Haematological assessment of the effects of the anaesthetic MS 222 in natural and neutralized form in three freshwater fish species: interspecies differences

Interspecies haematological differences to MS 222 and neutralized MS 222 anaesthesia were investigated in Sarotherodon mossambicus, Cyprinus carpio and Salmo gairdneri acclimated under identical laboratory conditions. Anaesthesia with MS 222 resulted in a 'chemical stress' in all fish, as was evident from changes in the haematological profiles of the animals. Such species specific variations in the haematology persisted throughout the whole experiment protocol which employed different concentrations of the anaesthetic. The use of neutralized MS 222, whereby aquarium water quality remained unchanged, improved the haematological profile. Possible reasons for the interspecies differences observed are discussed.     

Methods of minimal residual disease (MRD) detection in childhood haematological malignancies

The appropriate management of haematological disorders must rely on a precise and long-term monitoring of the patient's response to chemotherapy and radiotherapy. Clinical data are not sufficient and that is why in the last decade it became the most important to improve the knowledge of haematological diseases on the basis of molecular techniques and molecular markers. The presence of residual malignant cells among normal cells is termed minimal residual disease (MRD). Nowadays a great progress has been made in the treatment of malignant diseases and in the development of reliable molecular techniques, which are characterised by high sensitivity (10-3- 10-6) and ability to distinguish between normal and malignant cells at diagnosis and during follow-up. Especially, MRD data based on quantitative analysis (RQ-PCR, RT-RQ-PCR) appear to be crucial for appropriate evaluation of treatment response in many haematological malignancies. Implementation of standardized approaches for MRD assessment into routine molecular diagnostics available in all oncohaematological centres should be regarded nowadays a crucial point in further MRD study development.     

The effect of bacterial infection on erythrocyte fragility and sedimentation rate of rainbow trout, Salmo gairdneri Richardson

Osmotic fragilities and erythrocyte sedimentation rates (ESR) were measured in healthy, diseased, treated and recovered trout, Salmo gairdneri . Bacterial infection with Aeromonas and Streptococcus resulted in more fragile erythrocytes and raised ESR values. Treatment with antibacterial agents improved the values of both parameters. Possible reasons for such changes are discussed. Caution in the use of apparently healthy fish for the unqualified interpretation of haematological results is expressed.     

Clinico-pathological studies on naturally-occurring bovine fascioliasis in the Sudan.

An investigation was made of the pathological, haematological and biochemical aspects of naturally-occurring bovine fascioliasis in the Sudan. 228 animals infected with Fasciola gigantica and 25 non-infected controls were used in the study. The infected cattle revealed emaciation, typical liver pathology, and, occasionally, lesions in the lung and the pancreas. Analysis of their sera also showed reduced albumin values, increased globulin concentrations and decrease albumin/globulin ratio, in addition to increased arginase activity. The serum iron concentration, on the other hand, was decreased, while the total iron binding capacity increased and the resultant iron saturation values reduced. Haematological findings in the infected animals included reduced erythrocyte counts, decreased haematocrit values, increased mean corpuscular volumes, eosinophilia and decreased neutrophil, lymphocyte and monocyte counts. Other parameters were similar in infected and control cattle.     

Effects of ketamine anaesthesia, stress and repeated bleeding on the haematology of vervet monkeys

Haematology values are presented for the vervet monkey (Cercopithecus aethiops), and the relative effects of high dose ketamine anaesthesia, stress of capture and repeated bleedings assessed. Anaesthesia resulted in decreased WBC and RBC values, attributed to depression of cardiovascular function. These effects were the reverse of those of alarm and strenuous exercise (leukocytosis and polycythaemia) during capture. Stress resulted in relatively high white and low red blood cell counts. Opposing effects of stress and anaesthesia led to comparable haematological values for trained, non-anaesthetized vervets and stressed, anaesthetized vervets. Effects of repeated bleedings were opposite in anaesthetized and non-anaesthetized animals. These effects, however, along with those of ketamine anaesthesia and stress, were relatively insignificant compared with the wide variation in haematological values found among individuals. The biological importance of these effects thus appeared to be slight. The concept of 'normal values' is discussed.     

Evaluation of lung diffusing capacity by physiological and morphometric techniques

Determinations of pulmonary diffusing capacity for CO (DLCO) by physiological and morphometric techniques have resulted in substantially different values for both DLCO and its major components. To evaluate the differences in these methods of measurement of DLCO, measurements were made under controlled conditions on isolated perfused dog lungs. Multiple gas-rebreathing techniques were used to measure DLCO, the membrane component of the diffusing capacity for CO (DmCO), and pulmonary capillary blood volume (Vc) in both anesthetized dogs and after isolation and perfusion of their lungs. The isolated perfused lungs were than perfusion fixed for morphometric analysis of the components of DLCO. The values obtained morphometrically for Vc were similar to those measured by physiological techniques. Perfusion fixation did not substantially alter the morphometric estimate of DmCO when compared with previous values obtained on inflation fixed lungs. However, the morphometric estimate of DmCO was over 10 times higher than that estimated physiologically. Analysis of the potential errors in the techniques suggests that the correct value for DmCO is substantially higher than that commonly estimated by use of physiological techniques and that the explanation for the difference is due to a number of factors that can influence the binding of CO to hemoglobin under in vivo conditions. The net effect of these factors can be represented by an unknown in each component of the Roughton-Forster relationship so that 1/DL = 1/(U1.Dm) + 1/(U2.theta Vc), where theta is the binding rate for CO to hemoglobin. Because the magnitudes of the unknown terms (U1 and U2) in the Roughton-Forster relationship are likely to be large, this relationship cannot be reliably used to determine Dm and Vc.     

Studies on the manufacture of hematological and cytological specimens using the centrifugation technic]

Centrifugal preparation (Cp) represents a method of creating haematological parameters which, in addition to a qualitative improvement of representing single cells, is suitable to produce reproducible haematological smear preparations. In qualitative and quantitative respect centrifugal preparations will reflect the intravasal conditions better than the common blood smear technique. Further advantages are the concentration of cellular elements and in connection with it the opportunity of finding cells rarely present. With a small amount of work centrifugal preparations enable counting values to be established as well as cytological and cytochemical findings from the venous blood to be obtained. As a result of the test it turned out that the common glass slides cannot be replaced by plastic foils ones without certain disadvantages appearing. First investigations of attempting to represent cells in culture medium as well as analyzing liquors and processing other biological liquids with the help of the centrifugal preparation technique brought unsatisfactory results. Centrifugal preparation provides the possibility of performing pre-programmed haematological examinations beginning with blood collection and ending up in the evaluation by a computer.     

Mathematical modeling of the dynamics of the intensity of the mutation process. I. Formulation of the task and the general model

Methodology is devised to construct dynamic models of the mutation process intensity under quasi-stationary environmental conditions and limited experimental information. Laws of distribution of unknown parameters of dynamic models and prognostic values of inherited pathology frequencies were determined for the cases of normal distribution with zero mathematical expectancy and constant dispersion of casual environmental influence. Given is the algorithm of dynamic model construction on frequencies of inherited pathology, using experimental data and on the basis of determination of permissible intervals for their prognostic values.     

Comparative genomic hybridisation for the analysis of chromosomal imbalances in solid tumours and haematological malignancies

 Comparative genomic hybridisation (CGH) is based on a two-colour, competitive fluorescence in situ hybridisation of differentially labelled tumour and reference DNA to normal metaphase chromosomes. This new technology has made a great impact in molecular tumour pathology due to its possible application to archival specimens and the ability to create copy number karyotypes throughout the whole genome from very small amounts of DNA. If chromosomal imbalances can be correlated with a etiological and clinical features of tumours, CGH could be able to provide new prognostic and diagnostic criteria. CGH findings further provide starting points for the molecular genetic characterisation of altered chromosomal regions harbouring yet unidentified genes involved in tumorigenesis and tumour progression. An overview of the results of published CGH studies on solid tumours and haematological malignancies is presented. Methodological limitations of the CGH technology are reported, as well as future developments which will improve its use in routine analysis. Accepted: 29 July 1997     

Blood biochemical indicators in young and adult Cebus apella of both sexes

BACKGROUND: A frequent drawback in physiology of non-human primates is that normal values for a variety of indicators (haematological, biochemical and others) are scant. METHODS: We report here the blood values in a series of 92 healthy Cebus apella (divided by sex, age and pregnancy status). Health check-ups indicated that animals were healthy for the month prior to and after the sampling. Dietary intake was estimated on the basis of two semi-balance studies. RESULTS: Values of haematological indicators, serum LDH activity, micronutrient indicators (serum copper, iron and serum ceruloplasmin concentrations, Zn-Cu-superoxide dismutase activity in erythrocytes) agreed with previous results and provide some values that were not available. Activities of liver enzymes were lower than some previously reported. CONCLUSIONS: These results provide valuable information that help understanding the physiology of C. apella.     

Tissue microarrays: fast-tracking protein expression at the cellular level

Tissue microarrays maximize returns in cellular pathology whilst minimizing the use of cells and tissues. They are made by arraying cores of tissue taken from multiple donor blocks into a single recipient block. Accordingly, the histology and pathology of several hundred tissues can be represented in one tissue microarray that, when stained by immunohistochemistry, provides comprehensive topographic information on protein expression. Used with complimentary techniques, such as complementary DNA microarray analysis, tissue microarrays are providing valuable data for the identification of new markers of disease and assisting in the discovery of therapeutic targets. They are also leading a revolution in cellular pathology as high-throughput technology is introduced to maximize the information provided.     

Tissue microarrays: fast-tracking protein expression at the cellular level

Tissue microarrays maximize returns in cellular pathology whilst minimizing the use of cells and tissues. They are made by arraying cores of tissue taken from multiple donor blocks into a single recipient block. Accordingly, the histology and pathology of several hundred tissues can be represented in one tissue microarray that, when stained by immunohistochemistry, provides comprehensive topographic information on protein expression. Used with complimentary techniques, such as complementary DNA microarray analysis, tissue microarrays are providing valuable data for the identification of new markers of disease and assisting in the discovery of therapeutic targets. They are also leading a revolution in cellular pathology as high-throughput technology is introduced to maximize the information provided.     

A technique for repeated sampling of the blood of individual resting fish.

A dorsal cannulation technique is described. It has been employed for repeated blood sampling in unanaesthetized rainbow trout (Salmo gairdneri) kept singly in special receptacles described in the paper. The level of the studied haematological parameters (Haematocrit, Hb, glucose, lactate, K+, Na+, Ca2+) differed between fish kept in receptacles for 1 week and free-swimming fish, most probably owing to differences in the motility of the fish. The receptacle seems to minimize visual and handling disturbances, and permits both the standardization of experimental conditions and quick and easy sampling via the dorsal aorta cannula. The general variation in the blood parameter values was very small compared with the previously reported variation in such values for rainbow trout.     

Seasonal changes in the haematological indices in peripheral blood of chinchilla (Chinchilla laniger L.)

Changes in nine haematological indices in adult female and male chinchilla were investigated over a period of 18 months. All haematological values were in the same range both for females and males, although seasonal fluctuations were more evident in the females. High levels of the red blood cells count (RBC), haemoglobin concentration (Hb), mean cell haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC) during the winter months and low levels of these parameters during the summer months were found. The white blood cells count (WBC) and lymphocyte number (L) also decreased in summer, their values increasing during autumn and spring. The haematocrit values (Ht) and mean cell volumes (MCV) are high during the summer and low during the winter.     

Comparative haematology of ten species of marine fish from Visakhapatnam Coast

A comparative study has been made of haematological parameters (erythrocyte numbers, leucocyte numbers, haematocrit and haemoglobin), erythrocyte indices (MCV, MCH and MCHC) and cytometric measurements (cell and nucleus) of 10 species of marine fish from the Visakhapatnam Coast, of which six are from harbour waters, three from rock pools and one from inshore waters. It is found that active fish have high values of erythrocyte numbers, haematocrit, haemoglobin and nucleo/cytoplasmic ratio and low values of leucocyte numbers and erythrocyte cytosome measurements. The most active of all is the inshore species Rastrelliger kanagurta depicting the haematological requirements of an active fish with high oxygen demand to meet the requirements of high metabolic rate. The other active fish such as Megalops cyprinoides and Mugil cephalus are intermediate, with the rock pool fish and Caranx carangus with low values of nucleo/cytoplasmic ratio at the other extreme.     

The haematology of Rohu, Labeo rohita

Haematocrit values, haemoglobin concentration, erythrocyte counts, leucocyte counts, erythrocyte sedimentation rate and clotting time of Labeo rohita were determined. The variations in haematological parameters in relation to the size and weight of the fish were also examined. Males had higher haematological values than the females. Seasonal and variations due to maturation of gonads were also apparent.     

Functional Analysis: Evaluation of Response Intensities - Tailoring ANOVA for Lists of Expression Subsets

Background  

Microarray data is frequently used to characterize the expression profile of a whole genome and to compare the characteristics of that genome under several conditions. Geneset analysis methods have been described previously to analyze the expression values of several genes related by known biological criteria (metabolic pathway, pathology signature, co-regulation by a common factor, etc.) at the same time and the cost of these methods allows for the use of more values to help discover the underlying biological mechanisms.     

Haematological profile in healthy urban population (8 to 70 years of age)

Haematological profile for 17 constituents of blood were determined in 998 healthy school children (8-19 years old) and 2246 healthy adult persons (20-70 years old) residing permanently or at least 5 years in a defined geographic region of Zagreb, Croatia. Physiological variations corresponding to age and sex were studied as the most important factors affecting biological variation in haematological constituents of blood. In our study erythrocytes, haemoglobin and haematocrit values were not sex dependent until the age of 13 after which the values were higher in men that in women. Sedimentation rate showed sex and age related differences in the adult age with higher values in women especially after 50 years. Total leukocyte count declined with age and in adults the values were slightly lower in women. Segmented neutrophil granulocytes showed the upward trend with age whereas the lymphocyte and monocyte counts declined. Women showed slightly higher platelet count in the adult age. Based on biological variation, we have estimated the reference intervals for 17 haematological constituents of blood in order to provide medically reliable evaluation of haematological laboratory results.