Benign Heme Flashcards

Question

Heinz bodies

Answer 1

Ppt hemoglobin, requires supravital dye to visualize (seen in G6PD deficiency and with unstable forms of Hgb)

Answer 2

Remnant of nuclear DNA (seen in post splenectomy, in hemolytic anemia, or in megaloblastic anemia)

Answer 3

Seen in iron deficiency anemia, thalassemia, and sideroblastic anemia

Answer 4

Seen with increased erythropoiesis (due to increased reticulocytes), megaloblastic anemia (oval macrocytes), and liver disease (round macrocytes)

Answer 5

Seen in iron deficiency anemia, thalessemia, and sideroblastic anemia

Answer 6

Iron containing mitochondria (seen in sideroblastic anemia and post splenectomy)

Answer 7

Hypochromic variants of elliptocytes that have long axis at least three times the length of the short axis (classic for Fe deficiency but also seen in anemia of chronic disease and beta thalassemia minor

Answer 8

Due to residual ribosomal material

Answer 9

Seen in sickling hemoglobinopathies (NOT seen in sickle cell trait)

Answer 10

Seen in hereditary spherocytosis and immunohemolytic anemia

Answer 11

Usually artifact; may be seen in alcoholism, hereditary stomatocytosis, or Rh null phenotype

Answer 12

Liver disease, HbC, SC disease, HbE, post splenectomy, thalassemia

Answer 13

Myelophthisic dz, splenic dysfunction

Answer 14

Thought to be remnants of rough ER, possibly agglutinated ribosomes

Answer 15

Prominent primary granules; seen in sepsis and GM-CSF (Neupogen) therapy

Answer 16

Larger than lymphocytes (2x) with a moderate amount of cytoplasm, round nuclei (occasionally irregular), immature chromatin, and prominent central nucleolus

Answer 17

May be T or NK phenotype; very common to be increased post transplant

Answer 18

Use supravital stain (eg new methylene blue)

Answer 19

RNA binding dyes that can be detected by fluorescence (but Coulter still uses detection of light scatter after staining with new methylene blue)

Answer 20

Cells with the highest fluorescence intensity (highest RNA content)

Answer 21

% retics (# per 100 RBCs)

Answer 22

(retic%)(hct/45) -takes into account normal RBC production for a given hematocrit

Answer 23

CRC/correction factor -1.0 for hct 40-45 -1.5 for hct 35-39 -2.0 for hct 25-34 -2.5 for hct 15-24 -3.0 for hct

Answer 24

Usually present in blood for 24 hours before they extrude residual RNA and become erythrocytes; if released early from marrow, immature retics may persist in PB for 2-3 days (RPI corrects for presence of immature retics)

Answer 25

failure of BM RBC production (hypo proliferative anemia)

Answer 26

marrow hyperproliferation or appropriate response to anemia

Answer 27

Fanconi anemia Dyskeratosis congenita Diamond-Blackfan anemia Congenital amegakaryocytic thrombocytopenia Schwachman-Diamond syndrome Severe congenital neutropenia (Kostman syndrome)

Answer 28

-PNH -Idiopathic: (theory: destruction of CD34+ progenitor cells through apoptotic mechanisms stimulated by cytokine released from activated BM cytotoxic T cells) -Secondary to drugs (cytotoxic drugs, chloramphenicol, gold), radiation, toxins (benzen), infections (parvovirus), neoplasms (thymoma), pregnancy

Answer 29

Diamond-Blackfan anemia

Answer 30

P-parvovirus infection I- idiopathic T- transient erythroblastopenia of childhood (most likely due to antecedent viral infection) A- antierythropoietin ab induced red cell aplasia (occurs in some patients receiving recombinant epo) S- sustained pure red cell aplasi (secondary to neoplasms such as T cell LGL leukemia, immune d/o, chronic viral infections, and drugs)

Answer 31

-AR (rarely X-liked recessive) -13 genes on many different chromosomes (one being BRCA2) -cells characteristically show abnormally high frequency of spontaneous chromosomal breakage and hypersensitivity to DNA cross-linking agents such as diepoxybutane (DEB) and mitomycin C

Answer 32

increased chromosomal breakage seen in FA cells compared to normal controls after exposure to DEB or mitomycin C (AKA the DEB/MMC stress test)

Answer 33

-progressive BM failure (aplastic anemia with pancytopenia) and increased risk of malignancy (AML- often with monocytic features, solid tumors like cutaneous malignancies, HCC, gastric carcinoma) -cafe au lait spots, microcephaly, hypoplastic thumbs, absent radius, scoliosis, MR, horseshoe kidney, duodenal atresia, cardiac defects, neurologic abnormalities (1/3 no abnormalities) -present toward end of 1st decade -anabolic steroids (oxymetholone) produce useful trilineage hematopoietic response in 50-70%, but most become refractory -tx HSCT

Answer 34

-X-linked recessive (80% cases), but AD and AR subtypes are recognized -DKC1 gene on Xq28 -genomic instability d/o similar to Fanconi anemia (however, no increased chromosomal breakage)

Answer 35

Inherited BM failure syndrome characterized by mucocutaneous triad (mostly males): 1. abnormal skin pigmentation 2. nail dystrophy 3. oral leukoplakia -skin and nail changes appear first, usually by age 10 -BM failure by age 20 -like Fanconi anemia, main cause of death is BM failure -other causes of death include pulmonary complications and malignancy -again, like FA, anabolic steroids (oxymetholone) produce useful trilineage hematopoietic response in 50-70% -tx HSCT

Answer 36

-AR, 90% due to mutation of SBDS gene (chr 7q11) -results in deficiency of SBDS protein (can see loss by IHC) -protein plays regulatory role in RNA metabolism in nucleolus by stabilizing mitotic spindle -can see an isochromosome 7q

Answer 37

-2nd most common cause of pancreatic insufficiency in childhood after CF -exocrine pancreatic insufficiency (86%), BM failure-mostly as neutropenia (98%), anemia (42%), pancytopenia (19%), skeletal abnormalities (50% particularly metaphyseal dysostosis), growth retardation (56%), and increased risk of MDS/AML (30%) -tx with pancreatic enzymes and G-CSF -main cause of death infection/bleeding -at risk for complications from HSCT

Answer 38

-DBA1 gene AKA RPS19 - encodes ribosomal protein S19 and is NOT a regulator of erythropoiesis rather is a ubiquitous ribosomal protein -AD with incomplete penetrance -heterozygosity for mutations of gene (genetic heterogeneity of DBA)

Answer 39

-presents in early infancy (around 8 weeks) with sx of anemia -hallmark is selective decrease in erythroid precursors and normochromic macrocytic anemia -craniofacial, thumb, cardiac, and urogenital malformations -modest increase in MDS/AML/osteosarcoma -tx with corticosteroids and transfusions; HSCT

Answer 40

DB: pure RBC aplasia SD: neutropenia FA: pancytopenia

Answer 41

-elevated erythrocyte deaminase activity and elevated fetal hemoglobin (HbF) -i ag overexpressed on RBCs

Answer 42

1. normochromic, macrocytic anemia developing in early childhood 2. reticulocytopenia 3. normocellular BM with marked selective deficiency of eyrthroid precursors (erythroblasts

Answer 43

clumped ribosomal RNA (not dots-those are Heinz bodies)

Answer 44

- On right, RBC contains 4 small, red-purple dots (tetrad) - On left, 2 RBCs contain trophozoites (ring forms), which closely resemble P. falciparum - RBCs may show single chromatic dot or two or more chromatic masses; occasionally 4 ring forms in tetrad =\> maltese cross - 4 visible dots is diagnostic and not seen in malaria; can also see extraeythrocytic forms which are not seen in malaria - in malaria, RBCs are enlarged/ovoid vs babesia they are normal - noncyclic fever and hemolysis - coinfection with Lyme dz and flavivirus

Answer 45

- allergic/hypersensitivity reactions - inflammatory d/o (collagen vascular dz, RA, UC) - endocrinopathy (DM) - chronic renal failure - infections (influenza, chicken pox) - neoplasms (CML)

Answer 46

AR - PMNs impaired mobility (defective chemotaxis) and defective granulation - primary defect in intracellular granules: \*leads to WBC death in marrow resulting in neutropenia and thrombocytopenia \*defective granules in grans, lymphs, and NK cells alter immune fx with resultant severe pyogenic infections \*abnormal melanin containing granules result in eye/skin hypopigmentation - all WBCs contain large cytoplasmic granules (due to fusion of lysosomes) - cytoplasmic inclusions contain both primary (MPO+) and secondary granules

Answer 47

- severe recurrent bacteria infections - partial oculocutaneous albinism - many develop EBV associated hemophagocytic syndrome ("accelerated phase") or EBV induced lymphoproliferative d/o - pts die in childhood - tx HSCT

Answer 48

-rare disorders characterized by profound morphologic abnormalities of erythroid elements and ineffective erythropoiesis - some cases are AR - all pts have anemia with marked anisopoikilocytosis, reticulocytopenia, marrow erythroid hyperplasia

Answer 49

- erythroid precursors show megaloblastic changes with internuclear chromatin bridges - occasional binucleate forms seen

Answer 50

\*\*The most common type\*\* - characterized by bi- and multinucleated erythroid precursors (usually 2-7 nuclei) - positive acidified serum test (Ham's test) due to an abnormal RBC antigen; lysis occurs in heterologoys serum only, in contrast to PNH in which lysis occurs in both autologous and heterologous serum - also referred to as "hereditary erythroblastic multinuclearity with positive acidified serum" HEMPAS - RBCs have very high density of i (recall Diamond-Blackfan too)

Answer 51

- allerigic/atopic disorders - medications (L-tryptophan linked to eosinophilia-myalgia syndrome) - parasites (must invade tissue to produce eosinophilia) - cutaneous disorders (eczema, atopic dermatits, bullous pemphigoid) - inflammatory disorders (IBD, collagen vascular dz, sarcoid, celiac) - pulmonary disorders (Loeffler syndrome, CF, Churg-Strauss) - neoplasms (T cell lymphoproliferative d/o, HL, LCH)

Answer 52

-pronounced multinucleateion with up to 12 nuclei present in some erythroid precursors ("gigantoblasts")

Answer 53

Release of secondary granules: \*major basic protein \*peroxidase \*arylsulfatase \*histamine oxidase \*eosinophil cationic protein - modulate immediate hypersensitivity rxn - destroy parasites

Answer 54

- Two isoforms of spectrin,  and , form a loosely wound helix. Two  helices are linked end to end to form a tetramer, which has binding sites for several other proteins, including other spectrin molecules. - The spectrin tetramers are organized into a meshwork that is fixed to the membrane by the protein ankyrin. - Ankyrin is itself connected to a transmembrane protein called Band 3 (AE1, anion exchanger). - Band 4.2 (palladin) is thought to function to stabilize the link between ankyrin and Band 3. - Spectrin is also linked to the transmembrane protein glycophorin C by the protein known as Band 4.1. - Thus the meshwork is anchored to the membrane at multiple sites. - Band 4.1 stabilizes the association of spectrin with actin, as does the protein adducin. - Actin subunits form short microfilaments consisting of actin and tropomyosin. - The protein tropomodulin is also associated with filamentous actin.

Answer 55

- useful in monitoring warfarin in presence of a LA or DTI - utilizes an activator of factor X and peptide substrate for factor Xa that is coupled to a chromogenic marker - cleavage of peptide by factor Xa releases the marker and yields a colored product, the intensity of which is directly proportional to factor X levels

Answer 56

- serum level has a direct correlation with total amount of iron stored in the body (less invasive than BM bx) - is a storage complex of iron and apoferritin protein - ferritin is an acute phase reactant and often increase with infection, inflammation, or malignancy - normal is 10-500 ng/mL

Answer 57

- measures amount of circulating iron that is bound to transferrin - serum iron concentrates show wide diurnal variation; should collect specimen in AM when level is highest - normal is 60-180 µg/dL

Answer 58

- measures the extent to which iron-binding sites in serum can be saturated - becuase iron-binding sites in serum are almost entirely on circulating transferrin, this is really an indirect measurement of the amount of transferrin in the blood - normal is 250-410 µg/dL

Answer 59

(serum Fe/TIBC) x 100 normal is approximately 30% (20-50%)

Answer 60

UIBC= TIBC-serum iron

Answer 61

- transferrin receptor is a transmembrane protein that transfers iron from plasma transferrin into the RBC - most transferrin receptors are found in the cell membrane, but a portion is found in soluble form in the serum - transferrin receptor levels reflect iron status, with receptor synthesis being rapidly induced by decreased iron levels; unlike ferritin it is not altered in inflammatory states - useful to distinguish iron deficiency (increased) from thalassemia minor (normal) and anemia of chronic disease (normal to slightly increased) - elevated levels may also be seen in pts with hematolymphoid neoplasms or conditions with increased hematopoiesis (eg HA, hemoglobinopathies, folate/B12 def) irrespective of iron status

Answer 62

(zinc protoporphyrin is similar) - FEP levels are increased in pts with anemias associated with failure of iron incorporation into heme - when insufficient iron is available for developing erythroblasts, excess protoporphyrin that was destined to be converted to heme accumulates as FEP - FEP is high in iron deficiency anemia and in anemias associated with block in iron utilization (anemia of chronic dz, lead poisoning, sideroblastic anemia); FEP is normal in thalassemia (in thalassemia, globin synthesis is abnormal but heme synthesis is normal) - useful to distinguish iron deficiency (increased) from thalessemia minor - normal less than 100 µg/dL

Answer 63

- iron is stored in reticuloendothelial cells - erythroid precursors that cotnain one or more particles of stainable iron are known as sideroblasts - iron is stored as ferritin (iron complexed to the apoferritin protein) and hemosiderin (iron-protein complexes with a high iron content and denatured ferritin aggregates); hemosiderin is visible with Prussian blue stain - when storage iron is present in the BM, anemia cannot be a result of iron deficiency (unless the pt has recently received parenteral iron) - normally 20-40% of erythroid precursors are sideroblasts

Answer 64

- G6PD reduces NADP+ to NADPH while oxidizing glucose-6-phosphate in the hexose monophosphate shunt - NADPH is then used to convert oxidized glutathione to reduced glutathione - reduced glutathione protects RBCs from oxidizing injury by catalyzing breakdown of oxidants such as H2O2 (O free radicals) - deficiency of G6PD leads to insufficient glutathione, which renders RBCs susceptible to oxidant injury - oxidative agents include free radicals generated during an infection or following exposure to various drugs (sulfonamides, nitrofurantoin, antimalarials) - \>400 mutations of G6PD gene are known; the two most common are the A variant, which occurs in 12% of AA males and the Mediterranean variant, which is more severe and can occur after ingesting fava beans

Answer 65

- X linked recessive (the enzyme deficiency is present in all RBCs of affected males) - heterozygous females have 2 populatins of RBCs (normal and G6pD deficient); the proportion of each is dependant on random inactivation of the X chromosomes (some patients healthy vs others appear fully affected)

Answer 66

- hemolysis begins acutely in infection and 1-3 days after drug/fava bean ingestion - hemolysis often severe, leading to hemoglobinemia, hemoglobinuria, and jaundice - G6PD-A deficiency the hemolytic anemia is usually self limited b/c reticulocytes have nearly normal G6PD levels; more severe forms may require transfusions

Answer 67

PB smear: Heinz bodies on supravital stain (ppt hemoglobin) or bite cells Fluorescent spot test (screening test): * NADPH fluoresces when exposed to UV light; NADP+ does not * RBCs mixed with soln of G6P, oxidized glutathione, NADP+ which is then spotted on filter paper * Normally, when the spot is exposed to UV light, fluorescence appears within 5-10 min * G6PD deficient samples, no fluorescence seen - screening test may not be useful in females or post-hemolytic period (reticus more G6PD activity) - confirmation of abnormal screening requires quantitative assay of G6PD based on spectrophotometric quantification of NADPH formation from NADP+

Answer 68

- most common lysosomal storage dz - Ashkenazi Jewish pop (1 in 450; 1 in 100K of general pop) - AR inheritance, mutation in glucocerebrosidase gene (GBA) with \>200 mutations described - defective function of ß-glucocerebrosidase

Answer 69

PB: moderate-marked anisopoikilocytosis, mild microcytosis, hypochromasia, moderate thrombocytopenia BM: cellular marrow admixed with individual clusters of Gaucher cells (PAS+, CD68+, Fe+) Accumulation of glucocerebroside (a lipid) in histiocytes (spleen, LNs, BM), Kupffer cells (liver), osteoclasts (bone), microglia (CNS), alveolar macrophages (lung)

Answer 70

_Type I (nonneuronopathic, adult)_: 90% of cases; first appears in adult liufe with hepatosplenomegaly, bone pain, pathologic fx, or pancytopenia; no CNS involvement; dx by Gaucher cells on bx and enzymatic assays indicating reduced but detectable levels of gluocerebrosidase activity; life not markedly shortened _Type II (acute neuronopathic,infantile)_: infancy and assd with CNS dysfunction, including convulsions and progressive mental deterioration; no detectable glucocerebrosidase activity; die by age 2 _Type III (chronic/subacute neuronopathic, juvenile)_: intermediate b/w I and II; presents in adolescents; slowly progressive; small amnt of glucocerebrosidase activity may be detectable

Answer 71

- AR lysosomal storage dz - in types A and B, deficiency of sphingomyelinase casues accumulation of sphingomyelin, a fatty substance present in every cell of body - characteristic cell is a lipid-laden macrophage referred to as "foam cell;" cytoplasmic inclusions are relatively uniformly-sized globules - may see vacuolated lymphocytes and monocytes in PB

Answer 72

``` _Type A (most common)_: occurs in infants and is characterized by jaundice, hepatomegaly, profound brain damage; children with this type rarely live beyond 18 months _Type B_: usually occurs in pre-teen years; pts have hepatosplenomegaly but no brain involvement _Types C and D_: may appear in early life or develop in teen or adult years; moderate splenomegaly, but brain damage may also be extensive; these type characterized by defect that disrupts transport of ***cholesterol*** b/w brain cells ```

Answer 73

Amino acid substitutions that affect: 1) binding of globin chains to heme OR 2) points of contact b/w globin chains - affect protein stability and result in intracellular ppt of globin chains - ppt globin chains attach to RBC membrane, giving rise to Heinz bodies - Hg Koln is the most common variant; others include Hb Christchurch and Hb Sydney - HbA will behave like unstable Hb when subjected to proportionately greater heat stress - Heinz body positive hemolytic anemia (due to unstable Hb, NOT G6PD def) the heat stability and isopropanol tests can be used to confirm presence of an unstable Hb variant

Answer 74

_Heat Stability Test_ _Isopropanol Stability Test_

Answer 75

- when Hb is heated, stability of molecule decreases; under controlled conditions, unstable Hb ppt, whereas stable Hb remain in solution - method: hemolysate is added to a Tris-HCl buffer and heated in water bath at 50 deg Celsius; tube is examined at 60, 90, and 120 minutes for ppt

Answer 76

- when Hb is dissolved in isopropanol, which is more nonpolar than water, the hydrophobic bonds are weakend and stability of molecule decreases - method: hemolysate added to a 17% isopropanol buffer solution and heated in water bath at 37 deg Celcius; tube is examined at 5, 20, 30 min for ppt \*samples with increased HbF (5-10% or more) will produce a positive test; methemoglobinemia (usually due to prolonged storage) will also give a positive result\* -false negatives are avoided by continuing the incubation until a control sample undergoes ppt

Answer 77

- acquired clonal genetic disorder that results in chronic intravascular hemolysis - acquired mutation in the phosphatidylinositol glycan A (PIG-A) gene on the X chromosome - PIG-A gene responsible for synthesis of glycosylphosphaidylinositol (GPI), which anchors certain proteins to the cell membrane; b/c GPI linked proteins inactivate complement, their absence renders affected blood cells unusually sensitive to lysis by complement - mutation occurs in a pluripotent stemm cell (thus RBCs, WBCs, and plts will be deficient in GPI proteins - most pts have a large pop of unaffected cells b/c the mutation affects only the clonal progeny of a single pluripotent stem cell

Answer 78

_RBCs_: CD55 (DAF), CD59 (MIRL, potent inhibitor of C3 convertase that normally prevents spontaneous activation of complement) _Monos_: CD14 _Grans_: CD24,CD66b,CD55,CD59

Answer 79

- plays the major role in development of hemolysis - CD59 neutralizes cleaved portion of C5 - when CD59 is absent, cleaved part of C5 causes cell lysis

Answer 80

- 50% of aplastic anemia pts have expansion of PNH clones - ~5% of aplastic anemia pts develop PNH - ~25% of pts receiving antithymocyte serum for the treatment of AA recover with evidece of PNH - the somatic PIG-A gene mutation is relatively common; a PNH clone emerges b/c the normal cells are suppressed (by antithymocyte serum or by a native imunological attack)

Answer 81

- begins as insidious onset of anemia - chronic hemosiderinuria is a constant feature and may result in Fe deficiency anemia - thromboses (due to abnormal plt function) are most common cause of morbidity/mortality - increased risk for AML and MDS - tx with ***eculizumab*** (monoclonal Ab to C5, prevents it from being cleaved into active form) - cure with BMT

Answer 82

_CBC_: anemia, leukopenia, thrombocytopenia (may also occur b/c overall deficiency in hematopoiesis) _Chem_: increased LDH (from RBC hemolysis), hemosiderinuria _Flow_: of PB is the gold standard for dx; defect is often more apparent in grans and monos vs RBCs (b/c RBCs are more sensitive to complement-mediated lysis than leukocytes, often diluted by transfused RBCs) FLAER: (fluorescent aerolysin, derived from from *Aeromonas*). Aerolysin binds GPI part of GPI-linked molecules on the cell surface (ie bind to normal cells but not PNH cells) _Positive sucrose hemolysis test (screening test)_: RBCS are incubated in serum and isotonic sucrose, which promotes complement binding; after incubation for 60 min, \>5-10% hemolysis is a positive result _Positive acified serum lysis test (Ham's test)_: acidifaction of serum activates complement leading to lysis of PNH cells

Answer 83

- erythroblasts mature into marrrow reticulocytes in about 6 days - each erythroblast can give rise to ~8 reticulocytes

Answer 84

Only a few hours before moving into tissues

Answer 85

Impedence: cell vol/size Conductivity: cell complexity Light scatter: cytoplasmic granularity light scattter (x-axis) vs impedence (y-axis) creates the WBC histogram (axes are opposite of flow)

Answer 86

ITP + hemolytic anemia

Answer 87

~2000-4000

Answer 88

- accounts for 98% of anemia in children less than 4yo; in adults due to blood loss - in mild Fe def anemia, decreased serum iron levels and increased TIBC usually precede changes in RBC morphology or CBC indices - pts may have cheilitis, koilonychia (spoon nails), pica

Answer 89

decrease ferritin→decrease transferrin sat→decrease in serum Fe→increase in ZPP→decrease in Hb, normocytic, normochromic→microcytic, hypochromic anemia

Answer 90

- microcytic, hypochromic anemia w/ high RDW - low ferritin - low serum iron - high TIBC - low transferrin sat (often less than 10%) - high serum transferrin - high serum soluble transferrin receptor - high free erythrocyte protoporphyrin - absent marrow iron stores - low serum hepcidin

Answer 91

- defective iron cycling b/w macrophages and erythroid precursors, iron becomes trapped within ***macrophages*** and is unavailable for heme synthesis - chronic inflammatory d/o (SLE, RA, IBD, sarcoidosis), chronic infections (HIV, TB, pyelo, osteo, chronic fungal infx, SBE), neoplasms, and end organ failure (chronic liver dz, endocrinopathies) - anemia develops ***1-2 months*** after onset of dz - immunologic response to protect host from invading organisms/developing neoplasms by depriving of iron

Answer 92

- normocytic normochromic - normal-sl increased RDW - normal or high ferritin - low serum iron - low TIBC - low transferrin sat - normal-sl increased serum soluble transferrin receptor - high free erythrocyte protoporphyrin - marrow iron stores increased in macrophages/reticuloendothelial cells, but decreased in sideroblasts

Answer 93

- Fe phsyiology and heme synthesis are normal; defect is globin synthesis - microcytic, hypochromic anemia - normal RDW - normal to increased RBC number - often see basophilic stippling - normal-high serum Fe - normal-low TIBC - normal-high transferrin sat - variable serum soluble transferrin receptor (may be high) - marrow iron stores are increased (due to ineffective erythropoiesis)

Answer 94

MCV/RBC less than 13: thalassemia greater than 13: Fe deficiency

Answer 95

- iron overload state characterized by abnormal iron metabolism and heme synthesis within RBC - Fe is sequestered in RBC ***mitochondria***, making it unavailable for heme synthesis (mitochondria typically perinuclear-hence ring sideroblasts) - may be hereditary (either X-linked or autosomal) or acquired later in life (as part of MDS or 2/2 toxic insult like drugs, EtOH, lead) - *hereditary* forms may respond to tx with pyridoxine

Answer 96

- basophilic stippling on PB - classically dimorphic pop of macrocytic cells and normocytic/microcytic cells- particularly in acquired forms - high ferritin - high serum iron - normal-low TIBC - high transferrin sat - variable serum soluble transferrin receptor (may be high) - increased marrow iron stores

Answer 97

- hereditary neutropenia (along with Kostmann syndrome) - AD, some sporadic - mutation in neutrophil elastase gene (ELA2) - neutrophil elastase is a granule serine protease - 21 day oscillating PMN count with ANC 0-1.5 x10⁹/L (severe infx at low level) - PB monos trend opposite ANC (high when ANC low) - respond to G-CSF (shortens cycle length and increase amplitude of waves) - does NOT progress to MDS/AML

Answer 98

- marked absolute neutropenia, usually less than 200/µL - increased susceptibility to bacterial infections - true Kostmann Syndrome is AR (other forms of SCN can be AD/sporadic) - 60-90% cases have point mutation in neutrophil elastase gene (ELA2)- same gene but different mutations as cyclic neutropenia - distinguish from Schwachman-Diamond Syndrome by lack of exocrine pancreas deficiency and growth retardation - ***promyelocytic/myelocytic maturation arrest in BM*** - increased monos and eos in BM (alternate fates of progenitor cell) - 90% of pts respond to G-CSF (no toxic type granulation-weird) - MDS/AML in 13%, cumulative risk with 8yrs G-CSF - cure with BMT

Answer 99

Non-specific, azurophilic - appear "late" in myeloblasts and promyelocytes, often remain visible in myelocytes - coated with phospholipid membrane - contain hydrolytic enzymes (MPO, lysozyme, acid phosphatase) - visible in mature neutrophils as "toxic granulation"

Answer 100

Specific - appear in myelocytes - like primary granules, secondary lysosomal granules contain numerous cytolytic enzymes (leukocyte alkaline phosphatase)

Answer 101

\*In reactive monocytosis, immature forms (ie monoblasts and promonocytes) are not seen\* - Infection (TB, listeria, syphilis, some protozoal/rickettsial infections) - Neoplasms (lymphoma, carcinoma, myeloma) - Miscellaneous (post-splenectomy, HA, ITP) - Inflammatory d/o (IBD, collagen vascular dz, sarcoid, celiac) - Chronic neutropenia (during recovery phase from acquired or cyclic neutropenia)

Answer 102

- third type of granule subset - recently identified in band and segmented neutrophils

Answer 103

- congenital d/o characterized by severe neutropenia - chacteristic findings include degenerative changes and hypersegmentation of mature neutrophils and hyperplasia of BM myeloid cells (PMNS can have bisegmented nuclei connected by long filaments leading to bizarre 'eyeglasses' forms) - recurrent bacterial infections (2/2 neutropenia AND depressed PMN fucntional activity) - patholphysiology due to prolonged retention of PMNs in BM compartment (kathexis=retention) - partially corrected by G-CSF

Answer 104

- warts, hypogammaglobulinemia, infections, myelokathexis syndrome - rare congenital immunodeficiency characterized by susceptibility to HPV-induced warts and carcinoma, neutropenia, B-cell lymphopenia and hypogammglobulinemia related infections, and BM myelokathexis - caused by mutations in gene coding for chemokine receptor CXCR4, leading to mutant protein that causes abnormal apoptosis and migratory fx of leukocytes

Answer 105

- less than 2.5 x 10⁹/L in infants - less than 1.5 x 109/L in kids and adults - "severe" is less than 0.5 x 10⁹/L

Answer 106

- infection - maternal hypertension or drugs - maternal ab production - constitutional d/o (cyclic neutropenia, Kostmann syndrome, Chediak-Higashi, Schwachman-Diamond, myelokathexis, Fanconi anemia, dyskeratosis congenita)

Answer 107

- infection - autoimmune neutropenia (BM shows macrophages with ingested PMNs) - myelopthisic disease - Idiosyncratic drug reactions - 2/2 autoimmune neutropenia in collagen vascular d/o - immunodeficiency - myeloablative tx

Answer 108

- infection - idiosyncratic drug rxn - myelophthisic dz - myeloablative tx - secondary autoimmune neutropenia in collagen vascular d/o - autoimmune d/o - aplastic anemia - immunodeficiency - hypersplenism (due to sequestration) - megloblastic anemia - Felty syndrome (triad: RA, splenomegaly, neutropenia)

Answer 109

- RA - splenomegaly - neutropenia (risk for infections) - affects women, 50-70 yo

Answer 110

- rare, often fatal d/o of infancty - due to large deletion of mitochondrial DNA (maternally inherited) - deletions of certain components of electron transport chain cause defect in cellular oxidative metabolism; also impaired traslation of mRNA - diagnose by detection DNA deletion via Southern blot

Answer 111

- severe, transfusion dependent macrocytic anemia begins early in infancy - ring sideroblasts and vacuolated marrow precursors are variably present - variable degree of neutropenia/thrombocytopenia - lactic acidosis - exocrine pancreatic insufficiency and often renal dysfunction - progressive dz - pts who survive infancy→ ***Kearns-Sayre syndrome***: * progressive external ophthalmoplegia * weakness of skeletal muscles * multisystemic d/o

Answer 112

cell with visible Golgi likely to be myelocyte than promyelocyte

Answer 113

scanty agranular cytoplasm, medium sized nucleus wtih delicate, evenly distributed chromatin, and 1 or more inconspicuous nucleoli

Answer 114

larger cells with relatively abundant cytoplasm and prominent large nucleoli

Answer 115

on 100x, number of plts x 13,000

Answer 116

Cells usually larger tahn normoblastic counterparts 1. Megaloblastic type: - N:C dyssynchrony, can be seen with hydrea therapy 2. Megaloblastoid type: - more subtle, most prominent feature is abnormal chromatin pattern that is more condensed and clumped, with more prominent parachromatin spaces, also mild N:C dyssynchrony

Answer 117

monolobated nucleus with pink cytoplasm

Answer 118

- represent pre-pre-B cells and pre-B cells and are not seen in blood - seen post chemo (recovering BM) and in children - cells have condensed nucledar chromatin and very scant cytoplasm - similar to B lymphoblasts on flow (positive for CD10, CD19, CD34, TdT; negative for surface light chain) - on BM biopsy, small clusters of 5-6 cells

Answer 119

-small with immature (blue) cytoplasm and no granules

Answer 120

small with mature (pink) cytoplasm

Answer 121

dense, dark purple granules cover the cell and obscure the cytoplasmic edge of cell

Answer 122

- large multinucleated cells with multiple oval nuclei - in contrast to megas, the nuclei are completely separate from one another

Answer 123

- associated with ertain infections (EBV), autoimmune dz, some malignancies (T cell lymphomas) - internalized cells are visibly degenerated, in contrast to emperipolesis in which internalized cells are intact

Answer 124

- monos are the first cell type to recover following BM insults (chemotherapy, aplastic anemia) - other causes of monocytosis include chronic infection (listeria), collagen vascular diseases, and CMML

Answer 125

- seen in MPNs (especially CML) - storage diseases (Niemann-Pick dz, Fabry dz) - occasionally seen in normal marrows

Answer 126

- maturation occurs predominantly along bony trabeculae - Day 14: marrow should be 0% cellular (may have some plasma cells); if greater than 5% by ***morphology*** (not flow), then clinician will try to reinduce remission - order of BM recovery after transplant: erythroid, megas, myeloid - avg of 1-3 megas/50x field is normal

Answer 127

interstitial: more likely B9 or CLL paratrabecular: follicular lymphoma diffuse B cells: hairy cell

Answer 128

may be seen in HIV infection

Answer 129

- clear cytoplasm in erythroid - pink cytoplasm in myeloid and plasma cells

Answer 130

-larger, blast like B cells with vesicular nuclei with fine chromatin and 2-4 membrane bound nucleoli (dark zone within germinal centers)

Answer 131

-cells with large central nucleoli, appreciable amount of basophilic cytoplasm, CD30+, CD20+, BCL2+

Answer 132

-same as prolymphocytes (pale pseudofollicles in CLL/SLL)

Answer 133

-small B cells with irregular/cleaved nuclear contours and scant cytoplasm (light zone within germinal centers)

Answer 134

-large, round, vesicular nuceli with open chromatin, one central nucleolus, often occur in pairs (like R-S cells)

Answer 135

- follicular hyperplasia - follicular lymphoma - HIV - RA and Sjogren's syndrome lymphadenitis - syphilis - Castleman's dz - PTGC

Answer 136

-reactive follicular hyperplasia with interfollicular plasmacytosis

Answer 137

-similar to RA lymphadenitis, but also showing capsular and trabecular thickening with infiltration by plasma cells

Answer 138

- usually affects mediastinal nodes - plasma cell variant associated with increased IL-6

Answer 139

- viral infection (EBV-infectious mono, CMV, postvaccinial lymphadenitis) - hypersensitivity reaction (dilantin) - Kimura's dz: * young asian men, cervical lymphadenopathy * florid follicular hyperplasia with interfollicular immunoblasts, increased vascularity, increased eos in paracortex, sinusoids, perinodal soft tissue, and within follicles

Answer 140

- sinus histiocytosis - Rosai-Dorfman disease - Whipple dz (M:F = 10:1) - dermatopathic change - hemophagocytic syndrome - vascular transformation of LN sinuses (abdominal nodes)

Answer 141

-suppurative granulomas: * cat scratch (stellate abscess) * lymphogranuloma venereum * tularemia -necrotizing granulomas: * TB/atypical mycobacteria * Brucellosis * fungal infection * Yersinia - non-necrotizing granulomas: * sarcoid - granulomas impinging on germinal centers: * toxoplasmosis (triad: reactive follicular hyperplasia, epithelioid histiocytes encroaching germinal centers, monocytoid B cell hyperplasia) - ill defined sheets of histiocytes with karyorrhexis: * Kikuchi-Fujimoto

Answer 142

- EBV (may be interfollicular; node effacted by proliferation of T cells, plasma cells, immunoblasts that may be very atypical and mimic HL or large cell lymphoma) - CMV (see typical inclusions) - HSV (focal necrosis is characteristic, typical inclusions may be seen) - measles (Warthin-Finkeldy giant cells) - post-vaccinia

Answer 143

-CD41, CD42b, CD61

Answer 144

- plasma cells - immunoblasts

Answer 145

- T cell marker - surrogate marker for clonality in B cells

Answer 146

T cell marker

Answer 147

immunoblasts

Answer 148

- initial method used; pH 8.6 - all Hb variants at this pH are negatively charged and migrate to anode - starting at anode: A Fat Santa Claus - band in S region: Hb S (solubility test +), Hb G, Hb D, and Hb Lepore (solubility test -) - band in A2 region: C, E, O-Arab

Answer 149

- pH 6.2 - starting at cathode: Fat Ass Santa Claus - useful to separate bands at S and A2 on cellulose agar - variants that co-migrate with Hb A: A2, D, G, E, O-Arab

Answer 150

-very mild microcytosis (if less than 75, consider concomitant thal), scattered target cells, pts asymptomatic

Answer 151

-mild hemolytic anemia, mild splenomegaly (pts mostly asymptomatic), mild microcytosis, numerous target cells, hexogonal or rod shaped crystals in RBCs in splenectomized pts

Answer 152

- sickling dz milder than SS - true sickle cells and Hb C crystalss are rare; there are usually irregular shaped cells that contain misshapen crystals

Answer 153

-solubility test is positive

Answer 154

-mild microcytosis, scattered target cells, no anemia

Answer 155

-MCV around 67, frequent target cells, mild or no anemia

Answer 156

- SE Asians - target cells, thalassemic indices - mutation causing beta globin mRNA to be underexpressed and is functionally more like beta thal - migrates with C, A2, and O on alkaline (AKA cellulose acetate) - elutes with A2 on HPLC

Answer 157

- clinically insignificant - AKA HbD-Los Angeles or HbD-Punjab - combo of HbD and HbS produces severe sickling disorder

Answer 158

- most common alpha globin gene mutation seen in US (mainly AA) - clinically normal

Answer 159

- heterozygotes: no hematologic abornmalities - homozygotes: mild anemia - with HbS: produces severe sickling d/o similar to SS

Answer 160

- fusion of delta and beta genes during meiotic crossover (3 different forms exist) - stable hemoglobins that are underproduced relative to normal beta chains, thus Hb Lepore gene functions as a thalassemic allele - Mediterranean pts - a minor band in the HbS position (10-15%) on alkaline (celluose acetate), negative solubility test, coelution with HbA2 on HPLC is virtually diagnostic of Lepore - mild microcytosis and occasional target cells

Answer 161

- produces thalassemic indices - results from mutation of the alpha stop codon, producing an abnormally long transcript that is unstable (and functionally more like thalassemia than hemoglobinopathy)

Answer 162

- rare; unable to maintain heme iron in reduced state or to reduce 02 - results in hereditary methemoglobinemia

Answer 163

elavated HbF into adulthood (20-40%) _deletional mutations_: - involve large deletions spanning the gamma and beta genes on chromosome 11 - MCV is normal-sl decreased, no anemia - pancellular pattern on K-B acid elution test (all RBCs contain similar amount of HbF) or by flow cytometry using an anti-HbF ab _non-deletional mutations_: - typically due to point mutations in the promoter region of one of the gamma globin genes, resulting in persistent epression of HbF (the gamma globin switch is not turned off in adulthood) - pancellular and heterocellular pattern on K-B test or by flow - measurement of individual gamma chains usually show abnormal gamma G to gamma A ratio