microscopy should improve diagnosis of malaria in pregnant women.

scholarly journals Detection of Sickle Cell Anemia Through Contour Evidence Extraction and Estimation

2021 ◽  
Vol 10 (6) ◽  
pp. 182-191
Author(s):  
Aruna N S. ◽  
◽  
Dr. Hariharan S. ◽  
Keyword(s):  
Blood Sample ◽  
Sickle Cell ◽  
Sickle Cell Anemia ◽  
Blood Cells ◽  
Blood Smear ◽  
Automated System ◽  
Contour Extraction ◽  
Sickle Cells ◽  
Seed Points ◽  
Blood Smear Image

Diagnosis of sickle cell anemia by manual visual inspection through microscope is time consuming and causes human errors. Observational errors occur mostly due to overlapping of cells in blood smear image. Here, an automatic segmentation approach is introduced which isolates sickle cells from all other cells within a blood sample. The proposed system is an approach to find the elliptically shaped sickle cells through geometric feature extraction and contour based segmentation to isolate sickle cells. This technique is a method of isolating sickle cells from other cells within blood sample using cell morphology. A combined approach of extraction of seed points, contour extraction and estimation of contours is used for separation of sickle cells from red blood cells. The methods used for the extraction of seed points are by Ultimate Erosion for Convex Sets and Fast Radial Symmetry transform. The contour evidence is extracted by associating edges of the cells to the seed points. The overlapping and clustered cells in image are identified using ellipse fitting method for contour estimation. Using the seed points and the contour extraction, the edges of the cells are estimated. The lines joining the shape of cells are drawn through estimation of shape of contour. This eliminates cells other than elliptical shaped cells. The proposed system can successfully isolate sickle cells from healthy blood cells within the blood smear image. This automated system has a better accuracy and faster computation speed compared to the existing methods for the detection of sickle cells. This identification methodology helps the health professionals for faster diagnosis.

Manganese content of the cellular components of blood

1990 ◽  
Vol 36 (3) ◽  
pp. 450-452 ◽  
Author(s):  
D B Milne ◽  
R L Sims ◽  
N V Ralston
Keyword(s):  
Whole Blood ◽  
Blood Cells ◽  
Graphite Furnace ◽  
Manganese Content ◽  
Atomic Absorption Spectrophotometry ◽  
Buffy Coat ◽  
Absorption Spectrophotometry ◽  
Graphite Furnace Atomic Absorption ◽  
Turn Over ◽  
Cellular Components

Abstract We measured the manganese content of whole blood, plasma, platelets, mononucleated cells, polymorphonucleated cells, and erythrocytes. Platelets and blood cells were separated from whole blood by use of discontinuous gradients of colloidal polyvinylpyrrolidone-coated silica (Percoll), and their manganese content was measured by Zeeman graphite furnace atomic absorption spectrophotometry, after digestion with nitric acid and hydrogen peroxide. Erythrocytes account for about 66% of the manganese in whole blood, whereas the "buffy coat"--platelets and leukocytes--accounts for about 30%. The "buffy coat" components turn over more rapidly than do erythrocytes, so their manganese content may better indicate the body's manganese status.

scholarly journals Process Improvement by Eliminating Mixing of Whole Blood Units after an Overnight Hold Prior to Component Production Using the Buffy Coat Method

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Cherie Mastronardi ◽  
Peter Schubert ◽  
Elena Levin ◽  
Varsha Bhakta ◽  
Qi-Long Yi ◽  
...  
Keyword(s):  
Quality Control ◽  
Whole Blood ◽  
Buffy Coat ◽  
Activity Levels ◽  
Significant Difference ◽  
Quality Control Criteria ◽  
Centrifugation Step ◽  
Control Criteria ◽  
Quality Assessments ◽  
Component Production

The elimination of a thorough manual mixing of whole blood (WB) which takes place following the overnight hold, but before the first centrifugation step, during buffy coat component production at Canadian Blood Services (CBS) was investigated. WB was pooled after donation and split. Pairs of platelet, red blood cell (RBC), and plasma components were produced, with half using the standard method and half using a method in which the mixing step was eliminated. Quality assessments included yield, pH, CD62P expression and morphology for platelets, hemoglobin, hematocrit, hemolysis, and supernatant K+ for RBCs, and volume and factor VIII activity levels for plasma. All components, produced using either method, met CBS quality control criteria. There were no significant differences in platelet yield between components produced with and without mixing. A significant difference was seen for RBC hemolysis at expiry (P=0.03), but for both groups, levels met quality control requirements. Noninferiority of components produced without mixing was confirmed for all parameters. Manual mixing is laborious and has a risk of repetitive strain for production staff and its significance is unclear. Elimination of this step will improve process efficiencies without compromising quality.

WT-MO Algorithm

2019 ◽  
pp. 39-79
Author(s):  
Ana Carolina Borges Monteiro ◽  
Yuzo Iano ◽  
Reinaldo Padilha França ◽  
Navid Razmjooy
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Blood Smear ◽  
White Blood Cells ◽  
Morphological Operations ◽  
Visual Examination ◽  
Blood Smears ◽  
Computer Aided ◽  
The Future ◽  
Medical Analysis

Visual examination of blood smears is an essential tool for analysis, prevention, and remediation of several types of maladies. The interest of computer-aided decision has been acknowledged in many medicinal instances (e.g., automatic ways and means are being explored to spot, classify, and measure visual items in hematological cytology [HC]). This chapter proposes an entirely automated blood smear diagnosis system for hemograms, which can lessen the time spent to scrutinize a slide. The present framework relies on morphological operations (MOs) and soft segmentation by means of the watershed transform (WT). Experiments demonstrate the method efficacy to count white blood cells (WBCs) and red blood cells (RBCs). Some considerations about implementations, design advice and possible variants, as well as improvements are discussed. The future of automated medical analysis is contemplated.

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