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The feeding ecology of two reef fishes associated with artificial reefs in the northwest Gulf of Mexico (GoM) was examined using gut contents and natural stable isotopes. Reefs were divided into three regions (east, central, west) across an east to west gradient of increasing reef complexity and salinity. Gray triggerfish (Balistes capriscus) primarily consumed reef-associated prey (xanthid crabs, bivalves, barnacles) and pelagic gastropods, while red snapper (Lutjanus campechanus) diets were mainly comprised of non-reef prey (stomatopods, fishes, portunid crabs). Natural stable isotopes of carbon (δ13C), nitrogen (δ15N), and sulfur (δ34S) were measured in consumer muscle tissue as well as potential primary producers. Gray triggerfish occupied a lower trophic position than red snapper, with lower δ13C and δ15N values across all size classes and regions, and generally higher δ34S values. Red snapper had a smaller range of stable isotope values and corrected standard ellipse areas across all size classes and regions, indicating a smaller isotopic niche. Contribution estimates of particulate organic matter (26 to 54%) and benthic microalgae (BMA, 47 to 74%) for both species were similar, with BMA contributions greater across all three size classes (juveniles, sub-adults, adults) of red snapper and all but the juvenile size class for gray triggerfish. Species gut contents and stable isotopes differed by region, with fishes consuming more crabs in the east region and more gastropods in the central and west regions. δ13C and δ15N values generally decreased from east to west, while δ34S increased across this gradient. Results highlight species-specific feeding differences associated with artificial reefs, where gray triggerfish may be more dependent on the reef structure for foraging opportunities. In addition, results offer further information on the integral role of BMA in primary production at nearshore artificial reefs., Competing Interests: The authors have declared that no competing interests exist.

Heat shock (HS) results in the expression of heat shock proteins (hsp) and confers tolerance against subsequent ischemic injury. We examined the extent of myocardial protection in vivo, and determined the level of hsp expression induced by HS as a function of time. Anesthetized rats were subjected to HS by raising core temperature to 42 degrees C for 15 min and they were then allowed to recover from 2 to 30 h (n = 8-11 for each time point). At the appropriate time, animals were subjected to 30 min of ischemia via ligation of the LAD, followed by 90 min of reperfusion. Infarct size was determined by tetrazolium staining and hsp expression was assessed by Western blots. Following ischemia/reperfusion, the infarct sizes (% risk area) were 51.3 +/- 3.7, 41.0 +/- 7.7, 48.0 +/- 6.9 after 2, 4 and 12 h of HS, which were not significantly different from 39.2 +/- 2.75 in non heat-shocked animals (P > 0.05). In contrast, the infarct size was reduced significantly to 11.0 +/- 3.1% in 24 h HS group (P < 0.01 v non-heat-shocked control, 2, 4 and 12 h HS groups), but increased back to 40.0 +/- 3.2% (P < 0.01) by 30 h after HS. No major significant differences in the mean arterial blood pressure, heart rate or rate pressure product was observed between different groups. The synthesis of 72- and 27-kD hsp in HS groups was rapid, reaching > 80% of maximum within 4 h of initial insult and peaked by 12 h, whereas the protective effect of HS was absent at these time points. We conclude that ischemic tolerance afforded by HS cannot be solely explained on the basis of hsp expression, and may be dependent on factors such as post-translational modifications, translocation of hsps or some other as yet unidentified factors.

Heat stress (HS) and the subsequent expression of heat shock proteins has been shown to enhance post-ischemic functional recovery and reduce infarct size. The purpose of these experiments was to determine if HS pre-treatment preserves sarcoplasmic reticulum (SR) function, a cellular organelle that plays an important role in myocardial contractility. Anesthetized rats were heat stressed for 15 min by raising temperature to 42 degrees C. Twenty-four hours later the hearts were perfused by Langendorff's method and subjected to either 20 or 35 min of global ischemia, with a subset of hearts then being subjected to 10 or 20 min of reperfusion, respectively. SR function was assessed by oxalate-supported Ca2+ uptake rate in cell free preparations in the presence and absence of ruthenium red, a selective SR calcium release channel blocker Ca2+ uptake decreased significantly from 25.6 +/- 3.4 to 13.4 +/- 1.9 and 11.3 +/- 2.3 nmol/min/mg protein (mean +/- S.E.), following 20 and 35 min of ischemia, respectively. A similar trend was observed following reperfusion as well. No significant difference in Ca2+ uptake was observed between HS v control hearts. Similarly, in samples where the Ca2+ release channel was blocked with ruthenium red, decreased Ca2+ uptake rates were noted after both ischemia and reperfusion, with no significant differences seen between HS and non-HS hearts. There was significant improvement it developed pressure. +dP/dt and -dP/dt, with reduced creatine kinase release in HS v non-HS hearts. Western blot analysis demonstrated increased synthesis of 27- and 70-kDa heat shock proteins in HS but not in control animals. It is concluded that HS improves functional recovery and induces expression of 27- and 70-kDa heat shock proteins without preservation of SR function in the globally ischemic rat heart.

The purpose of this study was to evaluate the protective effect of a new endotoxin analogue, monophosphoryl lipid A (MLA) in a rabbit model of myocardial ischemia/reperfusion and to show if this protection was mediated via synthesis of 70 kDa heat shock protein (HSP 70). Three groups of New Zealand White rabbits underwent 30 min coronary occlusion, followed by 4 hours reperfusion. First group of rabbits (n = 6) were treated with 0.35 ml vehicle (40% propylene glycol, 10% ethanol in water). The second and third group of rabbits (n = 6-8) were treated with MLA (35 micrograms/kg, i.v.) 12 and 24 hours prior to ischemia and reperfusion. MLA treatment either 12 or 24 h prior to ischemia/reperfusion demonstrated significantly reduced infarct size (12.5 +/- 1.7 and 14.7 +/- 2.1% for 12 and 24 h) when compared with vehicle control (40.4 +/- 8.6%, mean +/- S.E.M, p < 0.05). No significant differences in the infarct size was observed between the 12 and 24 h MLA treated groups. The area at risk was not significantly different between the three groups. Baseline values of heart rate, systolic and diastolic blood pressure were not significantly different between the control and MLA treated groups. However, the systolic as well as diastolic blood pressure during reperfusion were significantly lower in rabbits treated with MLA. Western blot analysis of the protein extracts of the hearts (n = 2/group) demonstrated no increase in the expression of the inducible form of HSP 70 following treatment with MLA. We conclude that MLA has significant anti-infarct effect in rabbit which is not mediated by the cardioprotective protein HSP 70. The anti-infarct effect of this drug is superior to the reported protective effects of delayed ischemic or heat stress preconditioning. We hypothesize that the pharmacologic preconditioning afforded by MLA is accomplished via a unique pathway that bypasses the usual intracellular signaling pathways which lead to the myocardial protection with the expression of heat shock proteins.

Milrinone (Inocor-Sanofi-Winthrop) represents a second generation phosphodiesterase inhibitor currently approved for intravenous administration in the treatment of decompensated congestive heart failure. By inhibiting Type III phosphodiesterase, milrinone increases intracellular cyclic adenosine monophosphate. This results in a positive inotropic effect on the heart and vasodilatation in the periphery. The hemodynamic consequences of this action produce left ventricular afterload reduction, with an increase in cardiac output and a reduction in total peripheral resistance. Unlike the sympathomimetic amines, milrinone produces no tolerance and possesses the distinct advantage of directly decreasing pulmonary vascular resistance. Short-term intermittent infusion by peripheral administration, continuous infusion, long-term therapy, and intermittent outpatient therapy was demonstrated to be safe, efficacious, and cost effective. It is hypothesized that intravenous milrinone, by producing biventricular afterload reduction, offers an efficacious, cost-effective tool for the treatment of decompensated heart failure.

The purpose of this study was to evaluate the protective effect of a new endotoxin analogue, monophosphoryl lipid A (MLA) in a rabbit model of myocardial ischemia/reperfusion and to show if this protection was mediated via synthesis of 70 kDa heat shock protein (HSP 70). Three groups of New Zealand White rabbits underwent 30 min coronary occlusion, followed by 4 hours reperfusion. First group of rabbits (n = 6) were treated with 0.35 ml vehicle (40 % propylene glycol, 10 % ethanol in water). The second and third group of rabbits (n = 6-8) were treated with MLA (35 micrograms/kg, i.v.) 12 and 24 hours prior to ischemia and reperfusion. MLA treatment either 12 or 24 h prior to ischemia/reperfusion demonstrated significantly reduced infarct size (12.5 +/- 1.7 and 14.7 +/- 2.1% for 12 and 24 h) when compared with vehicle control (40.4 +/- 8.6%, mean +/- S.E.M, p < 0.05). No significant differences in the infarct size was observed between the 12 and 24 h MLA treated groups. The area at risk was not significantly different between the three groups. Baseline values of heart rate, systolic and diastolic blood pressure were not significantly different between the control and MLA treated groups. However, the systolic as well as diastolic blood pressure during reperfusion were significantly lower in rabbits treated with MLA. Western blot analysis of the protein extracts of the hearts (n = 2/group) demonstrated no increase in the expression of the inducible form of HSP 70 following treatment with MLA. We conclude that MLA has significant anti-infarct effect in rabbit which is not mediated by the cardioprotective protein HSP 70. The anti-infarct effect of this drug is superior to the reported protective effects of delayed ischemic or heat stress preconditioning. We hypothesize that the pharmacologic preconditioning afforded by MLA is accomplished via a unique pathway that bypasses the usual intracellular signaling pathways which lead to the myocardial protection with the expression of heat shock proteins.

Congestive heart failure (CHF) is a common clinical entity with diverse causes. Patients may present with acute decompensation or have a more indolent course, with diminishing exercise tolerance and increasing dyspnea. The management of this clinical entity traditionally has focused on restricting fluid intake, decreasing dietary sodium, decreasing afterload with vasodilatory agents, and supporting the failing myocardium with agents which produce a positive inotropic response. In the acutely decompensated patient, short-term therapy with positive inotropics is clearly beneficial. The role of long-term inotropic therapy for chronic CHF remains less clear. A number of clinical trials have recently evaluated the effects of long-term therapy on morbidity and mortality, with disappointing results. For a number of the newer, nonglycocide oral positive inotropics, at doses of drug which produce measurable hemodynamic improvement, increased mortality in treatment groups has been unacceptably high. Ironically, patients with worse left ventricular dysfunction show the most clinical improvement, but have the highest increased mortality. However, as with digoxin, there is some evidence that employing lower doses of drug that do not produce measurable improvement in hemodynamic parameters may result in both improved clinical state and decreased mortality. This review discusses the role of both oral and intravenous inotropic agents, discusses the difficulty in translating short-term hemodynamic improvement into long-term clinical benefit, and presents a rationale for the use of lower-dose inotropic therapy to improve long-term clinical outcome.

Study Objectives: To evaluate continuous, semiquantitative end-tidal carbon dioxide (ETCO2) monitoring in the prehospital and emergency department setting for confirming proper endotracheal tube placement and assessing prognosis and blood flow during CPR., Type of Participants: Adult patients were included if an endotracheal tube was inserted by prehospital care providers or emergency physicians for cardiac arrest, respiratory arrest, respiratory insufficiency, or airway protection., Design and Interventions: A small, portable, colorimetric ETCO2 detector was attached to the endotracheal tube immediately after each attempted endotracheal tube insertion. The color of the detector membrane was noted at the seventh breath following intubation. The color also was noted and recorded if there was return of spontaneous circulation (defined as a palpable pulse) immediately prior to and following conversion from manual to mechanical CPR. Survival to hospital admission was used as an end point to assess the prognostic value of the initial ETCO2 reading., Main Results: A total of 227 patients (144 with cardiopulmonary arrest) were studied. In the 83 patients intubated but not in cardiopulmonary arrest, a reading on the ETCO2 detector signifying more than 0.5% ETCO2 was 100% sensitive and 93% specific in detecting proper endotracheal tube placement (100% specific with the endotracheal tube cuff inflated). In cardiac arrest patients, a longer period of estimated arrest appeared to be associated with a lower ETCO2 detector reading. A reading signifying more than 0.5% ETCO2 was 69% sensitive and 100% specific in detecting proper endotracheal tube placement. After proper endotracheal tube placement, all cardiac arrest patients who survived to hospital admission had an initial ETCO2 measurement signifying more than 0.5% ETCO2. Return of spontaneous circulation was usually accompanied by an improved ETCO2 value. Mechanical CPR always produced an ETCO2 value that was as high or higher than that produced by manual CPR., Conclusion: The colorimetric ETCO2 device is highly accurate for confirming endotracheal tube position in nonarrest patients., Conclusion: The colorimetric ETCO2 device is highly accurate for confirming endotracheal tube position in nonarrest patients. In cardiac arrest patients, a reading signifying more than 0.5% ETCO2 confirms correct endotracheal tube placement, while a value signifying less than 0.5% ETCO2 during resuscitation suggests that something is wrong (eg. esophageal intubation, inadequate circulatory flow, prolonged down-time interval, hypothermia, or significant ventilation/perfusion mismatch).

Certain properties of the bleomycin analogs deglycobleomycin A2 and decarbamoylbleomycin A2 have been characterized. In common with bleomycin A2, both deglycobleomycin A2 and decarbamoylbleomycin A2 were found to mediate DNA degradation in the presence of Fe(II) + O2. Both analogs were found to have essentially the same sequence selectivity for DNA strand scission as bleomycin A2 when a 5'-[23P]-end-labeled linear duplex DNA derived from SV40 DNA was employed as a substrate. Product analysis for the three analogs was carried out by assay for malondialdehyde (precursors) after digestion of calf thymus DNA, and also by hplc analysis of the digestion products formed from the dodecanucleotide d(CGCTTTAAAGCG). All three Fe(II) X bleomycin A2 analogs produced the same products, albeit not in the same relative amounts.

DNA strand scission by bleomycin in the presence of Cu and Fe was further characterized. It was found that DNA degradation occurred readily upon admixture of Cu(I) or Cu(II) + dithiothreitol + bleomycin, but only where the order of addition precluded initial formation of Cu(II)--bleomycin or where sufficient time was permitted for reduction of the formed Cu(II)--bleomycin to Cu(I)--bleomycin. DNA strand scission mediated by Cu + dithiothreitol + bleomycin was inhibited by the copper-selective agent bathocuproine when the experiment was carried out under conditions consistent with Cu chelation by bathocuproine on the time scale of the experiment. Remarkably, it was found that the extent of DNA degradation obtained with bleomycin in the presence of Fe and Cu was greater than that obtained with either metal ion alone. A comparison of the sequence selectivity of bleomycin in the presence of Cu and Fe using 32P-end-labeled DNA duplexes as substrates revealed significant differences in sites of DNA cleavage and in the extent of cleavage at sites shared in common. For deglycoblemycin and decarbamoylbleomycin, whose metal ligation is believed to differ from that of bleomycin itself, it was found that the relative extents of DNA cleavage in the presence of Cu were not in the same order as those obtained in the presence of Fe. The bleomycin-mediated oxygenation products derived from cis-stilbene were found to differ in type and amount in the presence of added Cu vs. added Fe. Interestingly, while product formation from cis-stilbene was decreased when excess Fe was added to a reaction mixture containing 1:1 Fe(III) and bleomycin, the extent of product formation was enhanced almost 4-fold in reactions that contained 5:1, as compared to 1:1, Cu and bleomycin. The results of these experiments are entirely consistent with the work of Sugiura [Sugiura, Y. (1979) Biochem. Biophys. Res. Commun. 90, 375-383], who first demonstrated the generation of reactive oxygen species upon admixture of O2 and Cu(I)--bleomycin.

The matrix (M) protein of vesicular stomatitis virus (VSV) has a major antigenic determinant (epitope 1) that maps to a region extending from amino acids 19 through 43 and transcription-inhibition activity that maps to the first 43 N-terminal amino acids (J.R. Ogden, R. Pal, and R. R. Wagner, J. Virol. 58:860-868, 1986). The M protein of temperature-sensitive mutant tsO23(III) is devoid of epitope 1 and transcription-inhibition activity and substitutes glutamic acid for glycine at amino acid 21 as well as having amino acid substitutions at positions 111 and 227 (K. Morita, R. Vanderoef, and J. Lenard, J. Virol. 61:256-263, 1987). We undertook to map more precisely epitope 1 and the transcription-inhibition region of VSV M protein by means of synthetic oligopeptides generated by an automated solid-phase protein synthesizer. A pentadecapeptide designated PI(wt, Gly21), corresponding to amino acids 17 to 31 of wild-type (wt) M protein, strongly bound monoclonal antibody MAb2 (directed to epitope 1); however, an analogous pentadecapeptide with glutamic acid substituted for glycine at position 21, designated PII(tsO23, Glu21), completely failed to recognize MAb2. Polyclonal antibody raised in rabbits immunized with PI(wt, Gly21) reacted strongly with wt M protein, the homologous pentadecapeptide, and, to a lesser extent, PII(tsO23, Glu21). Anti-PII(tsO23, Glu21) failed to recognize PI(wt, Gly21) or wt M protein. Anti-PI(wt, Gly21) competed efficiently for binding of MAb2 to wt M protein and was as effective as MAb2 in reversing inhibition of VSV transcription by wt M protein. Neither PI(wt, Gly21) nor PII(tsO23, Glu21) exhibited any ability to inhibit VSV transcription. However, a lysine-rich oligopeptide, PII(Met1-Leu20), corresponding to the first 20 N-terminal amino acids of wt M protein, and polylysine itself did inhibit VSV transcription, albeit much less efficiently than native wt M protein. Monospecific polyclonal antibody directed to the 20-mer oligopeptide PIII(Met1-Leu20) reversed transcription inhibition by M protein in a dose-dependent manner almost identical to that of anti-PI(wt, Gly21) and epitope 1-specific MAb2. Examination by circular dichroism spectropolarimetry revealed significant differences in the conformation of the two pentadecapeptides attributable to the Gly in equilibrium Glu amino acid substitution at position 21.