Proposed criteria for classification of acute myeloid leukemia in dogs and cats

Blood and bone marrow smears from 49 dogs and cats, believed to have myeloproliferative disorders (MPD), were examined by a panel of 10 clinical pathologists to develop proposals for classification of acute myeloid leukemia (AML) in these species. French-American-British (FAB) group and National Cancer Institute (NCI) workshop definitions and criteria developed for classification of AML in humans were adapted. Major modifications entailed revision of definitions of blast cells as applied to the dog and cat, broadening the scope of leukemia classification, and making provisions for differentiating erythremic myelosis and undifferentiated MPD. A consensus cytomorphologic diagnosis was reached in 39 (79.6%) cases comprising 26 of AML, 10 of myelodysplastic syndrome (MDS), and 3 of acute lymphoblastic leukemia (ALL). Diagnostic concordance for these diseases varied from 60 to 81% (mean 73.3 +/- 7.1%) and interobserver agreement ranged from 51.3 to 84.6% (mean 73.1 +/- 9.3%). Various subtypes of AML identified included Ml, M2, M4, M5a, M5b, and M6. Acute undifferentiated leukemia (AUL) was recognized as a specific entity. M3 was not encountered, but this subclass was retained as a diagnostic possibility. The designations M6Er and MDS-Er were introduced where the suffix "Er" indicated preponderance of erythroid component. Chief hematologic abnormalities included circulating blast cells in 98% of the cases, with 36.7% cases having >30% blast cells, and thrombocytopenia and anemia in approximately 86 to 88% of the cases. Bone marrow examination revealed panmyeloid dysplastic changes, particularly variable numbers of megaloblastoid rubriblasts and rubricytes in all AML subtypes and increased numbers of eosinophils in MDS. Cytochemical patterns of neutrophilic markers were evident in most cases of Ml and M2, while monocytic markers were primarily seen in M5a and M5b cases. It is proposed that well-prepared, Romanowsky-stained blood and bone marrow smears should be examined to determine blast cell types and percentages for cytomorphologic diagnosis of AML. Carefully selected areas of stained films presenting adequate cellular details should be used to count a minimum of 200 cells. In cases with borderline diagnosis, at least 500 cells should be counted. The identity of blast cells should be ascertained using appropriate cytochemical markers of neutrophilic, monocytic, and megakaryocytic differentiation. A blast cell count of > 30% in blood and/or bone marrow indicates AML or AUL, while a count of < 30% blasts in bone marrow suggests MDS, chronic myeloid leukemias, or even a leukemoid reaction. Myeloblasts, monoblasts, and megakaryoblasts comprise the blast cell count. The FAB approach with additional criteria should be used to distinguish AUL and various subtypes of AML (Ml to M7 and M6Er) and to differentiate MDS, MDS-ER, chronic myeloid leukemias, and leukemoid reaction. Bone marrow core biopsy and electron microscopy may be required to confirm the specific diagnosis. Immunophenotyping with lineage specific antibodies is in its infancy in veterinary medicine. Development of this technique is encouraged to establish an undisputed identity of blast cells. Validity of the proposed criteria needs to be substantiated in large prospective and retrospective studies. Similarly, clinical relevance of cytomorphologic, cytochemical, and immunophenotypic characterizations of AML in dogs and cats remains to be determined.     

Ultrastructural analysis of platelets and megakaryocytes from a dog with probable essential thrombocythemia

Blood platelets and bone marrow megakaryocytes from a dog with essential thrombocythemia were analyzed by transmission electron microscopy and compared with those of control dogs. Platelets varied in size and shape and contained enlarged and dilated open canalicular systems. Megakaryocytes were dysmorphic and had evidence of abnormal maturation, with large numbers of megakaryoblasts containing alpha granules and poorly organized and reduplicated demarcation membranes. The fewer, more mature megakaryocytes lacked normal cytoplasmic organization. Most notable was the absence of well-demarcated "platelet fields," due to the excessive and disorderly array of demarcation membranes.     

Ergovaline does not alter the severity of ryegrass staggers induced by lolitrem B

AIM: To investigate a possible interaction between lolitrem B and ergovaline by comparing the incidence and severity of ryegrass staggers in sheep grazing ryegrass (Lolium perenne) containing lolitrem B or ryegrass containing both lolitrem B and ergovaline.

METHODS: Ninety lambs, aged approximately 6 months, were grazed on plots of perennial ryegrass infected with either AR98 endophyte (containing lolitrem B), standard endophyte (containing lolitrem B and ergovaline) or no endophyte, for up to 42 days from 2 February 2010. Ten lambs were grazed on three replicate plots per cultivar. Herbage samples were collected for alkaloid analysis and lambs were scored for ryegrass staggers (scores from 0–5) weekly during the study. Any animal which was scored ≥4 was removed from the study.

RESULTS: Concentrations of lolitrem B did not differ between AR98 and standard endophyte-infected pastures during the study period (p=0.26), and ergovaline was present only in standard endophyte pastures. Ryegrass staggers was observed in sheep grazing both the AR98 and standard endophyte plots, with median scores increasing in the third week of the study. Prior to the end of the 42-day grazing period, 22 and 17 animals were removed from the standard endophyte and AR98 plots, respectively, because their staggers scores were ≥4. The cumulative probability of lambs having scores ≥4 did not differ between animals grazing the two pasture types (p=0.41).

CONCLUSIONS AND CLINICAL RELEVANCE: There was no evidence for ergovaline increasing the severity of ryegrass staggers induced by lolitrem B. In situations where the severity of ryegrass staggers appears to be greater than that predicted on the basis of concentrations of lolitrem B, the presence of other tremorgenic alkaloids should be investigated.     

Improved micropropagation protocol and enhancement in biomass and chlorophyll content in Stevia rebaudiana (Bert.) Bertoni by using high copper levels in the culture medium

Incorporation of a range of higher concentrations of CuSO₄·5H₂O in MS medium [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassay with 240 tobacco tissue cultures. Physiol. Plant 15, 473–497] significantly enhanced direct shoot bud induction and proliferation from cultured leaf and nodal explants taken from mature plants of Stevia rebaudiana. Shoot bud induction medium was supplemented with BAP (2.2 μM) and IAA (2.8 μM). When the concentration of CuSO₄·5H₂O in the induction medium was raised to 0.5 μM (five times the MS level, i.e. 0.1 μM) there was significant increase in percentage response along with increase in shoot bud number per explant. The shoots were healthy, well developed with dark green broader leaves. There was remarkable increase in total biomass and chlorophyll content at increased (0.5 μM) copper level in the medium. During proliferation stage also presence of high copper levels in the medium favoured increase in shoot bud number per explant.     

Methane emission from two Indian soils planted with different rice cultivars

In a greenhouse study, methane emissions were measured from two diverse Indian rice-growing soils planted to five rice cultivars under similar water regimes, fertilizer applications and environmental conditions. Significant variations were observed in methane emitted from soils growing different cultivars. Total methane emission varied between 8.04 and 20.92gm^–2 from IARI soil (Inceptisol) and between 1.47 and 10.91gm^–2 from Raipur soil (Vertisol) planted to rice. In all the cultivars, emissions from IARI soil were higher than from Raipur soil. The first methane flux peak was noticed during the reproductive phase and the second peak coincided with the grain-ripening stage of the rice cultivars. Received: July 7, 1996     

The Prospects of Tissue Culture and Genetic Engineering for Strawberry Improvement

Abstract

Strawberry shoot meristem cultures have been used for producing virus-free strawberry plants. Plantlet regneration from leaf mesophyll protoplasts, and Agrobacterium tumefaciens mediated genetic transformation, mark the beginning of strawberry improvement with biotechnologies. The use of biotechnological tools has a greater potential in producing strawberry plants with traits such as early maturation, frost and disease resistance, mite and nematode resistance, high yield, better quality, suitability for mechanical harvesting, shipping ability, ellgaic acid content, aroma compounds and cost-effective micropropagation. Gene identification and mapping with RFLPs or RAPDs would be desirable. Transgenic strawberries should be tested for their quality before releasing to the consumers.