Flow cytometric patterns in blood from dogs with non-neoplastic and neoplastic hematologic diseases using double labeling for CD18 and CD45

BACKGROUND: In dogs, flow cytometry is used in the phenotyping of immunologic cells and in the diagnosis of hemic neoplasia. However, the paucity of specific antibodies for myeloid cells and B lymphocytes and of labeled antibodies for multicolor techniques limits the ability to detect all leukocyte subpopulations. This is especially true for neoplastic and precursor cells. CD18 and CD45 are expressed on all leukocytes and are involved in cell activation, and together could be useful in helping determine cell lineage. OBJECTIVES: The purpose of this study was to double label canine blood for CD18 and CD45 and to use the differential expression of antigens to identify leukocyte populations in dogs with non-neoplastic and neoplastic hematologic diseases. METHODS: A template was developed using blood samples from 10 clinically healthy dogs and a back-gating technique. Differential leukocyte counts obtained with the template were compared with those obtained by manual and automated methods on blood samples from 17 additional healthy dogs. Blood samples obtained from 9 dogs with non-neoplastic (reactive) hematologic diseases and 27 dogs with hemic neoplasia were double stained for CD18 and CD45 using mouse anticanine CD18 monoclonal antibody (mAb) plus phycoerythrin-conjugated rat anticanine CD45 mAb and fluorescein isothiocyanate-conjugated rabbit antimouse IgG. Hemic neoplasms were diagnosed by cell morphology, and immunophenotypic and cytochemical markers. RESULTS: With the double label, neutrophils, eosinophils, monocytes, and T- and B-lymphocytes were identified. In reactive disorders, a population of activated neutrophils with high CD45 and CD18 expression was detected. In hemic neoplasia, cell lineage was easily determined, even in acute leukemia. CONCLUSIONS: Double labeling for CD18/CD45 may be useful as a screening method to evaluate hematologic diseases and help determine cell lineage, and to aid in the selection of a panel of antibodies that would be useful for further analysis.     

Multi-electrode 3D resistivity imaging of alfalfa root zone

Information on the amount and spatial distribution of plant roots is increasingly needed for understanding and managing crop behaviour. Soil electrical resistivity (ρ) tomography has been proposed as a non-destructive method for root biomass quantification and mapping in trees but evidence is needed on the applicability of the technique at low root density and in herbaceous plants.We produced high-resolution 3D DC soil resistivity tomograms in containers with bare soil (B), and alfalfa (Medicago sativa L.) (A1) on a silt loam soil, and alfalfa on a loam (A2). Root biomass (RMD), root length density (RLD), soil electrical conductivity (EC) and water content (θ) were measured destructively.The pattern of soil resistivity matched the spatial distribution of θ in bare soil and of RMD in rooted soil. Univariate linear relations were found between ρ and θ in bare soil and between ρ, RLD and RMD in rooted soil. Across all data RMD and soil texture (P < 0.01) explained a high proportion of variability in soil resistivity.This allows to conclude that soil resistivity is quantitatively related to root biomass in herbaceous plants even at low root density (biomass < 0.001 Mg m⁻³), providing a basis for the development of resistivity-founded methods for the non-destructive spatial detection of root mass in situ, but the response in ρ is of the same order of magnitude as the effects of grain size and water content. Therefore in field studies reciprocal masking of low-density roots and other soil features is possible, and the effect of variation in other soil properties should be explicitly addressed.     

Chalinulasterol, a chlorinated steroid disulfate from the Caribbean sponge Chalinula molitba. Evaluation of its role as PXR receptor modulator

Chalinulasterol (1) a new chlorinated sterol disulfate was isolated from the Caribbean sponge Chalinula molitba. Its structure was elucidated using mass spectrometry and NMR experiments. The possible role of chalinulasterol as modulator of the PXR nuclear receptor was investigated but, in spite of the close structural relationship with the PXR agonist solomonsterol A (2), it showed no activity. The structural requirements for the PXR nuclear receptor activity were discussed.     

Effects of Reduced Carbonate Saturation State on Early Development in the Common Edible Sea Urchin Lytechinus variegatus: Implications for Land-Based Aquaculture

Land-based aquaculture facilities often utilize additional bicarbonate sources such as commercial sea salts that are designed to boost alkalinity in order to buffer seawater against reductions in pH. Despite these preventative measures, many facilities are likely to face occasional reductions in pH and corresponding reductions in carbonate saturation states due to the accumulation of metabolic waste products. We investigated the impact of reduced carbonate saturation states (Ω_Ca, Ω_Ar) on embryonic developmental rates, larval developmental rates, and echinoplutei skeletal morphometrics in the common edible sea urchin Lytechinus variegatus under high alkalinity conditions. Commercial artificial seawater was bubbled with a mixture of air and CO₂ gas to reduce the carbonate saturation state. Rates of embryonic and larval development were significantly delayed in both the low and extreme low carbonate saturation state groups relative to the control at a given time. Although symmetry of overall skeletal body lengths was not affected, allometric relationships were significantly different between treatment groups. Larvae reared under ambient conditions had significantly greater postoral arm and overall body lengths relative to body lengths than larvae grown under extreme low carbonate saturation state conditions, indicating that extreme changes in the carbonate system affected not only developmental rates but also larval skeletal shape. Reduced rates of embryonic development and delayed and altered larval skeletal growth are likely to negatively impact larval culturing of L. variegatus in land-based, intensive culture situations where calcite and aragonite saturation states are lowered by the accumulation of metabolic waste products.     

Molecular mechanism regulating axoneme activation in marine fish: a review

River Gomti, a tributary of river Ganga in northern India, is being polluted due to indiscriminate disposal of domestic sewage and industrial wastes that contain genotoxic chemicals. The study was conducted to evaluate the genotoxic potential of polluted water of river Gomti in two fish species, namely Channa punctatus and Mystus vittatus. The fishes were exposed in situ in nylon cages to the polluted water of river Gomti fixed near a distillery outlet located in Lucknow. The induction of DNA damage and micronuclei were determined in blood erythrocytes using comet assay and micronucleus test, respectively. The induction in micronuclei frequencies and DNA damage were found to be significantly elevated (p?<?0.01) in exposed specimens after 3?days post-exposure as compared to the control, i.e. from laboratory-acclimatized fish specimens. The comparison of DNA damage between the two species indicated that C. punctatus is more sensitive to aquatic pollutants. Thus, this fish could be used as a bio-indicator of genotoxicity for bio-monitoring of water bodies. The results further revealed that the river Gomti is being contaminated with potential genotoxic and mutagenic chemicals produced from industrial and domestic activities; therefore, immediate measures are needed to reduce the inflow of pollutants in the river.     

Vibrio spp. Control at Brine Shrimp,Artemia, Hatching and Enrichment

Experiments were conducted to evaluate different prophylactic methods to control the bacterial load in brine shrimp, Artemia, hatching. The first experiment evaluated three treatments to control Vibrio spp. during the Artemia hatching: microalgae (Chaetoceros calcitrans), probiotic (Bacillus spp.), and antibiotic (Florfenicol). In the second experiment, Artemia metanauplius were enriched in distinct treatments with C. calcitrans, probiotic, and emulsion rich in docosahexaenoic and eicosapentaenoic fatty acids. Enriched Artemia metanauplius and nauplii (control) were offered to white shrimp, Litopenaeus vannamei, postlarvae (PL₇–PL₁₉). Presumptive Vibrio were quantified in Artemia, PL, and rearing water. Microalgae and probiotic were effective to control Vibrio spp. in Artemia nauplii. The enrichment process increased the Artemia bacterial load but did not affect Vibrio load in L. vannamei.     

Antimycotic effectiveness against dermatophytes: comparison of two in vitro tests

Seven antimycotic drugs (econazole, enilconazole, fluconazole, griseofulvin, itraconazole, ketoconazole, and miconazole) were tested against 36 dermatophyte strains (19 M. canis, 7 T. mentagrophytes, 5 M. gypseum, 2 M. cookei, 1 T. rubrum, 1 T. ajelloi, and 1 T. terrestre) isolated from animals, humans, and the environment. Two in vitro methods were compared: a micro-dilution test based on the CLSI M38-A method, and a disk-diffusion test. Fluconazole was not effective in vitro against the dermatophytes. Econazole and enilconazole were the most effective. Thirteen strains were griseofulvin-resistant. The correlation between the two methods was statistically significant for enilconazole, griseofulvin, itraconazole, and miconazole.     

The essential oil of Eucalyptus tereticornis, and its constituents α- and β-pinene,potentiate acetylcholine-induced contractions in isolated rat trachea

The effects of the essential oil of Eucalyptus tereticornis (EOET), especially the effects of its constituents α- and β-pinene, were studied on rat trachea in vitro. In tracheal rings, EOET, α- or β-pinene potentiated the contractions induced by acetylcholine (ACh). Contractions induced by K⁺ (60 mM) were also potentiated by α- and β-pinene, but were reduced by EOET. Our findings show that EOET has myorelaxant effects on rat airways, but potentiates ACh-induced contractions. Monoterpenes α- and β-pinene are involved in its potentiating actions, but are not responsible for its myorelaxant effects. A putative inhibition of the acetylcholinesterase enzyme is involved.     

First report of the use of meglumine antimoniate for treatment of canine leishmaniasis in a pregnant dog

Canine leishmaniasis during pregnancy is rarely reported, even in countries where the infection in dogs is endemic. The authors report a case of a 4 yr old bitch with leishmaniasis treated with meglumine antimoniate during pregnancy. The pregnancy and delivery were normal and the bitch presented improvement of the infection during treatment. Three puppies died within 2 days of birth and tested negative via real-time PCR for L. infantum. The two surviving puppies were followed clinically, serologically, and by real-time PCR until 1 yr of age with no evidence of congenital leishmaniasis. L. infantum DNA was detected with real-time PCR analysis of uterine tissue from the bitch at the time of ovariohysterectomy. PCR analysis was performed after an ovariohysterectomy of the bitch that was performed two months after parturition. Meglumine antimoniate use in the pregnant bitch may have prevented vertical transmission of leishmaniasis.