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Biotechnic & Histochemistry : Official... Feb 2011An introduction to the nomenclature and concept of "Romanowsky stains" is followed by a brief account of the dyes involved and especially the crucial role of azure B and... (Review)
Review
An introduction to the nomenclature and concept of "Romanowsky stains" is followed by a brief account of the dyes involved and especially the crucial role of azure B and of the impurity of most commercial dye lots. Technical features of standardized and traditional Romanowsky stains are outlined, e.g., number and ratio of the acidic and basic dyes used, solvent effects, staining times, and fixation effects. The peculiar advantages of Romanowsky staining are noted, namely, the polychromasia achieved in a technically simple manner with the potential for stain intensification of "the color purple." Accounts are provided of a variety of physicochemically relevant topics, namely, acidic and basic dyeing, peculiarities of acidic and basic dye mixtures, consequences of differential staining rates of different cell and tissue components and of different dyes, the chemical significance of "the color purple," the substrate selectivity for purple color formation and its intensification in situ due to a template effect, effects of resin embedding and prior fixation. Based on these physicochemical phenomena, mechanisms for the various Romanowsky staining applications are outlined including for blood, marrow and cytological smears; G-bands of chromosomes; microorganisms and other single-cell entities; and paraffin and resin tissue sections. The common factors involved in these specific mechanisms are pulled together to generate a "universal" generic mechanism for these stains. Certain generic problems of Romanowsky stains are discussed including the instability of solutions of acidic dye-basic dye mixtures, the inherent heterogeneity of polychrome methylene blue, and the resulting problems of standardization. Finally, a rational trouble-shooting scheme is appended.
Topics: Azure Stains; Carmine; Chemical Phenomena; Coloring Agents; Eosine I Bluish; Eosine Yellowish-(YS); Fluoresceins; Histological Techniques; Hydrogen-Ion Concentration; Phenothiazines; Solvents
PubMed: 21235292
DOI: 10.3109/10520295.2010.515491 -
Pathology 2009To study the cytological features of breast lesions associated with hydrophilic polyacrylamide gel (PAAG) injection augmentation mammoplasty and to determine the...
AIMS
To study the cytological features of breast lesions associated with hydrophilic polyacrylamide gel (PAAG) injection augmentation mammoplasty and to determine the specific diagnostic features.
METHODS
Fine needle aspiration (FNA) cytology smears from 14 patients who presented with breast lump and a clinical history of PAAG injection were reviewed. The staining properties of the PAAG material in Papanicolaou (Pap), Haematoxylin and Eosin (H&E) and Diff-Quik stains, as well as the cellular background, were studied. Cell blocks were also studied with H&E, periodic acid-Schiff (PAS), periodic acid-Schiff with diastase digestion (PASD) and mucicarmine (MC) stains.
RESULTS
PAAG was stained consistently pinkish with a homogenous smooth texture on H&E (100%), polychromasia on Pap (100%) and magenta violet with bubbly vacuoles on Diff-Quik (100%). Half (7) of the cases showed sharp borders. The presence of foreign body type giant cells with histiocytes (5 cases, 36%) and scanty clusters of benign ductal cells (4 cases, 29%) were seen in the background of a minority of the cases. PAAG was represented by blocks of homogenous grey-purplish acellular material with a sharp border on cell block sections, which were negative for PAS, PASD and MC.
CONCLUSION
PAAG shows a consistent staining pattern in various common cytological staining preparations. Blocks of acellular gelatinous material, stained homogenous pinkish on H&E, polychromasia on Pap and magenta violet with bubbly vacuoles on Diff-Quik, are the reliable cytological features of PAAG.
Topics: Acrylic Resins; Adult; Biopsy, Fine-Needle; Breast Diseases; Female; Humans; Mammaplasty
PubMed: 19900083
DOI: 10.1080/00313020903041168 -
Journal of Veterinary Emergency and... Oct 2009The purpose of this study was to determine the LD(50) for acute blood loss in mallard ducks (Anas platyrhynchos), compare the mortality rate among 3 fluid resuscitation... (Randomized Controlled Trial)
Randomized Controlled Trial
OBJECTIVE
The purpose of this study was to determine the LD(50) for acute blood loss in mallard ducks (Anas platyrhynchos), compare the mortality rate among 3 fluid resuscitation groups, and determine the time required for a regenerative RBC response.
DESIGN
Prospective study.
SETTING
Medical College of Wisconsin Research facility.
ANIMALS
Eighteen mallard ducks were included for the LD(50) study and 28 for the fluid resuscitation study.
INTERVENTIONS
Phlebotomy was performed during both the LD(50) and fluid resuscitation studies. Ducks in the fluid resuscitation study received a 5 mL/kg intravenous bolus of crystalloids, hetastarch (HES), or a hemoglobin-based oxygen-carrying solution (HBOCS).
MEASUREMENTS AND MAIN RESULTS
The LD(50) for acute blood loss was 60% of total blood volume. This blood volume was removed in the fluid resuscitation study to create a model of acute blood loss. Following fluid administration, 6 birds in the crystalloid group (66%), 4 birds in the HES group (40%), and 2 birds in the HBOCS group (20%) died. No statistical difference in mortality rate was seen among the 3 fluid resuscitation groups. Relative polychromasia evaluated post-phlebotomy demonstrated regeneration starting at 24 hours and continuing through 48 hours.
CONCLUSIONS
The LD(50) for acute blood loss in mallard ducks was 60% of their total blood volume. Although no statistical difference in mortality rate was appreciated among the 3 fluid resuscitation groups, a trend of decreased mortality rate was observed in the HBOCS group. An early regenerative response was apparent following acute blood loss.
Topics: Animals; Crystalloid Solutions; Ducks; Female; Fluid Therapy; Hemoglobins; Hemorrhage; Hydroxyethyl Starch Derivatives; Isotonic Solutions; Male; Shock, Hemorrhagic
PubMed: 19821888
DOI: 10.1111/j.1476-4431.2009.00465.x -
Veterinary Clinical Pathology Jun 2009Hematologic analysis is an important tool in evaluating the general health status of free-ranging manatees and in the diagnosis and monitoring of rehabilitating animals.
BACKGROUND
Hematologic analysis is an important tool in evaluating the general health status of free-ranging manatees and in the diagnosis and monitoring of rehabilitating animals.
OBJECTIVES
The purpose of this study was to evaluate diagnostically important hematologic analytes in healthy manatees (Trichechus manatus) and to assess variations with respect to location (free ranging vs captive), age class (small calves, large calves, subadults, and adults), and gender.
METHODS
Blood was collected from 55 free-ranging and 63 captive healthy manatees. Most analytes were measured using a CELL-DYN 3500R; automated reticulocytes were measured with an ADVIA 120. Standard manual methods were used for differential leukocyte counts, reticulocyte and Heinz body counts, and plasma protein and fibrinogen concentrations.
RESULTS
Rouleaux, slight polychromasia, stomatocytosis, and low numbers of schistocytes and nucleated RBCs (NRBCs) were seen often in stained blood films. Manual reticulocyte counts were higher than automated reticulocyte counts. Heinz bodies were present in erythrocytes of most manatees. Compared with free-ranging manatees, captive animals had slightly lower MCV, MCH, and eosinophil counts and slightly higher heterophil and NRBC counts, and fibrinogen concentration. Total leukocyte, heterophil, and monocyte counts tended to be lower in adults than in younger animals. Small calves tended to have higher reticulocyte counts and NRBC counts than older animals.
CONCLUSIONS
Hematologic findings were generally similar between captive and free-ranging manatees. Higher manual reticulocyte counts suggest the ADVIA detects only reticulocytes containing large amounts of RNA. Higher reticulocyte and NRBC counts in young calves probably reflect an increased rate of erythropoiesis compared with older animals.
Topics: Animals; Blood Cell Count; Eosinophils; Erythrocytes; Hematologic Tests; Trichechus manatus
PubMed: 19490571
DOI: 10.1111/j.1939-165X.2009.00113.x -
Blood Cells, Molecules & Diseases 2009One of the most commonly inherited anemias in man is Hereditary Spherocytosis (HS) with an incidence of 1 in 2000 for persons of Northern European descent. Mouse models...
One of the most commonly inherited anemias in man is Hereditary Spherocytosis (HS) with an incidence of 1 in 2000 for persons of Northern European descent. Mouse models of HS include spontaneous inherited hemolytic anemias and those generated by gene targeting. The Neonatal anemia (Nan) mouse is a novel model of HS generated by N-ethyl-N-nitrosurea mutagenesis and suffers from a severe neonatal anemia. Adult Nan mice have a lifelong hemolytic anemia with decreased red blood cell numbers, hematocrit, and hemoglobin, but elevated zinc protoporphyrin levels. Blood smears taken from Nan mice show a hypochromic anemia characterized by poikilocytosis, anisocytosis and polychromasia. The Nan phenotype can be transferred by bone marrow transplantation indicating that the defect is intrinsic to bone marrow. The hemolytic anemia in adult Nan mice can be identified by osmotic fragility testing. Examination of the erythrocyte membrane skeleton proteins (EMS) reveals a global deficiency of these proteins with protein 4.1a being completely absent. The Nan locus maps to mouse Chromosome 8 and does not co-localize with any known EMS genes. The identification of the Nan gene will likely uncover a novel protein that contributes to the stability of the EMS and may identify a new mutation for HS.
Topics: Anemia, Hemolytic; Animals; Bone Marrow Transplantation; Chromosome Mapping; Chromosomes, Mammalian; Disease Models, Animal; Erythrocyte Membrane; Erythrocytes; Male; Membrane Proteins; Mice; Mice, Inbred C57BL; Microscopy, Electron, Scanning
PubMed: 19409822
DOI: 10.1016/j.bcmd.2009.03.009 -
Journal of Virology Apr 2009The genome organization of the novel human papillomavirus type 108 (HPV108), isolated from a low-grade cervical lesion, deviates from those of other HPVs in lacking an...
The genome organization of the novel human papillomavirus type 108 (HPV108), isolated from a low-grade cervical lesion, deviates from those of other HPVs in lacking an E6 gene. The three related HPV types HPV103, HPV108, and HPV101 were isolated from cervicovaginal cells taken from normal genital mucosa (HPV103) and low-grade (HPV108) and high-grade cervical (HPV101) intraepithelial neoplasia (Z. Chen, M. Schiffman, R. Herrero, R. DeSalle, and R. D. Burk, Virology 360:447-453, 2007, and this report). Their unusual genome organization, against the background of considerable phylogenetic distance from the other HPV types usually associated with lesions of the genital tract, prompted us to investigate whether HPV108 E7 per se is sufficient to induce the above-mentioned clinical lesions. Expression of HPV108 E7 in organotypic keratinocyte cultures increases proliferation and apoptosis, focal nuclear polymorphism, and polychromasia. This is associated with irregular intra- and extracellular lipid accumulation and loss of the epithelial barrier. These alterations are linked to HPV108 E7 binding to pRb and inducing its decrease, an increase in PCNA expression, and BrdU incorporation, as well as increased p53 and p21(CIP1) protein levels. A delay in keratin K10 expression, increased expression of keratins K14 and K16, and loss of the corneal proteins involucrin and loricrin have also been noted. These modifications are suggestive of infection by a high-risk papillomavirus.
Topics: Adult; Apoptosis; Cell Proliferation; Cell Transformation, Neoplastic; DNA, Viral; Female; Genome, Viral; Humans; Keratinocytes; Lipid Metabolism; Papillomaviridae; Papillomavirus E7 Proteins; Papillomavirus Infections; United States; Uterine Cervical Neoplasms
PubMed: 19153227
DOI: 10.1128/JVI.02490-08 -
Veterinary Clinical Pathology Jun 2008A 1-year-old intact male Boxer was presented to the Texas Veterinary Medical Center for emergency treatment following suspected ingestion of a large number of tablets of...
A 1-year-old intact male Boxer was presented to the Texas Veterinary Medical Center for emergency treatment following suspected ingestion of a large number of tablets of Adderall, a pharmaceutical amphetamine. The dog had a temperature of 41.7 degrees C, heart rate of 192 beats per minute, and a respiratory rate of 100 breaths per minute. The dog was anxious and agitated with bilaterally dilated pupils, and shortly thereafter became recumbent and incontinent. Initial CBC results included mild leukopenia and mild thrombocytopenia. The dog was not anemic (HCT 39.9%) and had only slight polychromasia, but had 48 nucleated RBCs/100 WBC (7500/microL). Moderate numbers of neutrophils had hypersegmented nuclei and several pyknotic cells were noted. The metarubricytosis persisted for approximately 56 hours while hypersegmentation and pyknotic cells were no longer found at 8 hours after presentation. The dog received supportive care and recovered uneventfully. We hypothesized that hyperpyrexia associated with Adderall toxicity resulted in inappropriate metarubricytosis due to damaged bone marrow endothelium, and resulted in hypersegmentation and pyknosis due to damaged or accelerated aging of neutrophils in peripheral blood. Metarubricytosis has been reported previously in dogs with heat-induced illness, such as heat stroke.
Topics: Acid-Base Equilibrium; Alkaline Phosphatase; Amphetamines; Animals; Blood Proteins; Central Nervous System Stimulants; Chlorides; Creatine Kinase; Dog Diseases; Dogs; Hypercholesterolemia; Magnesium; Male; Potassium; Sodium; Treatment Outcome
PubMed: 18533918
DOI: 10.1111/j.1939-165X.2008.00031.x -
Medicina Clinica Apr 2008Unstable hemoglobins (Hb) show amino acid substitutions in critical places that produce a decrease of molecular solubility facilitating its denaturalization and...
BACKGROUND AND OBJECTIVE
Unstable hemoglobins (Hb) show amino acid substitutions in critical places that produce a decrease of molecular solubility facilitating its denaturalization and precipitation. We describe the first case of Hb Newcastle in Spain.
CASE REPORT
5 year-old girl who came to visit due to fever over 4 days. Physical examination disclosed pale skin with subicteral mucosaes and splenomegaly. Lab analysis disclosed: Hb, 79 g/l; haematocrit, 0.27 l/l, mean corpuscular volume 93.4 fl, and reticulocyte count of 3%, along with anysocytosis and polychromasia.
RESULTS
Hemoglobin heat stability test was positive. Hemoglobin electrophoresis showed a low band at HbA2. High performance liquid chromatography (HPLC) showed an Hb peak corresponding to the 12% of total Hb. Beta globin gene sequentiation showed the CD92 His --> Pro mutation Hb Newcastle in heterocygote condition in patient and her mother.
CONCLUSIONS
Hb Newcastle has been described in 3 patients of English, Russian and Chinese origin. Clinical manifestation is chronic hemolytic anemia with severe crisis after oxidant drugs ingestion or infections. By using the electrophoretic method, a diffuse pattern of Hb bands between HbA and HbA2 is observed, difficulting the precise identification of the abnormal Hb. This inconvenience is overcomed by using HPLC that allows the clear identification of the abnormal Hb Newcastle.
Topics: Catchment Area, Health; Child, Preschool; Chromatography, High Pressure Liquid; Diagnosis, Differential; Ethnicity; Female; Hemoglobinopathies; Humans; Spain
PubMed: 18405501
DOI: 10.1157/13118108 -
American Journal of Therapeutics 2008Oxcarbazepine is a commonly used antiepileptic and mood stabilizer medication with a considerably good safety profile. Medication-induced side effects are a common cause...
Oxcarbazepine is a commonly used antiepileptic and mood stabilizer medication with a considerably good safety profile. Medication-induced side effects are a common cause of morbidity in the geriatric population. We describe a patient presenting with hemolytic anemia attributed to oxcarbazepine that resolved after discontinuation of the drug. A 75-year-old male was brought into the hospital because of lethargy for the previous few days. His medical history included diet-controlled diabetes mellitus, mild essential hypertension, ischemic stroke with no residual deficit, seizure disorder, and dementia. He was taking only trileptal for the previous 3 months. His other history was insignificant. Physical examination was remarkable for generalized pallor and impaired memory. Laboratory results showed hemoglobin of 4.6 g/dL. Serum lactate dehydrogenase was 1314 u/L, and total bilirubin was 3.4 mg/dL, with indirect bilirubin of 2.9 mg/dL. Peripheral blood smear revealed microspherocytes, polychromasia, and a few nucleated red blood cells. Urinalysis showed 12RBCs/high power field (HPF), hemosiderinuria, and moderate bilirubin. Coombs test, cold agglutinins, antinuclear antibodies, and cultures were negative. Computerized tomographic scan of the chest, abdomen, and pelvis was also normal. After exclusion of other causes of hemolysis, drug-induced hemolytic anemia was suspected, and oxcarbazepine was discontinued. The patient's symptoms and laboratory parameters improved. He was in a usual state of health at 7 months follow-up. To our knowledge, this is the first reported case of oxcarbazepine-induced hemolytic anemia in the medical literature, and it was reported to the manufacturing pharmaceutical company.
Topics: Aged; Anemia, Hemolytic; Anticonvulsants; Carbamazepine; Epilepsy; Humans; Male; Oxcarbazepine
PubMed: 18356642
DOI: 10.1097/MJT.0b013e31815afb6f -
Blood Reviews Jan 2008The diagnosis of autoimmune hemolytic anemia (AHA) requires evidence of shortened red blood cell (RBC) survival mediated by autoantibodies directed against autologous... (Review)
Review
The diagnosis of autoimmune hemolytic anemia (AHA) requires evidence of shortened red blood cell (RBC) survival mediated by autoantibodies directed against autologous RBCs. About 80 percent of patients with AHA have warm-reactive antibodies of the IgG isotype; the remainder exhibit cold-reactive autoantibodies. Typical patients exhibit anemia, reticulocytosis, spherocytes and polychromasia on the blood film and a positive direct antiglobulin test (DAT). Increased indirect serum bilirubin, urinary urobilinogen and serum lactate dehydrogenase (LDH), and decreased serum haptoglobin are not required for the diagnosis, but are frequently present. Patients with AHA and no underlying associated disease are said to have primary or idiopathic AHA. AHA in patients with associated autoimmune disease and certain malignant or infectious diseases is classified as secondary. The etiology of AHA is unknown. Patients with symptomatic anemia require transfusion of RBCs. Prednisone and splenectomy may provide long term remission. Rituximab, intravenous immunoglobulin, immunosuppressive drugs and danazol have been effective in refractory cases and for patients who are poor candidates for surgery.
Topics: Anemia, Hemolytic; Autoantibodies; Blood Transfusion; Coombs Test; Humans; Immunoglobulin G; Immunosuppressive Agents; Spherocytes; Splenectomy; Temperature
PubMed: 17904259
DOI: 10.1016/j.blre.2007.08.001