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Histochemical Methods (histochemical + methods)
Selected AbstractsRole Of Nitric Oxide On Pressor Mechanisms Within The Dorsomedial And Rostral Ventrolateral Medulla In Anaesthetized CatsCLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 3 2001Shu-Ying Chen SUMMARY 1. The role of nitric oxide (NO) in central cardiovascular regulation and the correlation between NO and glutamate-induced mechanisms is not clear. Microinjection of glutamate (3 nmol/ 30 nL) into dorsomedial medulla (DM) and rostral ventrolateral medulla (RVLM) increased arterial blood pressure (BP) and sympathetic vertebral nerve activity (VNA). Thus, in the present study, we examined the modulation by NO of glutamate-induced pressor responses in the DM and RVLM of cats. 2. Histochemical methods using nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) as a marker to stain neurons containing NO synthase (NOS), showed positive findings of NOS in both the DM and RVLM. 3. Microinjection of NG -nitro- L -arginine methyl ester (L -NAME), a NOS inhibitor, into the DM or RVLM did not alter resting BP and VNA, but it did cause a dose-dependent attenuation of glutamate-induced pressor responses. Interestingly, the increase in NO levels that resulted from pretreatment with L -arginine (L -Arg) or sodium nitroprusside (SNP) did not alter resting BP and VNA, but still inhibited glutamate-induced pressor responses in the DM and RVLM in a dose-dependent manner. 4. We also examined whether NO modulated the pressor responses induced by activation of different excitatory amino acid receptors. N -Methyl- D -aspartate (NMDA) and , -amino-3-hydroxy-5-methyl-4-isoxazole proprionic acid (AMPA) were used. Consistent with the results from the initial glutamate studies, we observed that not only L -NAME, but also L -Arg and SNP attenuated pressor responses induced by NMDA and AMPA. No difference was found between the effects of NO on NMDA- and AMPA-induced pressor responses. 5. To investigate the possibility of a loss of agonist selectivity, the effects of D -2-amino-5-phosphonovalerate (D -AP5) and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) on AMPA and NMDA responses in the DM were examined. The results showed that CNQX did not alter NMDA-induced pressor responses, while D -AP5 failed to alter AMPA-induced responses. 6. Our results suggest that activation of the glutamate-induced pressor mechanism is regulated by changes in NO levels in the DM and RVLM. This implies that NO may play a permissive role to allow operation of the glutamate-activation mechanism. [source] Localization of the A kinase anchoring protein AKAP79 in the human hippocampusEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 4 2000Attila Sík Abstract The phosphorylation state of the proteins, regulated by phosphatases and kinases, plays an important role in signal transduction and long-term changes in neuronal excitability. In neurons, cAMP-dependent protein kinase (PKA), protein kinase C (PKC) and calcineurin (CN) are attached to a scaffold protein, A kinase anchoring protein (AKAP), thought to anchor these three enzymes to specific sites of action. However, the localization of AKAP, and the predicted sites of linked phosphatase and kinase activities, are still unknown at the fine structural level. In the present study, we investigated the distribution of AKAP79 in the hippocampus from postmortem human brains and lobectomy samples from patients with intractable epilepsy, using preembedding immunoperoxidase and immunogold histochemical methods. AKAP79 was found in the CA1, presubicular and subicular regions, mostly in pyramidal cell dendrites, whereas pyramidal cells in the CA3, CA2 regions and dentate granule cells were negative both in postmortem and in surgical samples. In some epileptic cases, the dentate molecular layer and hilar interneurons also became immunoreactive. At the subcellular level, AKAP79 immunoreactivity was present in postsynaptic profiles near, but not attached to, the postsynaptic density of asymmetrical (presumed excitatory) synapses. We conclude that the spatial selectivity for the action of certain kinases and phosphatases regulating various ligand- and voltage-gated channels may be ensured by the selective presence of their anchoring protein, AKAP79, at the majority of glutamatergic synapses in the CA1, but not in the CA2/CA3 regions, suggesting profound differences in signal transduction and long-term synaptic plasticity between these regions of the human hippocampus. [source] Autophagic vacuolar myopathy in twin girlsNEUROPATHOLOGY & APPLIED NEUROBIOLOGY, Issue 3 2006J. L. Holton Hereditary autophagic vacuolar myopathy (AVM) may occur in several diseases including the rimmed vacuolar myopathies, acid maltase deficiency, Danon disease, infantile autophagic vacuolar myopathy and X-linked myopathy with excessive autophagy (XMEA). In the latter three conditions the vacuoles are lined by membranes with sarcolemmal features. We present two unusual cases of autophagic vacuolar myopathy in twin girls born at term with no family history of neurological disease. After initial normal developmental milestones they developed progressive leg weakness and wasting with contractures from the age of 12 years. Investigations showed raised CK, normal female karyotype, normal acid maltase activity, normal nerve conduction and myopathic EMG features. Frozen sections of skeletal muscle were stained using routine tinctorial and histochemical methods. Immunohistochemical staining for spectrin, merosin, dystrophin, complement membrane attack complex and sarcoglycans was performed and ultrastructural examination undertaken. Direct sequence analysis of the lamp-2 gene using genomic DNA extracted from lymphocytes was performed. Histological analysis of the muscle biopsies demonstrated myofibres with vacuoles lacking glycogen and lipid many of which were delineated using immunohistochemistry for merosin, dystrophin and sarcoglycans. Ultrastructural examination showed duplication of the myofibre basal lamina with associated autophagic material. Vacuoles within myofibres were either membrane bound containing autophagic material or lined by plasma membrane and basal lamina. Intermyofibrillar glycogen was increased. Sequence analysis of the coding region and intron/exon boundaries of the lamp-2 gene was normal. This is the first report of female cases of AVM with sarcolemmal features. We suggest that these patients may represent manifesting carriers of XMEA, or alternatively, a new form of disease with a similar phenotype having autosomal recessive inheritance. [source] Structure of the Oesophagus and Morphometric, Histochemical,Immunohistochemical Profiles of the Oesophageal Gland During the Post-hatching Period of Japanese Quails (Coturnix coturnix japonica)ANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 5 2009H. Sa Summary In the oesophagus, mucins, which originate from oesophageal submucosal glands, play an important role in the mucosal protection as a pre-epithelial barrier. In this study, the structure of cervical and thoracic parts of oesophagus of Japanese quail during the post-hatching period was compared, and the contents of carbohydrate and gastric mucin MUC5AC of the oesophageal glands in these parts were analysed at the light microscope levels by applying conventional histochemical and immunohistochemical methods. The oesophageal glands were present at hatching, located in the laminae propria. The numbers of glands were different in the cervical and thoracic parts, but the differences were found to be insignificant. The thoracic part has the oesophageal tonsils which are associated with the glands. Oesophageal tonsil was formed from day 5 after hatching. In quail of all ages, the secretory epithelium of glands contained neutral sialomucins and weakly sulphomucins. The cells in the neck region of secretory units contained sialomucins, while the cells of excretory ducts had strongly sulphomucins. Sialomucin containing cells in the secretory units increased with the advance of age and glandular development. But, in the secretory units, the sulphomucin content of glands was more in the thoracic part. The secretory epithelium of tonsil-associated glands contained mostly sulphomucins and a little sialomucin. From the hatching, MUC5AC mucin was detected in the cells of excretory ducts. Although the lymphoepithelium of the tonsil units exhibited negative reactions to all histochemical methods, it showed positive reaction to MUC5AC mucin antibody. In conclusion, the cervical and thoracic parts may be functionally different and the thoracic part of oesophagus was transformed into an immunological organ following day 5 after hatching. [source] Adrenergic and Cholinergic Innervation of the Mammary Gland in the PigANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 1 2002A. FRANKE-RADOWIECKA Adrenergic and acetylcholinesterase-positive (AChE-positive) innervation of the mammary gland in the sexually immature and mature pigs was studied using histochemical methods. Upon examining the adrenergic and cholinergic innervation, the adrenergic innervation was found to be much more developed. The majority of both sub-populations of the nerve fibres studied was localized in the subcutaneous tissue of the mammary gland. Adrenergic and AChE-positive nerve fibres also supplied structures of the nipple (subcutaneous tissue, blood vessels, smooth muscles fibres) and glandular tissue (blood vessels, lactiferous ducts). The glandular tissue contained the smallest number of adrenergic and AChE-positive nerve fibres. No distinct differences were observed in the adrenergic and AChE-positive innervation of the porcine mammary gland between the juvenile and non-pregnant adult animals. [source] IMPACT OF BLOOD FLOW OCCLUSION ON LIVER NECROSIS FOLLOWING THERMAL ABLATIONANZ JOURNAL OF SURGERY, Issue 1-2 2006Mehrdad Nikfarjam Background: Laser, radiofrequency and microwave are common techniques for local destruction of liver tumours by thermal ablation. The main limitation of thermal ablation treatment is the volume of necrosis that can be achieved. Blood flow occlusion is commonly advocated as an adjunct to thermal ablation to increase the volume of tissue necrosis based on macroscopic and histological assessment of immediate or direct thermal injury. This study examines the impact of blood flow occlusion on direct and indirect laser induced thermal liver injury in a murine model using histochemical methods to assess tissue vitality. Methods: Thermal ablation produced by neodymium yttrium-aluminium-garnet laser (wavelength 1064 nm) was applied to the liver of inbred male CBA strain mice at 2 W for 50 s (100 J). Treatment was performed with and without temporary portal vein and hepatic artery blood flow occlusion. Animals were killed upon completion of the procedure to assess direct thermal injury or at 24, 48 and 72 h to assess the progression of tissue damage. The maximum diameter of necrosis was assessed by vital staining for nicotinamide adenine dinucleotide (NADH) diaphorase. Microvascular changes were assessed by laser Doppler flowmetry, confocal in vivo microscopy and scanning electron microscopy. Results: The direct thermal injury (mean SE) assessed by NADH diaphorase staining was significantly greater following thermal ablation treatment without blood flow occlusion than with blood flow occlusion (3.3 (0.4) mm vs 2.9 (0.3) mm; P = 0.005). Tissue disruption, cracking and vacuolization was more pronounced adjacent to the fibre insertion site in the group treated with thermal ablation combined with blood flow occlusion. There was an equivalent increase in the extent of injury following therapy in both groups that reached a peak at 48 h. The maximum diameter of necrosis in the thermal ablation alone group at 48 h was significantly greater than the thermal ablation combined with blood flow occlusion group (5.8 (0.4) mm vs 5.3 (0.3) mm; P = 0.011). The patterns of microvascular injury were similar in both groups, varying in extent. Conclusion: Temporary blood flow inflow occlusion appears to decrease the extent of initial injury measured by vital staining techniques and does not alter the time sequence of progressive tissue injury following thermal ablation therapy. [source] Ontogenesis of internal secretory cells in leaves of Lantana camara (Verbenaceae)BOTANICAL JOURNAL OF THE LINNEAN SOCIETY, Issue 4 2005MARIA ZABELĘ DANTAS MOURA Internal secretory cells may be morphologically indistinguishable from their neighbours except for the presence of secreted material, or they may differ to such an extent that could be classified as secretory idioblasts. Several authors have reported the presence of glandular trichomes secreting essential oil in Verbenaceae, including Lantana. However, none have reported internal secretory cells. Anatomical and histochemical methods applied to Lantana camara leaves revealed the occurrence of internal secretory cells whose ontogenesis and chemical nature are described in this paper. According to leaf developmental analysis, L. camara secretory cells originated from the ground meristem, started to differentiate in the third node leaves, and were actively secreting in the fourth node leaves. The content of the secretory cells was of a lipidic nature, and a terpenoid essence of their secretion was also identified. Based upon differences in size and shape from neighbouring cells and on detection of nonvolatile terpenes, they were confirmed as true internal secretory idioblasts. © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society, 2005, 148, 427,431. [source] | |||||||||||||