A Study on Nursing Students' Knowledge, Attitude, and Educational Needs for Brain-Death Organ Transplantation and Donation and Intent to Donate Organs.

OBJECTIVE: The purpose of this study was to identify the knowledge, attitude, educational needs, and will of nursing students on organ donation from brain-dead donors. METHODS: Data were collected by using a 40-item questionnaire to measure knowledge, attitude, educational needs, and will for organ donation of 215 nursing college students in one university in Dangjin city from May 11 to May 31, 2017. The data were analyzed using SPSS 22 program (Data Solution Inc, Seoul). RESULTS: In the general characteristics, 85.1% of the subjects did not receive education on donation, and 99.5% of the subjects responded that education is needed. The desired methods of education were special lecture in school (55.3%), "webtoons" on the Internet (19.5%), formal curriculum (15.8%). Points to improve to increase brain-death organ transplantation and donation included "active publicity through pan-national campaign activities" (56.3%), "respecting prior consent from brain-dead donors" (21.9%), and "encouragement and increased support for organ donors" (12.1%). There was a significant difference in knowledge according to will for organ donation (t = 3.29, P = .001) and consent to brain-death organ donation in family members (t = 3.29, P = .001). There was a statistically significant positive correlation between attitude and knowledge of the subjects regarding brain-death organ donation. CONCLUSION: The knowledge, attitude, educational need, and will for organ donation of nursing students revealed in this study will be used as basic data to provide systematic transplant education including contents about organ transplantation in the regular nursing curriculum in the future. It will contribute to the activation of organ donation.

Doxycycline treatment attenuates acute lung injury in mice infected with virulent influenza H3N2 virus: involvement of matrix metalloproteinases.

Acute respiratory distress syndrome, a severe form of acute lung injury (ALI), is a major cause of death during influenza pneumonia. We have provided evidence for the involvement of recruited neutrophils, their toxic enzymes such as myeloperoxidase and matrix metalloproteinases (MMPs), and neutrophil extracellular traps in aggravating alveolar-capillary damage. In this study, we investigated the effects of doxycycline (DOX), an inhibitor of MMPs, on influenza-induced ALI. BALB/c mice were infected with a sublethal dose of mouse-adapted virulent influenza A/Aichi/2/68 (H3N2) virus, and administered daily with 20mg/kg or 60 mg/kg DOX orally. The effects of DOX on ALI were determined by measuring inflammation, capillary leakage, and MMP activities. Furthermore, levels of T1-α (a membrane protein of alveolar type I epithelium) and thrombomodulin (an endothelial protein) in the bronchoalveolar lavage fluid were evaluated by Western blot analysis. Our results demonstrate significantly decreased inflammation and protein leakage in the lungs after DOX treatment. Levels of MMP-2 and MMP-9 activity, T1-α and thrombomodulin were also diminished in the DOX-treated group. These findings were corroborated by histopathologic analyses, which demonstrated significant reduction in lung damage. Although DOX treatment reduced ALI, there were no effects on virus titers and body weights. Taken together, these results demonstrate that DOX may be useful in ameliorating ALI during influenza pneumonia. Further studies are warranted to determine whether DOX can be used in combination with anti-viral agents to alleviate severe influenza pneumonia.

Adaptation of human influenza H3N2 virus in a mouse pneumonitis model: insights into viral virulence, tissue tropism and host pathogenesis.

Most pandemic influenza virus strains undergo adaptation or reassortment before they acquire the ability to cause fatal infections in a new host species. The pathologic changes and tissue tropism during virus adaptation are not fully understood. Here we investigated pathologic changes and tissue tropism by serial lung-to-lung passaging of human influenza virus strain A/Aichi/2/68 (H3N2) in a BALB/c mouse model. Enhanced pulmonary lesions and systemic virus infection were observed during adaptation. Late passage 10 (P10) virus caused extra-pulmonary spread with necrotic and inflammatory lesions in the brain, heart, spleen and intestine of infected animals, in contrast to infection with earlier passage viruses which were restricted to lungs. Non-conservative mutations in the hemagglutinin (Gly218Glu) and non-structural 1 (Asp125Gly) proteins were identified in P10 virus which exhibited high virulence. Virus growth kinetics showed enhanced replication ability of P10 virus in different cell lines. P10 virus also exhibited the ability to bind to erythrocytes of different host species. These results demonstrate extra-pulmonary spread of influenza virus during adaptation with enhanced replication ability in a new host. This mouse adaptation model may provide a basis for understanding cross-species adaptability corresponding to increased virulence of the influenza A virus, a phenomenon of relevance to the emergence of future highly pathogenic strains.

Involvement of AT(1) angiotensin receptors in the vasomodulatory effect of des-aspartate-angiotensin I in the rat renal vasculature.

Angiotensin II is known to act primarily on the angiotensin AT(1) receptors to mediate its physiological and pathological actions. Des-aspartate-angiotensin I (DAA-I) is a bioactive angiotensin peptide and have been shown to have contrasting vascular actions to angiotensin II. Previous work in this laboratory has demonstrated an overwhelming vasodepressor modulation on angiotensin II-induced vasoconstriction by DAA-I. The present study investigated the involvement of the AT(1) receptor in the actions of DAA-I on angiotensin II-induced vascular actions in the renal vasculature of normotensive Wistar-Kyoto rats (WKY), spontaneously hypertensive rats (SHR) and streptozotocin (STZ)-induced diabetic rats. The findings revealed that the angiotensin receptor in rat kidney homogenate was mainly of the AT(1) subtype. The AT(1) receptor density was significantly higher in the kidney of the SHR. The increase in AT(1) receptor density was also confirmed by RT-PCR and Western blot analysis. In contrast, AT(1) receptor density was significantly reduced in the kidney of the streptozotocin-induced diabetic rat. Perfusion with 10(-9)M DAA-I reduced the AT(1) receptor density in the kidneys of WKY and SHR rats suggesting that the previously observed vasodepressor modulation of the nonapeptide could be due to down-regulation or internalization of AT(1) receptors. RT-PCR and Western blot analysis showed no significant changes in the content of AT(1) receptor mRNA and protein. This supports the suggestion that DAA-I causes internalization of AT(1) receptors. In the streptozotocin-induced diabetic rat, no significant changes in renal AT(1) receptor density and expression were seen when its kidneys were similarly perfused with DAA-I.

Effect of des-aspartate-angiotensin I on the actions of angiotensin II in the isolated renal and mesenteric vasculature of hypertensive and STZ-induced diabetic rats.

The present study investigated the action of des-aspartate-angiotensin I (DAA-I) on the pressor action of angiotensin II in the renal and mesenteric vasculature of WKY, SHR and streptozotocin (STZ)-induced diabetic rats. Angiotensin II-induced a dose-dependent pressor response in the renal vasculature. Compared to the WKY, the pressor response was enhanced in the SHR and reduced in the STZ-induced diabetic rat. DAA-I attenuated the angiotensin II pressor action in renal vasculature of WKY and SHR. The attenuation was observed for DAA-I concentration as low as 10(-18) M and was more prominent in SHR. However, the ability of DAA-I to reduce angiotensin II response was lost in the STZ-induced diabetic kidney. Instead, enhancement of angiotensin II pressor response was seen at the lower doses of the octapeptide. The effect of DAA-I was not inhibited by PD123319, an AT2 receptor antagonist, and indomethacin, a cyclo-oxygenase inhibitor in both WKY and SHR, indicating that its action was not mediated by angiotensin AT2 receptor and prostaglandins. The pressor responses to angiotensin II in mesenteric vascular bed were also dose-dependent but smaller in magnitude compared to the renal vasculature. The responses were significantly smaller in SHR but no significant difference was observed between STZ-induced diabetic and WKY rat. Similarly, PD123319 and indomethacin had no effect on the action of DAA-I. The findings reiterate a regulatory role for DAA-I in vascular bed of the kidney and mesentery. By being active at circulating level, DAA-I subserves a physiological role. This function appears to be present in animals with diseased state of hypertension and diabetes. It is likely that DAA-I functions are modified to accommodate the ongoing vascular remodeling.

Metabotropic glutamate receptor 2/3 in the hippocampus of patients with mesial temporal lobe epilepsy, and of rats and mice after pilocarpine-induced status epilepticus.

A comparative study of the expression of metabotropic glutamate receptor 2/3 (mGluR2/3) was done in the hippocampus of rats and mice after pilocarpine-induced status epilepticus (APISE), and of patients with mesial temporal lobe epilepsy. At 1 day APISE, there was a marked increase in mGluR2/3 immunoreactivity in the stratum lacunosum moleculare (SLM) of CA1 area and in the middle one-third of the molecular layer (MM) of the dentate gyrus. Immuno-electron microscopic study showed degenerating mGluR2/3 positive axons in the SLM of CA1 area at 1 day APISE. From 7 days, mGluR2/3 immunopositive product decreased, and by 31 days APISE, it almost disappeared in two-thirds of the SLM near CA2. In the mouse model at 2 months APISE, mGluR2/3 immunopositive product in two-thirds of the SLM near the stratum radiatum disappeared, and so did in the whole SLM of CA1 area in patients with mesial temporal lobe epilepsy. Neuropharmacological study by intravenous injection of mGluR2/3 agonist 2R,4R-4-aminopyrrolidine-2,4-dicarboxylate [(2R,4R)-APDC] at different doses at 1h during pilocarpine induced status epilepticus showed that (2R,4R)-APDC could not stop seizures and neuronal death in the hilus of the dentate gyrus. The present study, therefore, suggests that the reduction of mGluR2/3 immunopositive product in the SLM of CA1 is a consequence of neuronal loss in either the entorhinal cortex or CA1 area of the hippocampus, and at the dosage range from 12.5 to 600 mg/kg, (2R,4R)-APDC may not be effective in the prevention of seizures or neuronal death in the hilus of the dentate gyrus.

Effects of des-aspartate-angiotensin I on the actions of angiotensin III in the renal and mesenteric vasculature of normo- and hypertensive rats.

An earlier study showed that des-aspartate-angiotensin I (DAA-I) attenuated the pressor action of angiotensin III in aortic rings of the spontaneously hypertensive rat (SHR) but not the normotensive Wistar Kyoto (WKY) rat. The present study investigated similar properties of DAA-I in isolated perfused kidneys and mesenteric beds of WKY and SHR. In the renal vasculature, angiotensin III induced a dose-dependent pressor response, which was more marked in the SHR than WKY in terms of significant greater magnitude of response and lower threshold. DAA-I attenuated the pressor action of angiotensin III in both the WKY and SHR. The attenuation in SHR was much more marked, occurring at doses as low as 10(-15) M DAA-I, while effective attenuation was only seen with 10(-9) M in WKY. The effects of DAA-I was not inhibited by PD123319 and indomethacin, indicating that its action was not mediated by angiotensin AT2 receptors and prostaglandins. However, the direct pressor action of angiotensin III in the SHR but not the WKY was attenuated by indomethacin suggesting that this notable difference could be due to known decreased response of renal vasculature to vasodilator prostaglandins in the SHR. Pressor responses to angiotensin III in the mesenteric vascular bed was also dose dependent, but smaller in magnitude compared to the renal response. The responses in the SHR, though generally smaller, were not significantly different from those of the WKY. This trend is in line with the similar observations with angiotensin III and II by other investigators. In terms of the effect of DAA-I, indomethacin and PD123319 on angiotensin III action, similar patterns to those of the renal vasculature were observed. This reaffirms that in the perfused kidney and mesenteric bed, where the majority of the vessels are contractile, femtomolar concentrations of DAA-I attenuates the pressor action of angiotensin III. The attenuation is not indomethacin sensitive and does not involve the angiotensin AT2 receptor. The findings suggest that DAA-I possesses protective vascular actions and is involved in the pathophysiology of hypertension.

Effects of des-aspartate-angiotensin I on the expression of angiotensin AT1 and AT2 receptors in ventricles of hypertrophic rat hearts.

The effects of des-aspartate-angiotensin I (DAA-I) on the expression of angiotensin AT1 and AT2 receptor in hearts of aortic coarcted rats were studied. The protocols used included competitive reverse transcription polymerase chain reaction (RT-PCR), Western blotting, and receptor-ligand binding assays. mRNA of the AT1 and AT2 receptors increased significantly after 4 days of aortic coarctation (7- and 4-folds of sham-operated, respectively). However, the protein of the AT1 receptor was not altered, and only increase in protein of the AT2 receptor was detected. There was an increase in [125I]Sar1-Ile8-angiotensin II binding sites in the ventricular membranes of hypertrophic hearts, which was attributed to an upregulation of the AT2 receptor. Treatment with i.p. DAA-I resulted in a significant reduction of cardiac hypertrophy, the maximum effect was achieved with a dose of 200 nmol/kg/day. The anti-cardiac hypertrophy effect appeared to be U-shape, and at a higher dose of 800 mol/kg/day, there was a loss of effect. DAA-I had no effect on the receptor protein in ventricles of hypertrophic hearts. However, DAA-I dose-dependently decreased the binding of [125I]Sar1-Ile8-angiotensin II to ventricular membranes. The decrease was due to a likely desensitization by internalization of the AT1 receptor, and this probably contributed to the loss of hypertrophic effects at 800 nmol/kg/day. Treatment of DAA-I also resulted in a remarkable increase in AT2 receptor mRNA (24-fold increase over the sham-operated), which was not coupled to translation. The present findings provide new information regarding the relationship between cardiac hypertrophy and the angiotensin receptors, and the anti-cardiac hypertrophic actions of DAA-I via the AT1 receptors.

Angiotensins in plasma of hypertensive rats and human.

The plasma levels of des-aspartate-angiotensin I (DAA-I) in three models of hypertensive rats and hypertensive subjects were determined and compared with their normotensive controls. The rationale for the study was based on our earlier findings showing that DAA-I is a physiological angiotensin peptide that is involved in the pathophysiology of the cardiovascular system. The determination was carried out by the technique of capillary electrophoresis. Plasma level of angiotensin I, angiotensin II, and angiotensin III was also determined as a measurement of the status of the renin-angiotensin system in the different models of hypertension. DAA-I was found to be significantly lower in the spontaneously hypertensive rats (SHR) (46.6 +/- 2.5 pmol/l compared to 66.1 +/- 3.4 pmol/l for the normotensive control Wistar Kyoto rats), renal hypertensive rats (54.2 +/- 5.1 pmol/l compared to 72 +/- 2.5 pmol/l for the normotensive control Sprague-Dawley rats), and essential human hypertensive subjects (15.2 +/- 0.9 pmol/l compared to 19.5 +/- 2.5 pmol/l for the normotensive adult), whilst plasma concentration of angiotensin I and angiotensin II is reflective of the state of the renin-angiotensin system in the particular model of hypertension. When the SHR and human hypertensive subjects were treated with an angiotensin converting enzyme (ACE) inhibitor, the plasma level of DAA-I increased significantly. These findings suggest that the low plasma level of DAA-I could be a characteristic defect of the renin-angiotensin system in the two genetic models of hypertension (SHR and human essential hypertensive subjects). The increase of the nonapeptide following ACE inhibitor treatment could be an important hitherto unrecorded contributory factor to the effectiveness of ACE inhibitors in combating heart pathology.

Location of renin-angiotensin system components in the hypoglossal nucleus of the rat.

The renin-angiotensin system (RAS) in the hypoglossal nuclei of the rat was studied by immunohistochemistry. Antibodies to angiotensin AT(1) receptor (AT1), angiotensinogen (ANG), renin (REN), angiotensin converting enzyme (ACE) and angiotensin II (AII) were used. All the components of the RAS with the exception of renin were detected. Light and electron microscopy revealed the following results: ANG was predominantly found in astrocytes, with small amounts in neuronal dendrites; ACE was found in the cytoplasm of neurons, dendrites and astrocyte processes; AT1 was found in the cytoplasm of neurons and dendrites, but not on the membrane; and AII was found mainly in astrocytes with some located in the dendrites and cytoplasm. Right hypoglossal nerve lesion caused an increase in expression of AT1 in neurons as early as 2 days post-lesion. An increase in expression of ANG in astrocytes was also seen, but at a much later time of 3 weeks post-lesion. For AII, staining occurred in both the neurons and astrocytes in the undamaged hypoglossal nucleus. Nerve lesion caused a disappearance of neuronal stains and an increase in astrocyte stains. There were no changes in ACE staining after nerve lesion. We speculate that ANG and AII are made within the astrocytes, whereas ACE could either be uptaken from blood or de novo synthesized. AT1 may potentially be internal soluble receptors. As to the function of AII in the hypoglossal nucleus, the data do not support AII as a neurotransmitter in the hypoglossal nucleus. It may function as a neuromodulator and also be involved in basic cellular activities, e.g. regulation of transcription factors.

Expression of metabotropic glutamate receptor 1alpha in the hippocampus of rat pilocarpine model of status epilepticus.

The expression of metabotropic glutamate receptor 1alpha was studied in the rat hippocampus after pilocarpine-induced status epilepticus by Western blot and immunocytochemistry at both light and electron microscopic levels. At 1 day after pilocarpine-induced status epilepticus, there was marked decrease in metabotropic glutamate receptor 1alpha immunoreactivity at the border between stratum oriens and alveus in CA1 and CA3, and in the hilus of dentate gyrus. Between 3 and 31 days after pilocarpine-induced status epilepticus, metabotropic glutamate receptor 1alpha-immunoreactive dendrites and cell bodies in the border between stratum oriens and alveus gradually reappeared. Upregulation of metabotropic glutamate receptor 1alpha, however, was observed in the stratum oriens of CA1 at day 1, but returned to baseline by day 7. By electron microscopy, the metabotropic glutamate receptor 1alpha-immunoreactive product was demonstrated only in the post-synaptic elements in the border between the stratum oriens and alveus of CA1 and the hilus of the dentate gyrus in both control and experimental rats. At 1 day after pilocarpine-induced status epilepticus, metabotropic glutamate receptor 1alpha-immunoreactive degenerating neurons were identified in the border between stratum oriens and alveus of CA1 and the hilus of the dentate gyrus. At 7 and 31 days, many degenerating axons were also found. Present results suggest that excitoneurotoxicity mediated through post-synaptic metabotropic glutamate receptor 1alpha may be involved in degeneration and death of interneurons in the hilus of dentate gyrus, and the border between stratum oriens and alveus of CA1 in the early stage after pilocarpine-induced status epilepticus.

Cardiovascular actions of chicken-meat extract in normo- and hypertensive rats.

The cardiovascular actions of a commercial chicken-meat extract known as Brand's Essence of Chicken (Cerebos Pacific Ltd, Singapore; BEC) were investigated in normo- and hypertensive rats. The spontaneously-hypertensive rat (SHR), Wistar Kyoto rat (WKY) and Sprague Dawley rat (SD) were used. The effect of oral feeding of BEC on hypertension, cardiac hypertrophy and arteriosclerosis in these animals was studied. The data showed the following effects of oral feeding of BEC: (1) feeding for 30 d did not affect the blood pressure and heart rate (determined telemetrically) of adult SHR and WKY; (2) feeding for 90 d did not affect the development of hypertension in 1-month-old prehypertensive SHR; (3) feeding for 4 d dose-dependently (0.2--3.2 ml/kg per d) attenuated cardiac hypertrophy in experimentally-induced (coarctation of the abdominal aorta) cardiac hypertrophic SD; (4) feeding to 1-month-old prehypertensive SHR for 11 months did not affect the age-related development of hypertension in this animal; (5) there was significant attenuation of the age-related development of hypertension (determined by tail-cuff plethysmography) in the WKY (P = 0.011) when the animals drank an average of 7.5 ml BEC/kg body weight per d, measured during the last 2 months of the 11-month treatment period; (6) there was chronic, as in the previous treatment, attenuation of the age-related development of cardiac hypertrophy and arteriosclerosis (quantified morphometrically) in the SHR when the animals drank an average of 2.4 ml BEC/kg per d, measured during the last 2 months of the 11-month treatment period. A parallel study using laboratory-prepared chicken-meat and pork extracts showed that the former, but not the latter, attenuated cardiac hypertrophy in experimentally-induced cardiac hypertrophic SD. These findings, showing that chicken-meat extract (both BEC and laboratory prepared) could have anti-cardiac hypertrophic, anti-hypertensive and anti-arteriosclerotic actions, were unexpected and provoking, and would challenge nutritional scientists with an interest in meat consumption and cardiovascular diseases.

Metabotropic glutamate receptor 8 in the rat hippocampus after pilocarpine induced status epilepticus.

The expression of metabotropic glutamate receptor 8 (mGluR8) was studied in the rat hippocampus after pilocarpine-induced status epilepticus (APISE) by light immunohistochemistry and immunoelectron microscopy. At 1 day APISE, mGluR8 immunoreactivity was up-regulated in the entire molecular layer of the dentate gyrus. At 7 days APISE, mGluR8 immunoreactive cells began to appear in the stratum lacunosum moleculare of CA1, and by day 31, they were seen in all layers of CA1. By electron microscopy and double labelling study, the mGluR8 immunoreactive cells were identified as astrocytes. The present novel finding of induced expression of mGluR8 in astrocytes APISE suggests that it may be linked to gliosis.

Effects of des-aspartate-angiotensin I on angiotensin II-induced incorporation of phenylalanine and thymidine in cultured rat cardiomyocytes and aortic smooth muscle cells.

Des-aspartate-angiotensin I, a pharmacologically active nine-amino acid angiotensin peptide, and losartan, an AT(1) angiotensin receptor antagonist, but not angiotensin-(1-7), another active angiotensin peptide, completely attenuated the angiotensin II-induced incorporation of [3H]phenylalanine in cultured rat cardiomyocytes. The attenuation by des-aspartate-angiotensin I but not that of losartan was inhibited by indomethacin. The data support an earlier suggestion that the nonapeptide attenuates cardiac hypertrophy in rats via an indomethacin-sensitive angiotensin AT(1) receptor subtype. In rat aortic smooth muscle cells, both des-aspartate-angiotensin I and angiotensin-(1-7) had no effect on the angiotensin II-induced [3H]phenylalanine incorporation. However, the two peptides significantly attenuated the angiotensin II-induced [3H]thymidine incorporation in the smooth muscle cells. The attenuation by angiotensin-(1-7) but not by des-aspartate-angiotensin I was inhibited by (D-Ala(7))-angiotensin-(1-7), a specific angiotensin-(1-7) antagonist. Des-aspartate-angiotensin I also attenuated FCS-stimulated [3H]thymidine incorporation. This attenuation was inhibited by the peptide angiotensin receptor antagonist, (Sar(1), Ile(8))-angiotensin II, but not by losartan. These data indicate that des-aspartate-angiotensin I and angiotensin-(1-7) do not participate in the process of protein synthesis in vascular smooth muscle cells and that the nonapeptide and heptapeptide act on different non-AT(1) receptors to mediate their anti-hyperplasic action. Although the exact mechanisms of action remain to be elucidated, the findings indicate that des-aspartate-angiotensin I acts as an agonist on angiotensin AT(1) and non-AT(1) receptor subtypes and induces responses that oppose the actions of angiotensin II.

Acetylcholinesterase-independent action of diisopropyl-flurophosphate in the rat aorta.

Recent studies have shown that many organophosphates can bind competitively and noncompetitively to membrane muscarinic receptors. The present study investigated the responses of the rat aortic rings to diisopropyl-flurophosphate (DFP), an organophsophorus cholinesterase inhibitor, and the possible involvement of muscarinic receptors. DFP caused a concentration-dependent contraction when added cumulatively from 10(-8) to 10(-4) M. This contraction was inhibited in a noncompetitive manner by high concentrations of atropine (1.5 x 10(-6) and 1.8 x 10(-6) M) but was unaffected by similar concentrations of selective muscarinic receptor subtype antagonists, pirenzepine, 11-2[2-[(diethylamino)methyl]-1-piperidinyl]acetyl-5, 11-dihydro-6H-rido[2,3-b][1,4]benzodiazepin-6-one (AF-DX116) and 4-Diphenylacetoxy-N-methyl piperidine methiodide (4-DAMP). Phentolamine, an alpha-adrenoceptor antagonist, was able to inhibit the DFP-induced contraction in a noncompetitive manner at a concentration of 10(-7) M. These findings suggested that the DFP-induced contraction in the rat aortic rings was mediated by norepinephrine that was released from sympathetic nerve terminals present in the aortic rings.

Actions of angiotensin peptides on the rabbit pulmonary artery.

The presence of the angiotensin AT1A-like receptor subtype in the pulmonary artery and AT1B-like receptor subtype in the pulmonary trunk of the rabbit has been reported in two earlier studies. The present study further investigated these receptor subtypes using five other angiotensins (namely angiotensin II, angiotensin III, angiotensin IV, angiotensin-(1-7) and angiotensin-(4-8)). The direct action of the angiotensins on the rabbit pulmonary arterial and trunk sections and the ability of each angiotensin to further contract or relax preconstricted sections of the pulmonary artery and trunk were studied using the organ bath set-up. The effects of angiotensin III on the 3H overflow from re-uptaken [3H]noradrenaline in the electrically-contracted rabbit pulmonary arterial and trunk sections were also studied. The contractile response of the arterial and trunk section had the following rank order potency: angiotensin II > angiotensin III > angiotensin IV. The contractile response to these angiotensins was greatly reduced or absent in the pulmonary trunk. Angiotensin II further contracted the preconstricted arterial and trunk sections. In contrast, angiotensin III further contracted the preconstricted arterial section but relaxed the preconstricted trunk section. Angiotensin IV similarly relaxed the preconstricted trunk section but had minimum effect on the preconstricted arterial section. Angiotensin-(1-7) and angiotensin-(4-8) had no effect on both sections. The actions of the three angiotensins were inhibited by losartan, an AT1-selective antagonist. Indomethacin, a cyclo-oxygenase inhibitor, inhibited the relaxation caused by angiotensin III and angiotensin IV in the trunk section. The effects of angiotensin III on the electrically preconstricted sections of the pulmonary trunk and artery were not accompanied by any significant changes in 3H overflow. The differential responses produced by angiotensin II and its immediate metabolites via two positionally located and functionally opposing receptor subtypes suggest that the pulmonary trunk and artery is not a passive conduit but an important regulator of blood flow from the heart to the lung.

Pre- and/or post-synaptic localisation of metabotropic glutamate receptor 1alpha (mGluR1alpha) and 2/3 (mGluR2/3) in the rat spinal cord.

Using immunocytochemical techniques (light and electron microscopy), weakly stained metabotropic glutamate receptor (mGluR) 1alpha immunoreactivity was detected in lamina I of the rat spinal cord. Immunoreactivity for mGluR2/3 was almost undetectable in this lamina and outer lamina II. In lamina II, there was mGluR1alpha immunoreactivity. Strongly stained mGluR2/3 was seen in the inner layer of lamina II and the dorsal part of lamina III. In laminae III X, weakly to moderately stained mGluR1alpha immunoreactive product was demonstrated. Similar staining for mGluR2/3 was also seen in lamina III-VI and in lamina X, but mGluR2/3 immunoreactivities were few in lamina VII-IX. With electron microscopy, mGluR1alpha immunoreactivity was seen in neuronal cell body and dendrites in lamina II of the dorsal horn. In the lateral and ventral horns, only dendrites of neurons were mGluR1alpha immunopositive. Some mGluR2/3 immunopositive dendrites were demonstrated in lamina II of the dorsal horn, lateral and ventral horns. In the ventral horn, mGluR2/3 immunopositive axon and axon terminals were demonstrated. Some mGluR2/3 immunopositive astrocytes were also demonstrated in the three areas and their strongly stained processes wrapped around neuronal cell bodies and synapses.

M3/M1-Selective antimuscarinic tropinyl and piperidinyl esters.

The binding affinities of some tropinyl and piperidinyl esters for the submandibulary glands (M3/M1) and heart ventricle (M2) were determined from displacement experiments using 3H-labelled N-methylscopolamine as radioligand. The antimuscarinic activities of these esters were also evaluated on guinea pig bronchi. The esters inhibited the M3-mediated carbachol-induced contraction of the bronchial smooth muscle and a reasonable correlation was obtained between the binding affinities of the esters for the submandibulary glands (pKM3,M1) and their inhibitory activities (pIC50) on guinea pig bronchi. A promising compound, N-methylpiperidinyl cyclohexylphenylpropionate (NCPP) which combined good antimuscarinic activity (pA2=9.34) with a 20-fold selectivity at the M3/M1 receptors, was identified. Quantitative structure-activity relationships (QSAR) showed that the size of the ester was the main structural feature determining both binding affinity for the M3/M1 receptors and inhibitory activity on guinea pig bronchi. Esters with substituted acyl side chains (fewer hyperconjugable H atoms at the alpha-carbon) are generally associated with better activity and affinity.

Eurycoma longifolia increases sexual motivation in sexually naive male rats.

The aim of this study is to provide evidence on the aphrodisiac property of Eurycoma longifolia Jack. An electric grid was used as an obstruction in the electrical copulation cage in order to determine how much an aversive stimulus the sexually naive male rat for both the treated with E. longifolia Jack and control groups were willing to overcome to reach the estrous receptive female in the goal cage. The intensity of the grid current was maintained at 0.12 mA and this was the intensity in which the male rats in the control group failed to crossover to reach the goal cage. Results showed that E. longifolia Jack continued to enhance and also maintain a high level of both the total number of successful crossovers, mountings, intromissions and ejaculations during the 9-12th week observation period. In conclusion, these results further enhanced and strengthened the aphrodisiac property of E. longifolia Jack.