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Acta Cytologica 2017Cytodiagnoses of specific malignancies are enabled through analyses of abnormal nuclear chromatin and cytoplasmic features in stained cells. (Review)
Review
OBJECTIVE
Cytodiagnoses of specific malignancies are enabled through analyses of abnormal nuclear chromatin and cytoplasmic features in stained cells.
AIM
The objective of this work was to explore the inception, development, and chemistry of the Pap stain method introduced in 1942 by Dr. G.N. Papanicolaou.
STUDY DESIGN
To achieve this, we carried out a review of the English literature.
RESULTS
Between 1914 and 1933, Papanicolaou first analyzed vaginal squamous cells in guinea pigs and later in human vaginal fluid samples using hematoxylin and eosin with limited color reactions, correlating the cell-type morphology with endocrinology and histology. The 5-dye Pap stain method evolved through 2 salient phases. The first, between 1933 and 1942, saw the introduction of alcohol-ether fixation and aqueous waterblue staining to enhance cellular transparency, aiding the distinction of cervical cancer cells from benign cells, with quantitative and qualitative assessment of squamous cell maturity. The second phase, between 1942 and 1960, saw the introduction and refinement of various alcoholic cytoplasmic counterstaining schemes with orange G and EA (light green, Bismarck brown, eosin) and phosphotungstic acid, allowing wider ranges of polychromasia and further enhancing cellular visualization, facilitating the distinction of cell types and improving diagnostic confidence.
CONCLUSIONS
Development of the Pap stain method followed specific historical and scientific events. The staining method evolved following incremental improvements in cellular transparency achieved through tailored cellular fixation and cytoplasmic staining using variable dye and pH combinations.
Topics: Animals; Coloring Agents; Cytoplasm; Female; Humans; Papanicolaou Test; Staining and Labeling; Uterine Cervical Neoplasms; Vaginal Smears
PubMed: 28384641
DOI: 10.1159/000457827 -
Comparative Medicine Apr 2015Hematologic parameters are important markers of disease in human and veterinary medicine. Biomedical research has benefited from mouse models that recapitulate such... (Comparative Study)
Comparative Study Review
Hematologic parameters are important markers of disease in human and veterinary medicine. Biomedical research has benefited from mouse models that recapitulate such disease, thus expanding knowledge of pathogenetic mechanisms and investigative therapies that translate across species. Mice in health have many notable hematologic differences from humans and other veterinary species, including smaller erythrocytes, higher percentage of circulating reticulocytes or polychromasia, lower peripheral blood neutrophil and higher peripheral blood and bone marrow lymphocyte percentages, variable leukocyte morphologies, physiologic splenic hematopoiesis and iron storage, and more numerous and shorter-lived erythrocytes and platelets. For accurate and complete hematologic analyses of disease and response to investigative therapeutic interventions, these differences and the unique features of murine hematopathology must be understood. Here we review murine hematology and hematopathology for practical application to translational investigation.
Topics: Animals; Bone Marrow; Disease Models, Animal; Hematologic Diseases; Hematology; Hematopoiesis; Humans; Mice; Pathology, Veterinary; Translational Research, Biomedical
PubMed: 25926395
DOI: No ID Found -
Journal of the American Veterinary... Jun 2011To determine diagnostic accuracy of using erythrocyte indices and polychromasia to identify regenerative anemia in dogs.
OBJECTIVE
To determine diagnostic accuracy of using erythrocyte indices and polychromasia to identify regenerative anemia in dogs.
DESIGN
Retrospective and prospective cross-sectional study.
ANIMALS
4,521 anemic dogs.
PROCEDURES
CBC results obtained between July 2002 and July 2008 by use of an automated laser-based flow cytometric hematology analyzer from dogs with Hct values ≤ 35% were retrieved. Sensitivity, specificity, accuracy, and predictive values of using erythrocyte indices and polychromasia to identify regeneration were determined, with a reticulocyte count > 65,000 reticulocytes/μL considered the gold standard. Similarly, 134 blood samples from anemic dogs were analyzed prospectively with an in-house electrical impedance analyzer.
RESULTS
Of 4,387 dogs with samples analyzed retrospectively, 1,426 (32.5%) had regenerative anemia. Of these, 168 (11.8%) had macrocytic hypochromic anemia. High mean cell volume and low mean cell hemoglobin concentration had low sensitivity (11%), high specificity (98%), and moderate accuracy (70%) when used to identify regenerative anemia. Use of polychromasia alone had an accuracy of 77%, and use of polychromasia combined with a high RBC distribution width (RDW) had an accuracy of 79%. Results obtained with the in-house analyzer were similar.
CONCLUSIONS AND CLINICAL RELEVANCE
Results suggested that most regenerative anemias in dogs were not macrocytic hypochromic. Polychromasia, with or without high RDW, was a more accurate indicator than other erythrocyte indices of regenerative anemia. To avoid a false diagnosis of nonregenerative anemia, a blood smear should be evaluated in anemic dogs when a reticulocyte count is not available.
Topics: Anemia; Animals; Blood Cell Count; Cross-Sectional Studies; Diagnosis, Differential; Dog Diseases; Dogs; Erythrocyte Count; Erythrocyte Indices; Female; Hematocrit; Hematologic Diseases; Male; Prospective Studies; Reproducibility of Results; Reticulocyte Count; Retrospective Studies; Sensitivity and Specificity
PubMed: 21627508
DOI: 10.2460/javma.238.11.1452 -
Frontiers in Veterinary Science 2022Red blood cells (RBC) morphologic evaluation through microscopy optical (OM) and SEM, provides information to forecast, evaluate, and monitor the functioning of many...
Red blood cells (RBC) morphologic evaluation through microscopy optical (OM) and SEM, provides information to forecast, evaluate, and monitor the functioning of many organs. Factors, such aging and diseases affect RBC morphology in both, human and animals. SEM is useful to evaluate RBC morphology, although its use in diagnosis and evaluation in dogs is limited, due to the availability and cost. The aim of this research was to assess the normal RBC morphology in adult, senior and geriatrician dogs, clinically healthy by OM and SEM. In addition to evaluating the age effect, sex, body size, and their interaction on erythrocyte morphometry. To carry out the research 152 blood samples were evaluated from dogs of different sexes and body sizes (small, medium, and large). Three groups were made based on dogs age: group I adults (1-7.9 years old), group II senior (8-11.9 years old), and group III geriatricians (>12 years old). Erythrocyte parameters were evaluated by OM (diameter, height, and axial ratio). Per each dog, the parameters of 20 erythrocytes were measured. A total of 2,600 cells were scanned with the AmScope™ Software scale. In addition, the RBC morphology was evaluated by SEM. Statistical analyses used analysis of variance and a general linear model, which allows the comparison of multiple factors at two or more levels ( < 0.05). The results of this study showed that diameter and height were lower in adult dogs than in senior and geriatrician dogs ( < 0.05). Whereas, sex, body size, and the interaction did not show a significant effect ( > 0.05). Additionally, some images of anisocytosis, polychromasia, and poikilocytosis (echinocytes, acanthocytes, codocytes, spherocytes, stomatocytes, dacryocytes quatrefoil, and elliptocytes) were obtained by OM and SEM. Our study provides information about the morphological and morphometry alterations of adult, senior, and geriatrician dogs RBC. This work contributes to future investigations and the diagnosing diseases, where it is necessary to evaluate the morphology of RBC.
PubMed: 36439358
DOI: 10.3389/fvets.2022.998438 -
Cureus Mar 2022Sickle cell disease is an autosomal recessive disorder resulting in the substitution of CTG by CAG in the sixth codon of the beta-globin gene. As a result of this, the...
Sickle cell disease is an autosomal recessive disorder resulting in the substitution of CTG by CAG in the sixth codon of the beta-globin gene. As a result of this, the hydrophilic glutamic acid residue is replaced by hydrophobic valine residue, leading to the formation of hemoglobin tetramer HBS. This alteration in the beta-globin chain makes the red blood cells prone to sickling, especially in the presence of risk factors such as stress, hypoxia, and infection. These sickled red blood cells have the tendency to adhere to the endothelium and lead to vessel occlusion and distal tissue ischemia. The recent coronavirus disease 2019 (COVID-19) outbreak has impacted millions across the globe, putting individuals with co-morbidities at particularly high risk, and patients with sickle cell disease are no exception. We present the case of a 47-year-old African American male presenting to the emergency department with subjective fevers and a two-day history of pain in the arms, legs, and chest. A diagnosed case of sickle cell disease, the patient was on hydromorphone for pain management but ran out of his medications a few weeks before presentation. On examination, the patient was saturating well with mild tenderness upon palpation of the arms, legs, and chest. On complete blood count, the patient had a hemoglobin of 11.3 g/dL and a white cell count of 13.1 x10(3)/mcL. The patient had a normal mean corpuscular volume with reticulocytosis, hypochromia, ovalocytosis, poikilocytosis, polychromasia, and target cells. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) polymerase chain reaction (PCR) was positive. The chest X-ray did not reveal any significant findings. He was admitted to the medicine floor for the management of sickle cell crisis and was placed under airborne and droplet precautions. The patient was started on hydromorphone for pain management and intravenous fluid hydration. On the second day of admission, the patient reported increasing shortness of breath. He was saturating 90% on room air and 94% on 2 liters of supplemental oxygen. The white blood cell count increased to 18.42 x10(3)/mcL and the chest X-ray revealed reticular densities with patchy alveolar opacities in the left lung. Given the decline in respiratory status, the patient was started on remdesivir. Over the course of his hospital stay, the patient's pain and respiratory status improved, with the patient saturating 97% on room air. He was discharged home with instructions to follow isolation precautions for at least two weeks, folic acid, and adequate pain management. An appointment was also scheduled for the patient to follow with a sickle cell nurse practitioner upon discharge.
PubMed: 35494937
DOI: 10.7759/cureus.23604 -
Animals : An Open Access Journal From... Sep 2021Several extra-intestinal manifestations, including immune-mediated cytopenias, are reported in human inflammatory bowel disease (IBD), whereas they are poorly documented...
Several extra-intestinal manifestations, including immune-mediated cytopenias, are reported in human inflammatory bowel disease (IBD), whereas they are poorly documented in dogs. Hypothesizing that immune-mediated subclinical anemia can occur in canine IBD, the study aim was to evaluate the erythrogram and the presence of anti-RBC antibodies in dogs with IBD. IBD was diagnosed according to the following criteria: chronic gastrointestinal signs, ruling out of extra-intestinal diseases, no improvement with diet trial, histological evidence of inflammatory infiltration, and improvement after immunosuppressant therapy. Canine Chronic Enteropathy Clinical Activity Index (CCECAI) endoscopic and histopathological scores were assessed for each dog. Twenty-five dogs were enrolled, and each dog had a CBC evaluation prior to endoscopy. The CBC was performed using laser hematology analyzer and blood smears were carefully reviewed for the presence of nucleated RBC, anisocytosis, polychromasia, and Howell-Jolly bodies. IgG and IgM anti-RBC antibodies were evaluated with flow cytometry. A high frequency of positive cases for anti-RBC antibodies in dogs with IBD (17/25 dogs) was ascertained. Approximatively 50% of dogs showed some hematologic features of RBC regeneration in addition to hematologic findings consistent with chronic inflammation. Anti-RBC antibodies and signs of erythroid regeneration may suggest possible subclinical chronic immune-mediated hemolysis that can cause anemia in dogs with IBD, together with the chronic inflammation.
PubMed: 34573547
DOI: 10.3390/ani11092580 -
Comparative Medicine Dec 2016In the present study, we described the phenotype, histologic morphology, and molecular etiology of a mouse model of unstable hemoglobin Santa Ana. Hematologic evaluation...
In the present study, we described the phenotype, histologic morphology, and molecular etiology of a mouse model of unstable hemoglobin Santa Ana. Hematologic evaluation of anemic mice (Anem/+) discovered after N-ethyl-N-nitrosourea mutagenesis revealed moderate anemia with intense reticulocytosis and polychromasia, followed by anisocytosis, macrocytosis, hypochromia, and intraerythrocytic inclusion and Heinz bodies. The mice also demonstrated hemoglobinuria, bilirubinemia, and erythrocytic populations with differing resistance to osmotic lysis. Splenomegaly (particularly in older mutant mice) and jaundice were apparent at necropsy. Histopathologic examination revealed dramatically increased hematopoiesis and hemosiderosis in hematopoietic organs and intracellular iron deposition in tubular renal cells. These data are characteristic of a congenital hemolytic regenerative anemia, similar to human anemias due to unstable hemoglobin. Genetic mapping assigned the affected gene to mouse chromosome 7, approximately 50 cM from the Hbb locus. The sequence of the mutant Hbb gene exhibited a T→C transversion at nucleotide 179 in Hbb-b1, leading to the substitution of proline for leucine at amino acid residue 88 and thus homologous to the genetic defect underlying Santa Ana anemia in humans.
Topics: Anemia, Hemolytic, Congenital; Animals; Chromosome Mapping; Disease Models, Animal; Ethylnitrosourea; Female; Genotype; Hemoglobins, Abnormal; Humans; Male; Mice; Mice, Inbred BALB C; Mutation
PubMed: 28304246
DOI: No ID Found -
Retrospective and Prospective Investigations about "Quatrefoil" Erythrocytes in Canine Blood Smears.Veterinary Medicine International 2014The presence of unusual two RBCs patterns (so-called "quatrefoil RBCs," qRBCs) on canine blood smears at Optical Microscope (OM) was seen during routine evaluation of...
The presence of unusual two RBCs patterns (so-called "quatrefoil RBCs," qRBCs) on canine blood smears at Optical Microscope (OM) was seen during routine evaluation of CBCs. Two consecutive retrospective investigations were arranged including about 7,000 CBCs and clinical records and laboratory data from dogs showing qRBCs. Few samples with qRBCs were prepared for Scanning Electron Microscope (SEM). qRBCs were found in 6.89% (139 of 2016) and 8.47% (133 of 1569) of dogs and in 3.89% (154 of 3,958) and 4.47% (138 of 3,081) of CBCs (some dogs were tested more than once). Statistical analysis was significant for age groups (Chi squared, P < 0.0001), decreased total leukocyte and neutrophil counts (ANOVA, P < 0.0001), RBCs anisocytosis, polychromasia, and Howell-Jolly bodies (ANOVA, P < 0.018, <0.005, and <0.003, respectively). qRBCs were distributed in the area of feathered edge and at the smear side of body-feathered edge area in blood films. SEM ruled out the possibility of an optical illusion or an accidental overlap. qRBCs are associated with ageing of dogs, total leukocyte and neutrophil counts, and RBC anisocytosis, polychromasia, and Howell-Jolly bodies. Few hypotheses were discussed to explain the origin and meaning of this RBC arrangement.
PubMed: 24511413
DOI: 10.1155/2014/409573 -
PloS One 2022The Sysmex DI-60 digital morphology analyzer is a fully automated, cell-locating image analysis system. This study aimed to evaluate the analytical performance of DI-60.
BACKGROUND
The Sysmex DI-60 digital morphology analyzer is a fully automated, cell-locating image analysis system. This study aimed to evaluate the analytical performance of DI-60.
METHODS
A total of 822 peripheral blood smears were used. The diagnostic performance of DI-60 in terms of red blood cell (RBC) morphology characterization, white blood cell (WBC) differentials, and the total assay time including hands-on time was evaluated.
RESULTS
In comparison with manual slide review, DI-60 demonstrated acceptable accuracy in recognizing polychromasia, target cells, and ovalocytes. However, for schistocytes, DI-60 demonstrated low specificity (10.4%) despite the high sensitivity (97.2%). In the precision analysis of RBC morphology characterization, borderline samples harboring specific RBCs showed inconsistencies in the positive results among 20 replicates. Particularly, 6 of 10 samples showed inconsistencies in the precision for schistocytes. For WBC differentials, the overall agreement between pre-classification results and user-verified results was 89.4%. Except for basophils, normal WBCs showed a good correlation between DI-60 (after user verification) and manual counts. The sensitivities in detecting immature granulocytes, blasts, atypical lymphocytes, and normoblasts were 85.9%, 92.0%, 37.5%, and 77.6%, respectively. Although the total assay time of DI-60 was longer than that of manual review, the hands-on time was considerably shorter with a difference of 144.1 s/slide for abnormal samples.
CONCLUSION
DI-60 demonstrated acceptable performance for normal samples. However, for abnormal WBC differentials and RBC morphology characterization, it should be utilized carefully. DI-60 may contribute to an improvement in laboratory efficiency with increased feasibility.
Topics: Blood Cell Count; Erythrocyte Count; Hematologic Tests; Leukocyte Count; Leukocytes
PubMed: 35476704
DOI: 10.1371/journal.pone.0267638 -
Antimicrobial Agents and Chemotherapy Nov 2016Fungal infections pose a significant public health burden with high morbidity and mortality. CD101 is a novel echinocandin under development for the treatment and... (Comparative Study)
Comparative Study
Fungal infections pose a significant public health burden with high morbidity and mortality. CD101 is a novel echinocandin under development for the treatment and prevention of systemic Candida infections. Preclinical studies were conducted to evaluate the metabolic stability, plasma protein binding, pharmacokinetics, toxicity, and efficacy of CD101 at various dose levels. CD101 was stable to biotransformation in rat, monkey, and human liver microsomes and rat, monkey, dog, and human hepatocytes. In vitro studies suggest minimal interaction with recombinant cytochrome P450 enzymes (50% inhibitory concentrations [ICs] of >10 μM). Similar to anidulafungin, CD101 bound avidly (>98%) to human, mouse, rat, and primate plasma proteins. In a 2-week repeat-dose comparison study, CD101 was well tolerated in rats (no effects on body weight, hematology, coagulation, or urinalysis). In contrast, administration of anidulafungin (at comparable exposure levels) resulted in reduced body weight, decreases in red blood cell, hemoglobin, hematocrit, mean cell volume, mean corpuscular hemoglobin, platelet, and reticulocyte counts, increases in neutrophil and eosinophil counts, polychromasia, and decreased activated partial thromboplastin time. Elevated plasma transaminases, total bilirubin, cholesterol, and globulin, dark and enlarged spleens, and single-cell hepatocyte necrosis were also observed for anidulafungin but not CD101. Hepatotoxicity may be due to the inherent chemical lability of anidulafungin generating potentially reactive intermediates. A glutathione trapping experiment confirmed the formation of a reactive species from anidulafungin, whereas CD101 did not exhibit instability or reactive intermediates. CD101 showed antifungal activity against Candida and Aspergillus infections in neutropenic mice. These preclinical studies demonstrated that CD101 is chemically and metabolically stable, well tolerated with no hepatotoxicity, and efficacious as an antifungal agent.
Topics: Anidulafungin; Animals; Antifungal Agents; Aspergillosis; Biotransformation; Blood Proteins; Candidiasis; Cytochrome P-450 Enzyme Inhibitors; Dose-Response Relationship, Drug; Echinocandins; Female; Humans; Inactivation, Metabolic; Macaca fascicularis; Male; Microbial Sensitivity Tests; Microsomes, Liver; Rats, Sprague-Dawley
PubMed: 27620474
DOI: 10.1128/AAC.00701-16