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Enterotoxaemia in sheep due to Clostridium welchii type D was indicated by field and laboratory investigations in Nepal. Morphological, cultural, biochemical, biological and toxin-producing characteristics observed were used to type the isolates. In anaerobic meat medium, all isolates produced pinkish discoloration of meat. All the strains fermented lactose, maltose, dextrose and sucrose whereas, salicine was fermented only by 17 strains. All but five strains were MR negative. Out of 200 isolated, 166 produced both alpha and epsilon toxins and the remaining 34 non-toxogenic strains are likely to be variants which have lost their toxogenicity. Epidemiologically the local name "Six months disease" and enterotoxaemia are considered to be identical diseases.
Vet Rec 1976 Jan 17
PMID:Initial studies on "six months disease" in sheep. 17 66

The isolectin B4 of Griffonia simplicifolia (GSA I-B4) binds to cell membrane glycoconjugates bearing terminal alpha-D-galactose, which macrophages possess. We have investigated the merits of its use as a marker for cells of this lineage when examining the early origin of macrophage populations in rat embryos, the stages and time scale of transformation from precursor forms to active, matured cells, and the response of precursors and macrophages to colony-stimulating blood factors, the last two studies conducted in organ cultures of prenatal lungs. In the present instance, GSA I-B4 was used either coupled with fluorescein (FITC) for light microscopy of living and fixed cells, or with peroxidase for light or electron microscopy. Control incubations of lung culture-derived macrophages proved that staining resulted from specific binding to galactosyl units on the cell membrane, since it was competitively inhibited by alpha-D-galactose. The lectin binds to few cells in 14-day prenatal lung explants but to a great many macrophages that subsequently develop in the cultures, indicating that it can be relied on for quantitative studies on population growth; however, it is important to provide reagents with good access to the cells. Apart from macrophages and their precursors, virtually no cells in prenatal lung cultures bind this lectin. Granulocytes of adult blood are GSA positive, but they are not yet present in 14-day prenatal explants and do not develop subsequent to culturing; hence they are not a source of confusion for experimental studies using this system. Precursors of granulocytes begin to appear in rat embryos around day 13 and have GSA-positive cell membranes, but like definitive granulocytes they also have conspicuous peroxidase-positive lysosomal granules which serve to distinguish them from early macrophages, particularly when cells are studied at an ultrastructural level. With these objections cleared away, GSA I-B4 emerges as a valuable means to mark cells of the macrophage line, mature or immature.
Anat Rec 1992 Apr
PMID:Macrophage development: I. Rationale for using Griffonia simplicifolia isolectin B4 as a marker for the line. 137 95

Earliest origins of macrophage populations in the central nervous system, the liver, and the lungs were studied in rat embryos aged between 10.5-11 days and 14 days of gestation, based on light and electron microscopic identification of macrophages using peroxidase-coupled isolectin B4 of Griffonia simplicifolia (GSA I-B4), which recognizes alpha-D-galactose groups on the cell membrane. During embryonic life macrophages and their precursors are GSA I-B4-positive and generally bereft of peroxidase-positive granules. At 10.5 days the yolk sac and embryonic circulations have just become joined, the brain has five vesicles but nerve cells are little differentiated, the liver exists as a diverticulum of the gut with fingerlike extensions of hepatocytes, and the lungs as a laryngotracheal groove. Macrophages and/or their precursors occurred in small numbers in embryonic mesenchyme and blood vessels but showed no special affinity for either liver or lung rudiments. The developing brain was the first organ to be colonized, beginning on prenatal day 12. The liver followed between days 12 and 13 and was succeeded by the lungs, beginning between days 13 and 14. Dividing macrophages were present in these organs at the outset of colonization and throughout the duration of the embryo series, indicating that from the beginning, replication of resident cells contributes to growth of the local population. Granulocyte precursors were first apparent in the liver around day 13; they are also GSA-positive but are distinguished from macrophages by their content of peroxidase-positive granules. Organ cultures of 13-day liver and lungs, and 14-day brain tissue, indicate that whereas isolated liver fragments support the formation of both granulocytes and macrophages, only the latter develop in brain or lung cultures. A resident population of macrophages evidently is set up very early in these organs, well before white cells colonize the spleen, bone marrow, and other future blood forming regions. The events outlined are seen as stages in an embryo-wide process that leads to establishment of macrophage populations in various organs.
Anat Rec 1992 Apr
PMID:Macrophage development: II. Early ontogeny of macrophage populations in brain, liver, and lungs of rat embryos as revealed by a lectin marker. 155 4

A selection of lectins was used to investigate developmentally regulated changes in the distribution of cell surface oligosaccharides during the gastrulation and neurulation stages of early chick embryo development. Lectins from three specificity classes were used: glucose/mannose specificity (concanavalin A [Con A], Lens culinaris agglutinin [LCA], Pisum sativum agglutinin [PSA]); N-acetylglucosamine specificity (Lycopersicon esculentum agglutinin [LEA], wheat germ agglutinin [WGA], succinylated WGA [sWGA]); N-acetylgalactosamine/galactose specificity (Dolichos biflorus agglutinin [DBA], soybean agglutinin [SBA], Sophora japonica agglutinin [SJA], Bandeiraea (Griffonia) simplicifolia lectin I [BSL I], peanut agglutinin [PNA], Artocarpus integrifolia lectin [Jacalin], Ricinus communis agglutinin-1 [RCA-1], Erythrina cristagalli lectin [ECL]). At gastrulation stages, patterns of lectin binding could be distinguished in the epiblast, mesoderm, and endoderm cell layers. The primitive streak failed to bind any of the lectins, but LEA and WGA bound to the epiblast in regions lateral to the streak, indicating the loss of some glucosamine residues medially in preparation for the ingression movements of gastrulation. Several lectins showed marked binding to the mesoderm cells after their passage through the primitive streak; these were LCA, PSA, WGA, sWGA, BSL, and most particularly PNA. Therefore, the epithelial-mesenchymal transformation from epiblast to mesoderm at the primitive streak is accompanied by cell surface oligosaccharide changes in the epiblast and mesoderm that involve all classes of lectins including the PNA-binding sequence Gal beta 1-3GalNAc. Ultrastructurally, PNA was shown to bind extracellularly to matrix fibrils. Jacalin, having the same sugar specificity as PNA, but binding to serine/threonine linked chains rather than asparagine linked chains showed no binding to the mesoderm.(ABSTRACT TRUNCATED AT 250 WORDS)
Anat Rec 1991 Oct
PMID:Changes in glycoconjugate expression during early chick embryo development: a lectin-binding study. 174 24

Norepinephrine administration causes progressive hypertrophy of the mammalian heart as measured by myocardial mass. The purpose of this study was to determine the growth response of the myocardial tissue components as well as the myocardial cell itself to norepinephrine. Young, adult cats were given low doses of norepinephrine in dextrose or dextrose alone twice daily for 15 days. On day 16, there were no changes in the animals body weight, right ventricular systolic pressure, right ventricular end-diastolic pressure, heart rate, cardiac index, or blood pressure. However, the right ventricle/body weight, the left ventricle/body weight and the total heart weight/body weight were increased significantly in the norepinephrine treated animals. The increase was on the order of 40%. The cardiac muscle cell was also significantly increased in size and both the right and left ventricular cardiac muscle cells exhibited a dramatic increase in size as measured by cross sectional area. Upon stereological examination it was found that the amount of hypertrophy as seen in the cardiac muscle cells was paralleled by the hypertrophy seen in the other tissue components of the myocardium. The volume density of the muscle cells, the interstitial components, as well as the blood vessel compartment were identical in the control and in the norepinephrine-treated groups. In conclusion, this study demonstrates that the response of the myocardium to norepinephrine is similar to that seen in response to a volume overload rather than that seen in response to pressure overload.
Anat Rec 1991 Apr
PMID:Norepinephrine-induced cardiac hypertrophy of the cat heart. 182 54

We demonstrated here with the high resolution lectin-gold approach and quantitative analysis, changes of glycoconjugates in the hamster zona pellucida (ZP) during oocyte growth and development in the ovary and the oviduct. Glycoconjugates which contain N-acetyl-D-galactosamine as terminal sugar residues are absent in the ovary but are secreted by secretory cells in the oviduct and are added to the ZP of superovulated oocytes during oviductal transit. Glycoconjugates which carry sialic acid as terminal sugar residues appear to be acquired mainly from the ovary. The oviduct contributes little of this particular component to the ZP during the transit of oocytes in the oviduct. On the contrary D-galactose and N-acetylglucosamine associated glycoconjugates, added to the ZP in ovarian follicles, are also secreted by non-ciliated oviductal epithelial cells and these secretory products are transferred to the ZP in significant amount during passage of the oocyte through the oviduct. Lectin-gold labeling of the ZP of superovulated oocytes reveals homogeneous distribution of gold particles throughout the zona matrix. Thus, we conclude that the ZP of hamster superovulated oocytes consists of glycoconjugates that may derive from different origins. Deposition of ZP glycocomponents begins in the ovary. Similar and new glycoconjugates, secreted by oviductal non-ciliated secretory cells, are added to the ZP of oocytes during oviductal transit. At this stage the ZP is made up of a homogeneous matrix of glycoconjugates.
Anat Rec 1991 Jul
PMID:Changes of glycoconjugate contents of the zona pellucida during oocyte growth and development in the golden hamster: a quantitative cytochemical study. 186 9

Lectin histochemistry at the light microscope level was used to determine the distribution of sugar residues in secretory cells of the olfactory mucosae of salamander, hamster, and mouse. Differences in sugar composition and distribution of glycoconjugates found in sustentacular cells and acinar cells of Bowman's glands of these three animals were characterized. Oligosaccharides in secretory products of sustentacular cells in salamander olfactory mucosa contained sialic acid, galactose (Gal), N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), fucose, and mannose residues. Glycoconjugates of these cells lacked terminal galactosyl-beta-(1,3)N-acetylgalactose (Gal beta 1,3GalNAc) residues. The sequences Gal beta 1,3GalNAc, N-acetyllactosamine (Gal beta 1,4GlcNAc), and GalNAc were penultimate to sialic acid residues. Sustentacular cells of mouse and hamster did not appear to contain O-linked oligosaccharides but stained for mannose-containing N-linked oligosaccharides. Glycoconjugates of acinar and duct cells of Bowman's glands in the salamander, hamster, and mouse contained variable amounts of beta(1,4)GlcNAc residues, and terminal N-acetyllactosamine, Gal beta 1,3GalNAc, and GalNAc residues. In the salamander, glycoconjugates of acinar cells possessed terminal GlcNAc residues but were not sialylated, while those of hamster and mouse generally stained for sialic acid but did not possess terminal GlcNAc residues. Secretory products of a subpopulation of rodent acinar cells also contained penultimate Gal beta 1,3GalNAc residues. Staining for sialic acid, Gal, GalNAc, and GlcNAc in glycoconjugates of rodents was often limited to a sub-population of Bowman's glands. This was especially noticeable in the mouse.
Anat Rec 1991 Apr
PMID:Identification of sugar residues in secretory glycoconjugates of olfactory mucosae using lectin histochemistry. 204 57

The binding of a panel of blood group-reactive lectins to frozen sections of human kidney was studied with a special emphasis on reactivity with endothelia and basement membranes. The blood group A-reactive lectins, all specific for alpha-D-N-acetylgalactosamine (GalNAc), Helix aspersa (HAA), Helix pomatia (HPA), and Griffonia simplicifolia I-A4 (GSA-I-A4) agglutinins bound to the endothelium in specimens with blood groups A and AB. In other samples, these lectins reacted predominantly with tubular basement membranes, as well as with certain tubules. Both Dolichos biflorus (DBA) and Vicia villosa agglutinins (VVA), reported to react with blood group A1 substance, failed to reveal endothelia in most specimens, but bound differently to tubules in all blood groups. The blood group B-reactive lectins, specific for alpha-D-galactose (alpha-Gal) or GalNAc, respectively, GSA-I-B4 and Sophora japonica agglutinin (SJA), bound to the endothelia in specimens from blood group B or AB and in other specimens bound only to certain tubules. Among the blood group O-reactive lectins, specific for alpha-L-fucose (Fuc), Ulex europaeus I agglutinin (UEA-I) conjugates, but not other lectins with a similar nominal specificity, bound strongly to endothelia in specimens with blood group O. The UEA-I conjugates bound distinctly more faintly to endothelia in specimens of other blood groups. The present results indicate that lectins, binding to defined blood group determinants, react with endothelia in specimens of the respective blood group status. Furthermore, they suggest that basement membranes and some tubules in the human kidney show a distinct heterogeneity in their expression of saccharide residues, related to their blood group status.
Anat Rec 1990 Jan
PMID:Binding of the blood group-reactive lectins to human adult kidney specimens. 229 76

Sheep and goat binucleate cells (BNC) play a central role in placental growth and development. This study reports a simple method for isolating 60-70% pure populations of BNC of high viability. After incubation of the isolated BNC with a brief pulse of 14C-leucine or 3H-fucose or 3H-galactose, electron microscope autoradiography showed that label was eventually incorporated into the characteristic BNC granules via the Golgi body. Fucose and galactose initially showed a much higher Golgi body label than leucine, which was at first predominantly localised in the endoplasmic reticulum. 35S-methionine incorporation by BNC suspensions was extensive enough to allow an immunoprecipitation investigation which demonstrated that the protein hormone ovine placental lactogen and the SBU-3 antigen were synthesised de novo. Previous studies with isolated BNC have shown a remarkable range of substances to be released into the incubation medium but not necessarily synthesised during the incubation. The results demonstrate unequivocally that isolated BNC's are capable of total synthesis in vitro of two of the proteins that these same cells are known to secrete in vivo.
Anat Rec 1990 Jan
PMID:Characterization of the synthetic capacities of isolated placental binucleate cells from sheep and goats. 229 81

The formation of acellular cementum and the deposition of [3H]mannose-labeled extracellular matrix were studied in 14-day-old Sprague-Dawley rats. The sequential events of cementogenesis and periodontal ligament formation observed by light and electron microscopy were described from the stage of an intact root sheath to postcementogenesis. Ultrastructural examination of cementoblasts and periodontal ligament fibroblasts revealed [3H]mannose labeling of the Golgi apparatus at 10 minutes, collagen secretion granules at 30 minutes, and the extracellular matrix beginning at 30 minutes. The extracellular matrix between cementoblasts and dentin was heavily labeled at 1 and 4 hours. Newly formed principal fibers of the periodontal ligament were also heavily labeled at 4 hours. Fully differentiated cementoblasts exhibited the largest sectional profiles and the highest number of silver grains per unit area of cytoplasm. The morphologic and radioautographic data suggest that during the formation of acellular cementum, the cementoblast phenotype is expressed for a short period of time, after which cementoblasts appear to mix with the fibroblasts of the periodontal ligament.
Anat Rec 1989 Feb
PMID:Radioautographic study of [3H]mannose utilization during cementoblast differentiation, formation of acellular cementum, and development of periodontal ligament principal fibers. 271 47


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