Long-lasting antinociceptive effect of DAMGO chloromethyl ketone in rats

Previously, the opioid peptide Tyr-D-Ala-Gly-(NMe)Phe-CH2Cl (DAMCK) has been shown to bind irreversibly to mu opioid receptors in vitro. In the present work, the antinociceptive effect of DAMCK has been evaluated. Rats treated systemically with DAMCK (1-100 pg/kg) displayed a dose-dependent increase in tail-flick analgesia that peaked by 15 min, then stayed about the same until 60 min, followed by some decrease over time. Higher doses of DAMCK (10 ng/kg-100 microg/kg) produced a near-maximal antinociceptive effect that remained stable for 4 h. Significant antinociception was still detected 8 h, but not 24 h postinjection. In comparison, the parent peptide DAMGO (Tyr-D-Ala-Gly-(NMe)Phe-Gly-ol) reached maximal effect by about 30 min, followed by a rapid cessation of its antinociceptive response. Naloxone administered before DAMCK antagonized the antinociceptive response of DAMCK, indicating that it was mediated via opioid receptors. Naloxone administered 45 min after DAMCK attenuated the tail-flick response to some extent, but a substantial part (40-60% depending on the peptide concentration and evaluation time) remained unaffected. Central administration of DAMCK also elicited time- and concentration-dependent, profound, opioid receptor mediated, apparently irreversible antinociception.     

Selective inhibition of neuronal nitric oxide synthase fails to alter the resting tension and the relaxant effect of bradykinin in isolated rat middle cerebral arteries

The role of the neuronal isoform of the nitric oxide (NO) synthase (nNOS) in the regulation of the cerebrovascular tone was studied in vitro. Selective inhibition of nNOS by 7-nitro indazole monosodium salt (7-NINA) failed to alter the resting tension and the relaxant effect of bradykinin in isolated rat middle cerebral arteries. These results indicate that 1./ 7-NINA is selective for nNOS and 2./ cerebrovascular nNOS is involved neither in the resting NO production nor in the mediation of the relaxant effect of bradykinin. Therefore, nNOS-derived NO that contributes to the maintenance of the resting cerebral blood flow in vivo appears to be released from neurons and/or glial cells.     

Cold pressor test and retinal capillary perfusion in vasospastic subjects with and without capsular glaucoma (a preliminary study)

PURPOSE: Using the cold pressor test the authors investigated the change in retinal and neuroretinal capillary perfusion in vasospastic patients suffering from capsular glaucoma (CG) and in vasospastic control subjects. METHODS: Changes in retinal and optic nerve head capillary perfusion induced by the cold pressor test (one hand immersed in 4 degrees C water for 30 seconds, then in 30 degrees C water for 2 minutes) was measured using the Heidelberg Retina Flowmeter in 4 patients with CG and in 5 healthy control subjects. Previously all subjects showed a reduction of cutaneous capillary flow higher than 70% in the cold pressor test (vasospastic reaction). One eye per subject was investigated. Two images were obtained for each phase (baseline, cold phase and warm phase), and the better quality image from each phase was selected for the measurements. One location on the temporal neuroretinal rim and one location on the temporal retina outside the peripapillary area were selected for the HRF measurements. RESULTS: In the CG group neuroretinal rim "Volume" decreased by 26.05%, "Flow" by 25.82% and "Velocity" by 23.91% (p<0.05), retinal "Volume" decreased by 12.30% (p=0.051), and retinal "Flow" by 22.36% (p=0.01) in the cold phase. All these parameters returned to the corresponding baseline values in the warm phase. In the control group a significant decrease was observed in retinal "Volume" (15.96%), "Flow" (17.81%), and "Velocity" (16.11%) in the cold phase (p<0.05), which diminished in the warm phase but remained still significant for "Flow" and "Velocity". CONCLUSION: Cutaneous cold provocation can induce an immediate decrease in retinal and optic nerve head capillary perfusion at least in a part of the vasospastic subjects with or without capsular glaucoma. This decrease diminishes or disappears quickly when the hand is immersed in warm water. To evaluate the potential role of cold-induced retinal and optic nerve head vasoconstriction in the pathogenesis of capsular glaucoma further investigations are necessary since this reaction was also present in the vasospastic control subjects.     

Application of an anti-HQIgY antibody for the measurement of IgY concentrations of hen's and quail's serum and yolk

The development of a sensitive ELISA for the measurement of quail IgY (QIgY) was the main purpose of our study. The suitable antibody (AB) was prepared in rabbits. Both quail IgY (QIgY) and hen IgY (HIgY) were precipitated by this developed AB. For this reason it was marked as anti-hen-quail-IgY (a-HQIgY). The purified AB was conjugated with horseradish peroxidase (aHQIgY-HRP) and a sensitive direct ELISA was developed, based on this labeled AB. The prepared aHQIgY AB which was used in this developed ELISA method was suitable for the measurement of total and specific IgY concentration in domestic hen (Gallus domesticus) and Japanese quail (Coturnix coturnix japonica) either. As a result of our experiments it is very likely that there are identical sequences of IgYs of both species. This part of IgY has good antigen character at the same time. Probably, this phenomenon has occurrence in other Galliform species, too. Further investigations will be carried out in this field.     

Hypoxia/reoxygenation and vitamin C intake influence NO synthesis and antioxidant defenses of neutrophils

Oxidative stress induced by hypoxia/reoxygenation mediates the pathophysiological consequence of ischemia/reperfusion and human diseases. Diving apnea could be a good model of oxidative stress induced by hypoxia/reoxygenation. We studied the influence of vitamin C diet supplementation on the response of neutrophil antioxidant defenses, NO production, and redox status to diving apnea. Seven professional apnea divers participated in a double-blind cross study. Divers were assigned to either vitamin C-supplemented (1 g/d for a week) or placebo groups. Blood samples were taken under basal conditions, immediately after diving apnea for 4 h and after 1 h of recovery. Plasma vitamin C increased only in the supplemented group after diving and was maintained high in recovery. Diving apnea decreased neutrophil GSH/GSSG ratio in both groups, but maintained protein carbonyl derivates. Neutrophil catalase activity and levels and glutathione peroxidase activity were lower in the supplemented group than in the placebo group after diving. iNOS and nitrite levels decreased only in the supplemented group after diving and recovery. Diving apnea induced oxidative stress and initiated neutrophil reactions that resemble the acute-phase immune response with increased myeloperoxidase activity in neutrophils. Diet supplementation with vitamin C reduced neutrophil iNOS levels and NO production.     

Left ventricular pressure-volume relationship in a rat model of advanced aging-associated heart failure

Aging is associated with profound changes in the structure and function of the heart. A fundamental understanding of these processes, using relevant animal models, is required for effective prevention and treatment of cardiovascular disease in the elderly. Here, we studied cardiac performance in 4- to 5-mo-old (young) and 24- to 26-mo-old (old) Fischer 344 male rats using the Millar pressure-volume (P-V) conductance catheter system. We evaluated systolic and diastolic function in vivo at different preloads, including preload recruitable stroke work (PRSW), maximal slope of the systolic pressure increment (+dP/dt), and its relation to end-diastolic volume (+dP/dt-EDV) as well as the time constant of left ventricular pressure decay, as an index of relaxation. The slope of the end-diastolic P-V relation (EDPVR), an index of left ventricular stiffness, was also calculated. Aging was associated with decrease in left ventricular systolic pressure, +dP/dt, maximal slope of the diastolic pressure decrement, +dP/dt-EDV, PRSW, ejection fraction, stroke volume, cardiac and stroke work indexes, and efficiency. In contrast, total peripheral resistance, left ventricular end-diastolic volume, left ventricular end-diastolic pressure, and EDPVR were greater in aging than in young animals. Taken together, these data suggest that advanced aging is characterized by decreased systolic performance accompanied by delayed relaxation and increased diastolic stiffness of the heart in male Fischer 344 rats. P-V analysis is a sensitive method to determine cardiac function in rats.     

Feeding habits of the fishes Lutjanus peru and Lutjanus guttatus (Pisces: Lutjanidae) of Guerrero, México

Diet composition of the Pacific snapper (Lutjanus peru) 130-684 mm fork length (FL) and the spotted snapper (Lugjanus guttatus) 120-550 mm FL, was analyzed. Monthly samples were obtained from commercial landings in three regions off the coast of Guerrero, Mexico. Percentage by number (%N), percentage by weight (%W), and percentage of occurrence (%O) were calculated for each prey and summarized as the index of relative importance. Both species are polyphagous predators feeding on a variety of prey: Sixty-eight prey items, mainly fish (%W = 50.9), crustaceans (%W = 35.6), and mollusks (%W = 7.2), were found in the stomach contents of L. peru, while 88 components were identified in the diet of L. guttatus, the most important prey being fish (%W = 50.8) and crustaceans (%W = 43.4). Diet overlap between species is not significant (p<0.05), indicating that competition for food is unlikely. Based on published values of the relative importance index, the diets of these Lutjanidae were analyzed considering different regions of the Pacific coast of Mexico and Costa Rica, and similarities among sites and species were discussed. The cluster analysis showed that similarities among species inhabiting in the same area are more important than within organisms of the same species living in different zones. Results suggest that prey availability rather than food selectivity, conditions the feeding behavior of these fish species.