Haematological and haematopoietic studies in pyridoxine deficient fish, Channa punctatus Bloch

Pyridoxine deficiency was induced in a population of Channa punctatus. The deficiency effects were increased by adding 4-deoxypyridoxine as an antagonist. Haematological and haematopoietic changes were studied. The significant changes in peripheral blood appeared after 150 days in the antagonist treated group and 180 days in the group on the deficient diet. After 240 days when morphological changes and an increased mortality also appeared, the changes in RBC and related values (haemoglobin, haematocrit, MCV, MCH & MCHC) were highly significant and indicated hypochromic microcytic anaemia; this was accompanied by leukopenia. The haematopoietic studies revealed significant increase in the earlier stages of erythrocytic development (basophilic erythroblasts and polychromatophilic eryth-roblasts) with a simultaneous decrease in the later stages (young and mature reticulocytes). A series of pathological disorders in the various developing cells were recorded in the imprints of haematopoietic tissues. The pyridoxine specificity of these changes were confirmed by remedial treatment for a month.     

Insights into the biology of cord blood stem/progenitor cells

OBJECTIVES: To review information on cord blood banking and transplantation with respect to the author's studies, and in context of this field of investigation. RESULTS: Cord blood transplantation has been successfully used to treat a number of malignant and non-malignant disorders. However, this technique is still associated with limited numbers of cells for transplantation, and with delayed engraftment of neutrophils and platelets. The field of cord blood transplantation will benefit from enhanced and mechanistically based information on haematopoietic stem cell function and potential means to enhance its effectiveness are reviewed. This includes notions concerning possibility of retrieving more cells from the placenta and cord blood, to expand haematopoietic stem cells ex vivo and to increase efficiency of homing and engraftment of these cells. Also discussed are cryopreservation and long-term storage of cord blood haematopoietic and progenitor cells, and new laboratory findings and animal studies for non-haematopoietic uses of cord blood.     

Nature of the air-breathing organs of the Indian fishes Channa, Amphipnous, Clarias and Saccobranchus as shown by electron microscopy

The structure of the air-breathing organs of the Indian fishes Channa punctatus, Channa Striatus, Amphipnous cuchia, Clarias batrachus and Saccobranchus fossilis has been investigated using electron microscopy. In all species the barrier separating the air from the blood consists of three main layers (epithelium, basal lamina and endothelium). The total thickness ranging from 0.78 μm in C. punctatus 1.6 μm in S. fossilis.
In Clarias and Saccobranchus the presence of pillar cells characteristic of gill secondary lamellae confirms evidence for the origin of these organs by modification of a typical gill structure.
In Amphipnous and two species of Channa , however, the evidence suggests that the accessory organs represent modified gills. The presence of valve-like structures between the afferent and efferent blood spaces of the vascular papillae gave the appearance of pillar cells under the light microscope.
The structure of these organs is correlated with physiological studies on the degree of their importance in the life of the animal and the degree of gill development     

Haematopoietic stem cell niche in Drosophila

Development and homeostasis of the haematopoietic system is dependent upon stem cells that have the unique ability to both self-renew and to differentiate in all cell lineages of the blood. The crucial decision between haematopoietic stem cell (HSC) self-renewal and differentiation must be tightly controlled. Ultimately, this choice is regulated by the integration of intrinsic signals together with extrinsic cues provided by an exclusive microenvironment, the so-called haematopoietic niche. Although the haematopoietic system of vertebrates has been studied extensively for many decades, the specification of the HSC niche and its signals involved are poorly understood. Much of our current knowledge of how niches regulate long-term maintenance of stem cells is derived from studies on Drosophila germ cells. Now, two recently published studies by Mandal et al.1 and Krezmien et al.2 describe the Drosophila haematopoietic niche and signal transduction pathways that are involved in the maintenance of haematopoietic precursors. Both reports emphasize several features that are important for controlling stem cell behavior and show parallels to both the vertebrate haematopoietic niche as well as the Drosophila germline stem cell niches in ovary and testis. The findings of both papers shed new light on the specific interactions between haematopoietic progenitors and their microenvironment.     

Regenerative capacity of the spleen in a splenectomized fish, Channa punctatus Bloch, with related investigations into changes in peripheral blood and haematopoiesis

The present investigation has provided evidence by a simultaneous study of the regenerating spleen, peripheral blood and the kidney of the haematopoietic tissue to establish the role of the spleen in the development of circulating blood cells in Channa ( Ophiocephalus ) punctatus Bloch. Regeneration of the spleen after eight to nine weeks of splenectomy was shown to be complete. Weekly observations a fortnight after splenectomy are recorded with reference to spleen morphology and weight, erythrocyte and differential leucocyte counts, haemoglobin content, haematocrit value and related red cell indices (MCV, MCH, MCHC). The results show that splenectomy leads to a macrocytic hypochromic anaemia and leucopenia, resulting in 25% reduction in haemoglobin content and a decrease in leucocyte numbers in five weeks following splenectomy. Thrombocytes and neutrophils show a relative ( P < 0·01) increase. The leucopenia is caused by large and small lymphocytes, eosinophils and basophils whose number was reduced from 50% to 67%.     

Effect of streptozotocin on the blood glucose level and histology of the principal islets of Channa punctatus (Bloch).

The effect of streptozotocin, an antibiotic and diabetogenic drug, has been studied on the blood glucose level and islet histology of a freshwater fish, Channa punctatus. The drug elicits a triphasic response in the blood glucose level, comprising an initial hyperglycemia followed by a transient fall, and restitution of normal values 3...4 days after the treatment. Significant degenerative changes occur in the islet beta cells. Severity of the beta cell damage is dose dependent and the drug has been found to be beta-cytotoxic to a considerable extent. Unlike mammals, Channa punctatus does not become diabetic following the streptozotocin administration, at doses varying 200-400 mg/kg b.wqnd over a period of 96 hours post-injection.     

Molecular characterization of eight Indian Snakehead species (Pisces: Perciformes Channidae) using RAPD markers

Murrels (Perciformes; Channidei; Channidae) are unique group of freshwater air breathing fishes having a confined distribution to African and Asian continents. The phylogenetic relationship among eight Channid species viz. Channa aurantimaculata, Channa bleheri, Channa diplogramma, Channa gachua, Channa marulius, Channa punctatus, Channa stewartii and Channa striatus were investigated using RAPD markers. Eight random oligodecamers viz. OPAC03, OPAC05, OPAC07, OPAC09, OPAC19, OPA10, OPA11 and OPA16 were used to generate the RAPD profile. Estimates of Nei’s (Genetics, 89:583–590, 1978) unbiased genetic distance (D) demonstrated sufficient genetic divergence to discriminate the samples of different species and the values ranged from 0.3292 to 0.800 The present RAPD analyses strongly substantiate the view of earlier morphological and osteological studies of Channid species, the closer association among species in “gachua” and “marulius” groups.     

Inflammatory modulation of HSCs: viewing the HSC as a foundation for the immune response

Cells of the innate and adaptive immune systems are the progeny of a variety of haematopoietic precursors, the most primitive of which is the haematopoietic stem cell. Haematopoietic stem cells have been thought of generally as dormant cells that are only called upon to divide under extreme conditions, such as bone marrow ablation through radiation or chemotherapy. However, recent studies suggest that haematopoietic stem cells respond directly and immediately to infections and inflammatory signals. In this Review, we summarize the current literature regarding the effects of infection on haematopoietic stem cell function and how these effects may have a pivotal role in directing the immune response from the bone marrow.     

Haematological and haematopoietic response to acid stress in an air-breathing freshwater fish, Channa punctatus Bloch

Response of Channa punctatus to acidic water was studied by exposing fishes to pH 3.5, 4.5, 5.5 and 6.5 for 6 weeks. Growth and mortality data indicated increasing stress as the acid level in the ambient water increased. While no mortality was recorded at pH 6.5, a distinct loss of weight compared to continuous gain in body weight in control fish indicated stress. As the pH level decreased, the rate of loss in body weight increased accompanied by mortality which rose to as high as 60% within 3 weeks in fishes exposed to water at pH 3.5.
Haematological investigations confirmed the general stress indicated by growth and mortality data. Thus. RBC and related values indicated overall polycythemia. However, eosinophils, basophils. and large and small lymphocytes showed a distinct fall in number as compared to the control.
Correlated haematopoietic studies revealed that both the initial and penultimate stages in RBC and neutrophil development recorded an increase parallel to that observed in peripheral blood, but intermediate stages, probably because they were unable to keep pace with the fast turnover, showed a relative decrease.
Biochemical investigations showed an increase not only in blood glucose level but also in liver glycogen content. However, there was a significant decrease in muscle glycogen reserves.
The significance of these changes is discussed.     

WNT signalling and haematopoiesis: a WNT-WNT situation

The evolutionarily conserved WNT-signalling pathway has pivotal roles during the development of many organ systems, and dysregulated WNT signalling is a key factor in the initiation of various tumours. Recent studies have implicated a role for WNT signal transduction at several stages of lymphocyte development and in the self-renewal of haematopoietic stem cells. Here, we outline new insights into the WNT-signalling pathway, review its role in the self-renewal of haematopoietic stem cells and in the development of T and B cells, and discuss controversies and future developments with regard to WNT signalling in the thymus.     

Evaluation of neural plasticity in adult stem cells

The role of stem cells has long been known in reproductive organs and various tissues including the haematopoietic system and skin. During the last decade, stem cells have also been identified in other organs, including the nervous system, both during development and in post-natal life. More recently, evidence has been presented that stem cells thought to be responsible for the generation of mature differentiated cells of one organ, such as haematopoietic stem cells, may have the ability to also differentiate across lineages and contribute to tissues other than haematopoietic cells, including neuronal tissue, suggesting that easily accessible stem cells sources may one day be useful in the therapy of ischaemic (stroke) and also degenerative diseases of the nervous system. Here, we will evaluate the validity of such claims based on a number of criteria we believe need to be fulfilled to definitively conclude that certain stem cells can give rise to functional neural cells that might be suitable for therapy of neural disorders.     

Genetic relatedness among fish species of Genus Channa using mitochondrial DNA genes

The family Channidae is represented by 26 species, out of which 23 species are found in Asia. However, the taxonomy and phylogeny of the Channid fishes found in India are poorly understood. In the present study, eight species of Channa (Channa striata, Channa punctatus, Channa marulius, Channa gachua, Channa stewartii, Channa aurantimaculata, Channa barca and Channa bleheri) were investigated using partial sequences of 16S rRNA and Cytochrome c Oxidase subunit I (COI) of mitochondrial genes to differentiate among the eight species and study their relationships. The sequence analysis of the genes revealed two distinct groups, which are genetically distant from each other and exhibit identical phylogenetic resolution. The partial sequences of both the genes provided sufficient phylogenetic information to distinguish all the eight species of Channa.     

Genetic diversity and morphological stasis in the Ceylon Snakehead,Channa orientalis (Teleostei: Channidae)

Ichthyological Research - As the type species of the genus Channa, the identity of the pelvic-finless snakehead Channa orientalis Bloch is important to channid systematics. Although this name has...     

The role of phosphotyrosine phosphatases in haematopoietic cell signal transduction

Phosphotyrosine phosphatases (PTPases) are the enzymes which remove phosphate groups from protein tyrosine residues. An enormous number of phosphatases have been cloned and sequenced during the past decade, many of which are expressed in haematopoietic cells. This review focuses on the biochemistry and cell biology of three phosphatases, the transmembrane CD45 and the cytosolic SH2-domain-containing PTPases SHP-1 and SHP-2, to illustrate the diverse ways in which PTPases regulate receptor signal transduction. The involvement of these and other PTPases has been demonstrated in haematopoietic cell development, apoptosis, activation and non-responsiveness. A common theme in the actions of many haematopoietic cell PTPases is the way in which they modulate the thresholds for receptor signalling, thereby regulating critical events in the positive and negative selection of lymphocytes. There is growing interest in haematopoietic PTPases and their associated regulatory proteins as targets for pharmaceutical intervention and in the involvement of these enzymes in human disease.     

Wiskott-Aldrich syndrome protein, WASP

Wiskott-Aldrich Syndrome protein (WASP) is the product of the gene mutated in children with Wiskott-Aldrich Syndrome (WAS). It is a predominantly cytoplasmic protein, expressed only in haematopoietic cells. It binds in vivo to the adaptor proteins Nck and Grb2, to the cytoplasmic protein-tyrosine kinase Fyn and to the small Rho-like GTPase Cdc42, which is required for formation of filopodia in fibroblasts and macrophages. WASP also interacts, directly or indirectly, with the actin cytoskeleton. Together with studies of a closely related, ubiquitously expressed protein named N-WASP, these findings suggest that WASP is a component of signalling pathways that control reorganisation of the actin cytoskeleton in haematopoietic cells in response to external stimuli. In support of this idea, haematopoietic cells from WAS patients show defects in cytoskeletal organisation that compromise their ability to polarise and to migrate in response to physiological stimuli. These defects could account for many of the clinical features of WAS. WAS is now a candidate for gene therapy based on the delivery of a wild-type WASP gene to autologous haematopoietic stem cells. In addition, recent studies of cell defects in WAS patients suggest that it may prove possible, in time, to rescue WAS cells using more conventional drug therapies.     

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ObjectivesVitronectin (VTN) has been widely used for the maintenance and expansion of human pluripotent stem cells (hPSCs) as feeder‐free conditions. However, the effect of VTN on hPSC differentiation remains unclear. Here, we investigated the role of VTN in early haematopoietic development of hPSCs.Materials and MethodsA chemically defined monolayer system was applied to study the role of different matrix or basement membrane proteins in haematopoietic development of hPSCs. The role of integrin signalling in VTN‐mediated haematopoietic differentiation was investigated by integrin antagonists. Finally, small interfering RNA was used to knock down integrin gene expression in differentiated cells.ResultsWe found that the haematopoietic differentiation of hPSCs on VTN was far more efficient than that on Matrigel that is also often used for hPSC culture. VTN promoted the fate determination of endothelial‐haematopoietic lineage during mesoderm development to generate haemogenic endothelium (HE). Moreover, we demonstrated that the signals through αvβ3 and αvβ5 integrins were required for VTN‐promoted haematopoietic differentiation. Blocking αvβ3 and αvβ5 integrins by the integrin antagonists impaired the development of HE, but not endothelial‐to‐haematopoietic transition (EHT). Finally, both αvβ3 and αvβ5 were confirmed acting synergistically for early haematopoietic differentiation by knockdown the expression of αv, β3 or β5.ConclusionThe established VTN‐based monolayer system of haematopoietic differentiation of hPSCs presents a valuable platform for further investigating niche signals involved in human haematopoietic development.     

Inhibition of whole body Ca2+ uptake in fresh water teleosts, Channa punctatus and Cyprinus carpio in response to salmon calcitonin

In many fish, ultimobranchial-derived calcitonin (CT) has been shown to be a potent hypocalcemic regulator. But an equal number of studies failed to show any correlation between CT and plasma calcium levels. Again, in fish, where CT has been shown to function as a hypocalcemic hormone, the way this is brought about is not well known. While the regulatory role of many hormones e.g., stanniocalcin, pituitary-derived prolactin and cortisol on gill calcium (Ca2+) transport (GCAT) has been well established, very few studies have been done to examine the effects of CT on GCAT in fish. In the present study we examined the effects of synthetic salmon calcitonin (sCT) in vivo on GCAT in two distinctly different species of fresh water teleost, Channa punctatus (partially air breathing) and Cyprinus carpio (fully gill breathing). Whole body calcium uptake, a measure of GCAT, was lower in the partial air breathing fish. We found that salmon CT had significant inhibitory effect on GCAT in both the fish species, kept either in normal tap water or low-calcium water. Fish, kept in high-calcium water, showed little response. In parallel studies we also observed that inhibition of GCAT was correlated with simultaneous changes in plasma calcium levels in response to exogenous administration of sCT. The present findings therefore suggest that CT in fresh water teleosts regulate its hypocalcemic action through inhibition of GCAT.     

Carbohydrase activity in the digestive system of some teleost fishes.

The activity of carbohydrases in Puntius sophore (Ham.), Channa gachua (Ham.) and Cirrhinus mrigala (Ham.) has been studied. The carbohydrases have been found in the stomach, intestinal bulb, intestine, pyloric caeca and the hepato-pancreas. The hepatopancreas is the main site of production of these enzymes and it is in this organ and the intestine that their activity is highest. Their pH optimum lies between 5.4 and 6.4. The enzyme equipment in the teleost is adapted to their respective food and feeding habits both qualitatively and quantitatively. In Puntius (omnivorous) and Cirrhinus (herbivorous) all three carbohydrases, namely amylase (EC 3.2.1.1.), sucrase (EC 3.2.1.26.) and raffinase, while in Channa (carnivorous) only amylase and sucrase have been found to be active. In Cirrhinus mrigala, which is predominantly a herbivorous species, the concentration of carbohydrases is higher than those in the other two fishes.