Genesis,ultrastructure and cytochemical study of the cat eosinophil

Eosinophils from cat bone marrow and peripheral blood were studied by electron microscopy and cytochemical procedures. The maturation of eosinophils and formation of typical granules were described. Contrary to the accepted opinion that the core of animal's eosinophilic specific granules have a crystal-like structure, our observations revealed that the core has a myelin-like cylindrical appearance, whose layered formation proceeds from the inside outwards. Electron microscopic observations revealed that localization of reaction product to potassium pyroantimonate and phosphotungstic acid and to acid phosphatase activity was similar to that of eosinophils of man and other animals. Antimonate deposits and acid phosphatase activity were detected between the layers of the myelin-like structure of the core. Eosinophil granules failed to yield a positive reaction for peroxidase activity. The secretory activity of the eosinophil is discussed.     

Effect of lithium on the catecholamine concentration in the rabbit and rat brain

A single injection (200 mg/kg, intraperitoneally) or a course of injections (100 mg/kg subcutaneously, daily for 10 days) of lithium chloride given to rats had no significant effect on the content of catecholamines and dihydroxyphenylalanine in the brain stem 1, and 4 h after the injections. In experiments on rabbits the compound (100 mg/kg, intravenously) increased the noradrenalin concentration in the thalamus, hypothalamus, reticular formation, and caudate nucleus. An increase in the dopamine content in the caudate nucleus was accompanied by a simultaneous decrease in its concentration in the thalamus, hypothalamus, reticular formation, amygdala, and hippocampus.     

Induction of gastric histamine synthesis by H2-receptor antagonists: Potentiation of their antisecretory activity by histidine decarboxylase inhibitors

The activation of histamine (HA) formation in rat stomach was measured after repeated administrations of histamine H₂-receptor antagonists and the potentiation of their antisecretory activity was examined in histidine decarboxylase inhibitor (HDI)-pretreated rats. 40 mg/kg of metiamide, given intraperitoneally (i.p.) 24, 16 and 2 h prior to the examination, produced approximately 50% increase in the amount of¹⁴C-histamine, formed from¹⁴C-histidine in the stomach, and an almost equal enhancement in the gastric histidine decarboxylase (HD) activity. An equal dose of the compound did not influence the endogenous histamine level in the glandular stomach whereas it caused a significant increase in the serum histamine content. By similar treatment, 10 mg/kg of cimetidine enhanced the newly formed histamine in the rat stomach by 57%. The potent HDI, 2-hydroxy-5-carbomethoxy-benzyloxyamine (GYKI-11 121) suppressed the metiamide- and eimetidine-induced increases in histamine synthesis to slightly above or below the control values. In pharmacological studies, the antisecretory activity of histamine H₂-receptor blockers could markedly be potentiated by HDI. In GYKI-11 121 and NSD-1055-pretreated rats, the inhibiting potency of metiamide and cimetidine on pentagastrin-stimulated gastric acid secretion, increased to approximately twice that of the original effect. Neither GYKI-11 121 nor NSD-1055 produced significant inhibition on pentagastrin-stimulated gastric acid secretion in the applied doses. These findings provided evidence for the feedback stimulation of gastric HA synthesis by H₂-receptor blockers and confirmed the role of HA in the gastric acid secretion. Potentiation of the antisecretory activity of H₂-receptor antagonists by HDI would be useful in the therapeutic application of these compounds.     

The non-immunosuppressive immunophilin ligand GPI-1046 potently stimulates regenerating axon growth from adult mouse dorsal root ganglia cultured in Matrigel

We used explant cultures of adult mouse dorsal root ganglia with spinal nerve attached growing in Matrigel to assess the effects of the non-immunosuppressive immunophilin ligand GPI-1046 [Snyder et al. (1998) TIPS 19, 21-26] on the growth rate of regenerating sensory axons and found a potent stimulation of axon growth. In these explant cultures, naked, unfasciculated axons emerge from the cut end of the spinal nerve and continue to grow in the Matrigel for up to eight days [Tonge et al. (1996) Neuroscience 73, 541-551]. Some axons are entirely smooth whilst others show prominent varicosities. Some of the former express the phosphorylated neurofilament epitope recognised by monoclonal antibody RT97, a marker for large calibre, myelinated axons, whilst the latter express calcitonin gene-related peptide, predominantly a marker for unmyelinated, and small diameter myelinated sensory axons. Many of the axons in these cultures also express the low-affinity neurotrophin receptor p75. GPI-1046 has been shown to have striking stimulatory effects on embryonic primary sensory axons growing in vitro and it was therefore of interest to see whether it could also enhance regenerating sensory axon growth from the adult ganglia in our cultures. GPI-1046 potently stimulated axon growth in our cultures in a dose-dependent manner. The stimulatory effect was not dependent on the class of sensory axon. These observations show that GPI-1046 is a potent stimulator of regenerating axons from adult, primary sensory neurones. The cellular site of action of GPI-1046 is unknown. To distinguish between a direct effect of the drug on neurones and an indirect effect we compared the effects of GPI-1046 on explant and dissociated cultures. In confirmation of previous results, we found that GPI-1046 potently stimulated axon outgrowth from explants of embryonic chick dorsal root ganglia. However, the drug was without effect on dissociated embryonic dorsal root ganglion neurones, suggesting that non-neuronal cells are important for axon growth stimulation.     

Tapping allergen repertoires by advanced cloning technologies

BACKGROUND: Complex allergenic sources such as moulds, foods and mites contain complex panels of IgE-binding molecules which need to be cloned, produced and characterized in order to mimic the entire allergenicity of whole extracts reconstituted by mixing single standardized recombinant allergens. METHODS: Phage surface display of cDNA libraries selectively enriched for allergen-expressing clones using IgE from allergic patients allows rapid isolation of large panels of allergens. For the characterization of all different clones present in enriched cDNA libraries in a fast and cost-effective way, high-throughput screening technology is required. RESULTS: The combination of selective enrichment of cDNA libraries based on biopanning against serum IgE from sensitized patients and automated robot technology for picking and high-density gridding of clones onto filter membranes, followed by hybridization, enables fast identification of all the different clones present in an enriched library. The consequent application of selective enrichment and robotic-based screening allows, within weeks, cloning and characterization of the whole allergenic repertoire of any organisms. CONCLUSIONS: Robotic-based high-throughput screening of clones selected for IgE-binding capacity from phage surface-displayed cDNA libraries of Aspergillus fumigatus, Cladosporium herbarum, Coprinus comatus, Malassezia furfur, peanut and human lung tissue allowed rapid characterization of 81, 28, 37, 27, 8 and 151 different sequences, respectively. All these cDNAs bear a high probability to encode allergens derived from the respective allergenic source.     

Topography of corpuscular mechanoreceptors in the shoulder joint region of monodelphis domestica

The topography and structure of corpuscular mechanoreceptors in the shoulder joint capsule and periarticular connective tissue of a small laboratory marsupial (monodelphis domestica) were studied using light and electron microscopy. This animal is known to use its upper extremities for a wide range of activities like climbing and manipulating food. Thus, the shoulder joint of this animal species has a similar wide range of movement as the human shoulder joint, but is small enough for serial sectioning in its entirety. Silver stained serial paraffin sections were examined under the light microscope and the distribution of the different types of mechanoreceptors was reconstructed using three-dimensional image processing. In addition, selected mechanoreceptors were studied electron microscopically. Approximately 100 small lamellated corpuscles were found in the dense connective tissue of the joint capsule close to the insertion on the scapula and in the thickening of the joint capsule close to the glenoid labrum. Ruffini corpuscles were found in much smaller numbers in the moderately dense connective tissue of the axillary region. Only very few Vater-Pacinian corpuscles were seen in the soft periarticular connective tissue. The large number and localization of mechanoreceptor corpuscles in the shoulder joint capsule especially close to the glenoid labrum suggests, that these specialized nerve endings are likely to play an important role in control of joint movement. They can induce protective reflexes during extreme movements in the shoulder joint preventing shoulder luxation by increasing the tone of muscles pressing the humerus head into the glenoid cavity.     

Left Bochdalek hernia with delayed presentation: report of two cases

Left Bochdalek hernia is a serious and complex condition with high mortality. In most cases, it presents in the neonatal period and is seldom found later in life when symptomatology, usually after an asymptomatic period, is quite different and the prognosis excellent. The embryological development of left Bochdalek hernia suggests the presence of abdominal content in the left pleural cavity before birth. The type of clinical presentation and the prognosis depend on the time of visceral herniation. This study presents two cases of left Bochdalek hernia with delayed presentation. In both cases, after surgical reposition of the hernia, a small congenital diaphragmatic defect was found hidden between the rims of diaphragm. This indicates the possibility for the abdominal content to enter the left pleural cavity at the time of presentation.     

Direct comparison of umbilical cord blood versus bone marrow-derived endothelial precursor cells in mediating neovascularization in response to vascular ischemia.

Endothelial precursor cells (EPCs) cultured from adult bone marrow (BM) have been shown to mediate neovasculogenesis in murine models of vascular injury. We sought to directly compare umbilical cord blood (UCB)- and BM-derived EPC surface phenotypes and in vivo functional capacity. UCB and BM EPCs derived from mononuclear cells (MNC) were phenotyped by surface staining for expression of stromal (Stro-1, CXCR4, CD105, and CD73), endothelial (CD31, CD146, and vascular endothelial [VE]-cadherin), stem cell (CD34 and CD133), and monocyte (CD14) surface markers and analyzed by flow cytometry. The nonobese diabetic/severe combined immunodeficiency murine model of hind-limb ischemia was used to analyze the potential of MNCs and culture-derived EPCs from UCB and BM to mediate neovasculogenesis. Histologic evaluation of the in vivo studies included capillary density as a measure of neovascularization. Surface CXCR4 expression was notably higher on UCB-derived EPCs (64.29%+/-7.41%) compared with BM (19.69%+/-5.49%; P=.021). Although the 2 sources of EPCs were comparable in expression of endothelial and monocyte markers, BM-derived EPCs contained higher proportions of cells expressing stromal cell markers (CD105 and CD73). Injection of UCB- or BM-derived EPCs resulted in significantly improved perfusion as measured by laser Doppler imaging at days 7 and 14 after femoral artery ligation in nonobese diabetic/severe combined immunodeficiency mice compared with controls (P<.05). Injection of uncultured MNCs from BM or UCB showed no significant difference from control mice (P=.119; P=.177). Tissue samples harvested from the lower calf muscle at day 28 demonstrated increased capillary densities in mice receiving BM- or UCB-derived EPCs. In conclusion, we found that UCB and BM-derived EPCs differ in CXCR4 expression and stromal surface markers but mediate equivalent neovasculogenesis in vivo as measured by Doppler flow and histologic analyses.     

Beta-carotene-containing preparation carinat inhibits lipid peroxidation and development of renal tumors in rats treated with chemical carcinogen

The effects of pretreatment with β-carotene-containing preparation carinat on the development of renal tumors in rats receiving single intravenous injection of chemical carcinogen 3-(1-α-L-arabinopyranosyl)-1-methyl-1-nitrosourea were studied. Fourteen months after carcinogen administration, the degree of lipid oxidation in rat kidneys 2.5-fold surpassed that in animals receiving carinat in a dose producingin vivo antioxidant effect. Carinat decreased the total number of induced tumors and the incidence of mesenchymal renal tumors and suppressed the development of multiple tumors. The accumulation of lipoperoxides in the kidneys during carcinogenesis is associated with activation of free radical processes and carcinogen-induced inhibition of lipoperoxide enzymatic degradation and probably promotes renal malignancies due to co-carcinogenic action of these compounds. The data suggest that carinat-induced suppression of tumor development attests to antioxidant effects of β-carotene. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 130, No. 7, pp. 95–97, July, 2000