Are Tidal Salt Marshes Exposed to Nutrient Pollution more Vulnerable to Sea Level Rise?

Over the past four decades, Long Island, NY, USA, has lost coastal wetlands at a rate of 4% per decade due to submergence. In this study, we examined relationships between the rate of tidal salt marsh loss and environmental factors, including marsh elevation, tidal range, and wastewater exposure through analysis of stable isotope ratios of marsh soils and biota. Our goal was to identify factors that increase vulnerability of marshes to sea level rise, with a specific emphasis on the potential role of poor water quality in hastening marsh loss. Our results suggest that wastewater exposure may accelerate loss of intertidal marsh, but does not negatively impact high tidal marsh resilience to sea level rise. And while marsh elevation and tidal range were statistically significant predictors of marsh loss, they similarly displayed opposite relationships among marsh zones. This study suggests that different functional zones of coastal salt marshes may not respond similarly to global change factors, and that elevation may be an important factor mediating eutrophication effects to coastal salt marshes.

Granulocytopenia in cancer patients treated in a phase I trial with recombinant human tumor necrosis factor.

We have examined the effect of recombinant human tumor necrosis factor (TNF) upon granulocyte kinetics in cancer patients in a phase I clinical trial. TNF was given to each patient intravenously over 2 h at varying doses. A marked drop in the total white blood cell count, absolute polymorphonuclear leukocyte (PMN) count, and absolute monocyte count occurred reproducibly at 30 min after TNF initiation. Also noted was a drop in the absolute lymphocyte and eosinophil counts, both of which reached their nadir at approximately 4 h. A marked increase in immature PMN leukocytes (bands) was noted beginning at 1 h. These changes were statistically significant. Statistically significant increases in hemoglobin and hematocrit occurred at the 30 min time point but subsequently decreased to approximately 90% of pretreatment baseline. Additionally, the platelet count decreased, reaching its nadir approximately 6 h after TNF initiation. In four serial studies in patients on the highest dose of TNF, the granulocyte adhesion protein CD11b was shown to increase on the surface of the PMN leukocytes by as early as 7-15 min after initiation of TNF infusion. In each of these, expression of CD11b antigen increased prior to the disappearance of PMN leukocytes from the peripheral circulation. A similar finding was obtained for monocytes. This work indicates that within 30 min of intravenous infusion of TNF, mature granulocytes and monocytes have left the peripheral circulation. This is followed by an apparent bone marrow response indicated by an outpouring of bands. The initial granulocyte and monocyte emigration from the peripheral circulation is preceded at highest-dose TNF by increased cell surface expression of CD11b for both cell types, suggesting a causal relationship between these temporally linked events.

Morphometric study of megakaryocytes size and prognosis in adults with acute non-lymphocytic leukemia.

Morphometric analysis of megakaryocyte cellular and nuclear size in bone marrow biopsies of 28 cases of acute non-lymphocytic leukemia at diagnosis and 15 controls was performed. Median megakaryocyte cell diameter, median area, and perimeter were less than 95% control range (less than 18.5 microns, less than 263 microns2, less than 64 microns) in 5/5 of induction failures. Attainment of complete remission was significantly greater in those with median megakaryocyte diameter greater than or equal to 18.5 microns (p less than 0.01), median area greater than or equal to 263 microns (p less than 0.001) and perimeter greater than or equal to 64 microns (p less than 0.001). Prolonged complete remission was correlated with normal megakaryocyte size with the median megakaryocyte area, median diameter, and perimeter within or greater than the reference range (greater than or equal to 18.5 microns, greater than or equal to 263 microns2, greater than or equal to 64 microns, p less than 0.05) in 6/7 cases with continuous remissions greater than 3 yr. Measurements of megakaryocyte size may be useful in predicting induction failure and possibly the likelihood of prolonged complete remission in adults.

Acquired factor X deficiency and amyloidosis.

A selective acquired Factor X deficiency is an unusual occurrence. Six cases of an acquired Factor X deficiency in association with amyloidosis have been reported. This paper describes two additional cases, suggesting that this relationship may be more than coincidental. The mechanism by which amyloid may affect Factor X levels remains unknown, but suggestions include consumption, inactivation or decreased synthesis of Factor X. Factor II, VII, IX, and X concentrate transiently increased the Factor X level to normal in one of the patients. In an adult patient who has an isolated Factor X deficiency, amyloidosis should be actively sought.

Automated differentials in the hematology laboratory.

The white blood cell differential continues to be one of the most widely performed clinical laboratory procedures. However, its clinical usefulness is affected by sampling error and, to some extent, by classification criteria. It is also labor intensive and expensive to perform. Automated leukocyte differential instrumentation addresses many of the sources of error that occur with the manual differential. Current state-of-the-art instrumentation will give results that equal or exceed the routine manual differential. Because these instruments also examine red blood cell and platelet parameters, as well as providing white blood cell information, they can better screen for significant abnormalities as well as greatly reduce the expensive and time-consuming manual differential procedures.

Serum ferritin and bone marrow biopsy iron stores. II. Correlation with low serum iron and Fe/TIBC ratio less than 15%.

Diseases associated with defective reticuloendothelial release of iron may be difficult to distinguish from the iron-deficient state, since serum iron parameters (serum iron, percentage saturation of transferrin less then 15%) may overlap. Assessment of the bone marrow biopsy iron stores was often necessary to resolve the diagnosis. Serum ferritin levels have proved very useful in distinguishing the uncomplicated iron-deficient state from other disorders associated with low serum iron and low percentage saturation of transferrin. However, a small percentage of iron-deficient persons may have normal ferritin values, particularly if these persons have liver disease or hematopoietic or lymphoreticular neoplasms. In these cases, assessment of the bone marrow biopsy iron would still be necessary to evaluate the body iron stores.

Serum ferritin and bone marrow iron stores. I. Correlation with absence of iron in biopsy specimens.

Serum iron, total iron-binding capacity, and percentage saturation of transferrin have classically been used to demonstrate a hypoferremic state; however, these tests may not discriminate between depleted iron stores and conditions associated with defective reticuloendothelial release of iron. Estimation of stainable iron in the bone marrow biopsy specimen is then the most practical way to assess body iron stores. With the availability of a radioimmunoassay procedure for serum ferritin, we undertook a prospective study to determine whether serum ferritin concentrations might replace assessment of the marrow biopsy iron stores as an indicator of hypoferremia. Iron stores were absent from bone marrow biopsy specimens from 104 patients. A good correlation between low serum ferritin levels and absence of iron stores in biopsy specimens was found for 91 patients (87.5%). Thirteen (12.5%) had normal serum ferritin concentrations with absence of biopsy iron. These individuals had hematopoietic malignancies or active hepatic disease, or were receiving iron therapy. In this group, a bone marrow biopsy would still be necessary for evaluation of a hypoferremic state, even though the serum ferritin concentration might be normal.

The effect of hemoglobin F upon glycosylated hemoglobin determinations.

Glycosylated hemoglobin (Hgb A1) determinations have been advocated for monitoring the control of diabetes mellitus. The prevalent method today for measuring Hgb A1 for most clinical laboratories has been a "mini-column" utilizing ion exchange chromatography. It has been stated that hemoglobin F (Hgb F) will elute with Hgb A1 and interfere with Hb A1 determinations. This study was designed to determine the quantitative effects of Hgb F upon Hgb A1 determinations. Thirty per cent of the study group had elevated Hgb A1 levels at 2% Hgb F concentration, 66% at 3% Hgb F concentration, and all individuals had elevated Hgb A1 levels at Hgb F concentrations of 4% or greater. The relationship of Hgb F to Hgb A1 concentration was not a simple identity, but could be represented by the equation y = 6.03 + 1.24x. If the ion exchange chromatography methodology is used, Hgb F levels should be determined whenever Hgb A1 levels are elevated, particularly in populations where increased Hgb F levels also might be encountered. The authors determined Hgb F levels whenever the concentration of Hgb A1 was 10% or greater. In their population, they found that approximately 1.5% of samples with elevated Hgb A1 concentrations had increased (greater than 2%) Hgb F levels.

Comparison of stainable iron in aspirated and needle-biopsy specimens of bone marrow.

A comparison of stainable iron in simultaneously obtained aspirated smears and needle-biopsy specimens from 1,000 patients was undertaken. Significant differences occurred when iron was assessed as absent in the aspirated smear. In only 35% of the corresponding needle-biopsy specimens was iron absent. When only the aspirated smear was used, there was a significant overdiagnosis of iron deficiency. In general, iron tended to be less in the aspirated smear; however, correlations were better when iron stores were assessed as being present or increased in the aspirated smears, for stainable iron in the needle-biopsy specimen was always present in equal or greater amounts. Hemosiderotic smears (increased stainable iron) and needle-biopsy specimens (3+ -4+) correlated well. The aspirated smear and needle-biopsy are complementary procedures, and each has advantages. In the authors' experience, the needle-biopsy was preferable to the aspirated smear for evaluation of iron stores, particularly when iron stores were low or absent.

Use of the bone marrow imprint in the diagnosis of leukemic reticuloendotheliosis ("hairy cell leukemia").

Leukemic reticuloendotheliosis is a distinct entity, often misdiagnosed as chronic lymphocytic leukemia or lymphoma. The neoplastic cells have a specific tartrate-resistant acid phosphatase isoenzyme by which the diagnosis may be secured. Most individuals are leukopenic, and when, as in our case, rare or no circulating cells with tartrate-resistant acid phosphatase activity are found in the peripheral blood, an alternate site must be sought. Bone marrow aspirations often result in "dry taps," however, and cryostat sections of the bone marrow biopsy necessary to demonstrate tartrate-resistant acid phosphatase activity are involved and impractical to obtain for most laboratories. This report illustrates and recommends the simple technic of imprinting the core biopsy, which yields a satisfactory sample with which the specific cytochemical activity can be demonstrated.

Flow cytometry in the diagnosis of acute leukemia.

Fifty cases of acute leukemia were analyzed by means of flow cytometry. The results obtained were correlated with morphology and routine cytochemistries. The panel selected was useful in classifying an acute leukemia as acute lymphocytic (ALL) or acute nonlymphocytic (ANLL), which is of primary importance for therapeutic considerations. Common ALL (CALLA) (J5) was a good marker for classifying the leukemia as ALL. Monoclonal antibodies (MoAbs) T1 and/or T11 further delineated the lymphoid leukemia as T-cell ALL while MoAbs B4 or B1 delineated the lymphoid leukemia as non-T-cell ALL. Eighteen cases of ALL were diagnosed and consisted of five cases of T-cell ALL and 13 cases of non-T-cell ALL. Both the T-cell ALL cases and non-T-cell ALL cases were found to be heterogeneous and could be further subgrouped by phenotypic expression with additional MoAbs in the panel. A monoclonal antibody panel consisting of My4, My7, My9, Mo1, and Mo2 was useful in characterizing an acute leukemia as ANLL. This panel was less useful in distinguishing myeloid from monocytic subtypes although My4, Mo1, or Mo2 when present, appeared to favor a monocytic component. Of interest, a case of biclonal leukemia with two distinct blast populations on the flow cytogram was discovered. Morphology alone was successful in diagnosing ALL from ANLL in 35 cases (70%). It was not useful in distinguishing non-T-ALL cases from T-ALL cases. The ambiguous cases could be resolved by cytometric means. Flow cytometry has much to offer as a diagnostic aid in the evaluation of acute leukemia.

Quality control in flow cytometry for diagnostic pathology. I. Cell surface phenotyping and general laboratory procedures.

Flow cytometric analyses have become commonplace in the clinical laboratory for determining cell lineage and for quantitation of cells bearing a given phenotype. Because these assays are being conducted to support diagnoses or assist in determining therapy, it is crucial to ensure that these tests are highly accurate and reproducible within a laboratory and among laboratories involved in similar endeavors. This quality assurance has been slow evolving in clinical flow cytometry for a variety of reasons: the exquisite sensitivity and delicacy of the instrumentation that recognize previously undetectable variations in staining; the constant improvement of the hardware and software; the rapid development of new techniques and reagents of clinical interest; and the failure of any existing specialty or subspecialty to encompass all aspects of flow cytometry. This article provides an overview of quality assurance necessary for the flow cytometric analysis of cell surface markers. Practical experience, published studies, and suggested guidelines from accreditation agencies have been combined to develop the text.

Leukemia with multiple phenotypic expressions.

A patient is described who, during a three-year illness, exhibited multiple phenotypic expressions of leukemia. She was diagnosed initially as having acute lymphoblastic leukemia (ALL) followed by chronic myelogenous leukemia (CML) one and a half years later. Subsequent blast transformations were lymphoblastic, but preterminally, a myeloblastic transformation occurred. These later leukemias probably were not chemotherapy induced, because a review of her initial work-up revealed the presence of a smaller G group chromosome in 1 of 20 metaphases examined. Her clinical course also differs from the CML that presents in lymphoid blast crisis because the Philadelphia chromosome only was emerging at that point. This case illustrates the interrelationships of the various leukemias and supports the hypothesis that a totipotent stem cell may undergo leukemic transformation resulting in variable and simultaneous expressions.

Diagnostic value of DNA analysis in effusions by flow cytometry and image analysis. A prospective study on 102 patients as compared with cytologic examination.

One hundred twenty-six effusion samples from 102 patients were examined by cytology and flow cytometry (FCM). Overall, there was an 84% correlation between cytologic and FCM results. Of the 36 malignant cases determined by cytologic examination, FCM revealed an aneuploid peak in 20 (56%). Image analysis (IA) performed on the malignant cytologic cases with a diploid flow pattern detected two additional aneuploid peaks. In addition, FCM indicated three aneuploid cases in which cytologic characteristics were initially interpreted as benign (false negative). Aneuploidy was therefore detected in 64% of the malignant effusion specimens by FCM and IA. Twenty-three of the total of 24 aneuploid cases detected by FCM were associated with malignancy (predictive value = 96%). The one nonmalignant case was that of hemorrhagic pancreatitis with infected pseudocyst. FCM is an excellent tool when moderate to large numbers of tumor cells are present, whereas use of IA is advantageous for specimens containing smaller numbers of malignant cells because these can be directly analyzed. When an aneuploid peak is present, a diagnosis of malignancy must be suspected, and, if the initial cytologic screen is negative, a critical review of the cytology slides is justified. In those cases with an equivocal atypical cytology report and an abnormal cytometric histogram, additional investigation is warranted. In some malignancies the tumor cells will be diploid (in this study 36%) and neither FCM nor IA will add to tumor detection, leaving cytologic examination as the definitive technique.

Simultaneous evaluation of terminal deoxynucleotidyl transferase and myeloperoxidase in acute leukemias using an immunocytochemical method.

The classification of acute leukemia is important for the selection of optimal therapy. Classification often rests on morphologic, cytochemical, and immunologic criteria, and the marker enzyme terminal deoxynucleotidyl transferase (TdT) has been considered to be a reliable indicator of lymphoblastic leukemias. Because TdT-positive cells sometimes are seen in leukemias otherwise identified as myeloblastic, the authors evaluated blasts identified as myeloid by the presence of myeloperoxidase (MPO) for the simultaneous expression of TdT. The blasts in the bone marrow aspirate or peripheral blood of unselected patients with hematologic malignancies were evaluated and 60 cases are shown. The French-American-British system and, in some patients, cytochemical and immunologic studies were used to classify the leukemias. The authors demonstrated that blasts simultaneously contained MPO and TdT in 29% of patients with acute myeloblastic leukemia and 3% of patients with acute lymphocytic leukemia (ALL). This finding supports the hypothesis that TdT is an expression of cell primitivity rather than a marker for lymphoblastic cells.

Detection of hepatitis C virus RNA sequences in B-cell non-Hodgkin lymphoma.

Serologic testing shows that hepatitis C virus (HCV) may have a role in the pathogenesis of B-cell non-Hodgkin lymphomas (B-cell NHLs). We tried to demonstrate HCV RNA sequences in paraffin-embedded tissue from B-cell NHLs by reverse-transcription double polymerase chain reaction (RT-PCR) and Southern blotting. We studied 31 consecutive cases of B-cell NHLs; lymph nodes from 32 patients with diseases other than B-cell NHL were negative controls. Positive-strand HCV RNA was tested with primers for the 5' untranslated region. Replicative negative strand HCV RNA was tested with strand-specific RT-PCR for the 5' untranslated region. Immunohistochemical staining for HCV was done using an antibody to HCV core protein. Positive-strand HCV RNA was detected in 8 patients with B-cell NHL; negative-strand HCV RNA was detected in 6 of these cases, indicating viral replication. All control cases were negative for HCV RNA. Immunohistochemistry showed no staining of lymphoma cells for HCV core proteins in any case. HCV and B-cell NHLs may be associated. RT-PCR on paraffin-embedded lymphoma tissue is an alternative method of testing for HCV. The value of immunohistochemistry could not be ascertained. The exact role of HCV in the pathogenesis of B-cell NHL needs to be studied further.

Effect of simulated altitude erythrocythemia in women on hemoglobin flow rate during exercise.

The effect of simulated altitude erythrocythemia on hemoglobin flow rate and maximal O2 uptake (VO2max) was determined for nine women sea-level residents. Test conditions included normoxia and normobaric hypoxia (16% O2-84% N2). Cycle tests were performed under normoxia (T1-N) and hypoxia (T1-H) at prereinfusion control and under hypoxia 48 h after a placebo infusion (T2-H) and 48 h after autologous infusion of 334 ml of erythrocytes (T3-H). Hematocrit (38.1-44.9%) and hemoglobin concentration (12.7-14.7 g.dl-1) increased from control to postreinfusion. At peak exercise, VO2max decreased from T1-N (2.40 l.min-1) to T1-H (2.15 l.min-1) then increased at T3-H (2.37 l.min-1). Maximal arterial-mixed venous O2 difference decreased from T1-N to T1-H and increased at T3-H. Cardiac output (Q), stroke volume, heart rate, and total peripheral resistance during maximal exercise were unchanged from T1-N through T3-H. Hemoglobin flow rate (Hb flow) at maximum did not change from T1-N to T1-H but increased at T3-H. When compared with submaximal values for T1-N, VO2 was unchanged at T1-H and T3-H; Q increased at T1-H and decreased at T3-H; arterial-mixed venous O2 difference decreased at T1-H and increased at T3-H; Hb flow did not change at T1-N but increased at T3-H. For young women, simulated altitude erythrocythemia increased peak Hb flow and decreased physiological altitude (227.8 m) but did not affect maximum cardiac output (Qmax).

A new procedure for cell lineage determination in acute leukemias. Myeloperoxidase mRNA detection.

Determination of cell lineage in acute leukemias is essential for diagnosis and treatment. Detection of myeloperoxidase (MPO) mRNA establishes myeloid lineage of leukemic blasts that may be too primitive to be identified as myeloblasts based on morphology, cytochemistry, or immunophenotype. A highly specific and sensitive new procedure for MPO mRNA detection has been developed using HL-60 cells. It involves a microprocedure for total cellular RNA extraction, reverse transcription, and specific amplification of target sequences in the resulting MPO cDNA, by the polymerase chain reaction. Specific primers are designed to amplify an 89-base pair (bp) sequence from the signal peptide, 179 and 318-bp sequences from the start and end, respectively, of the heavy-chain sequence, and a 255-bp sequence overlapping the proregion and light chain. The correct-size amplification products, detected electrophoretically, demonstrate MPO mRNA expression in the leukemic cells analyzed. The sensitivity of this new procedure was evaluated on serial concentrations of HL-60 cells and was found to be 10-10(4) cells depending on the MPO cDNA amplified sequence. No amplification products were obtained using peripheral blood lymphocytes as a negative cellular control. The specificity of the procedure is demonstrated by Southern blotting and hybridization with 32P-labeled oligonucleotide probes specific for each of the amplified sequences. An additional advantage of this procedure is availability of results in 8-24 h, compared with 1-2 weeks for conventional RNA methods.

der(1)t(1;19)(p12;p11): a new nonrandom chromosomal abnormality in myelodysplastic syndrome.

The der(1)t(1;19)(p12;p11) has not been previously reported in myelodysplastic syndrome (MDS). Fluorescence in situ hybridization (FISH) using chromosome 1- and chromosome 19-specific probes, performed on the bone marrow (BM) cells of this patient confirmed the initial karyotype, i.e., 47,XY,+der(1)t(1;19)(p12;p11).

Atypical (7;19) translocation in acute myelomonocytic leukemia.

Chromosome studies were carried out after a 24-hour harvest of unstimulated bone marrow aspirate cell cultures from a 75-year-old male with a clinical diagnosis of acute myelomonocytic leukemia (FAB M4). Analysis of nine cells after trypsin-Giemsa banding (GTG) revealed two cell lines with a mosaic chromosome pattern, 46,XY/46,XY,t(7;19)(q22;p13.3). A review of the recent literature reveals one case of childhood ALL with a 46,XY/46,XY,t(7;19)(q11;q13) chromosome pattern [1] and a 46,XY,t(3q;11q),t(7q;19p),t(15;17)(q26;q22) in one patient with ANLL (FAB M3) [2]. The t(7;19)(q22;p13.3) seen in our case has not been reported as the sole specific clonal chromosome rearrangement in myeloid neoplasia. Interestingly, the plasminogen activator inhibitor type I, multi-drug resistance, and erythropoietin genes are located at band 7q22 and the insulin receptor gene is located at band 19p13.3. Both sites contain fragile site loci. The possible role of these fragile sites, genes, or other genes in the rearrangement can only be surmised.