RESUMO
Strong blue fluorescence at 487 nm corresponding to the (1)G(4) --> (3)H(6) transition was generated from Tm(3+)-doped lutetium aluminum garnet on excitation with a 618-nm dye laser as a result of a photon-avalanche upconversion mechanism.
RESUMO
The liver has been judged relatively resistant to ischemia, but prolonged inflow occlusion at normothermic conditions can produce evidence of reversible or irreversible hepatocellular damage. Cytoprotective agents have been used both experimentally and clinically to afford extended viability of hepatocytes under reduced perfusion. One agent, prostaglandin E1, has been described clinically as effective in sustaining liver function under ischemic conditions. We have sought to verify this observation in an experimental model using prolonged normothermic inflow occlusion. Twenty miniature pigs were anesthetized and subjected to subtotal normothermic hepatic inflow occlusion (portal vein, hepatic artery, choledochal vessels) to allow for sufficient splanchnic decompression. Half of the animals received pretreatment with prostaglandin E1 (alprostadil) 500 micrograms intravenously. Inflow occlusion was maintained for 2 hours followed by reperfusion and killing 24 hours later. As a measure of functional preservation, the tissue adenine nucleotides adenosine monophosphate, diphosphate, and triphosphate (AMP, ADP, ATP) were measured in ischemic liver by freeze-clamping and high-performance liquid chromatography during occlusion and after reperfusion. Cytosolic enzyme determinations (aspartate transaminase, alanine transaminase, lactate dehydrogenase) were also made before occlusion and after reperfusion. As a possible indicator of cellular injury, blood ionized Ca++ was measured before inflow occlusion and after reperfusion. Although no difference was found in levels of AMP and ADP between prostaglandin E1 and control animals, ATP levels rose significantly higher during recovery in prostaglandin E1 animals at 60 minutes and 24 hours after reperfusion (13.97 +/- 1.29 and 13.60 +/- 0.91 mumoles/gm dry weight prostaglandin E1 vs. 9.25 +/- 0.97 and 9.80 +/- 0.85 mumoles/gm dry weight co control, P < .01).(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Nucleotídeos de Adenina/metabolismo , Alanina Transaminase/metabolismo , Alprostadil/farmacologia , Aspartato Aminotransferases/metabolismo , Isquemia/metabolismo , L-Lactato Desidrogenase/metabolismo , Fígado/irrigação sanguínea , Fígado/metabolismo , Animais , Feminino , Injeções Intravenosas , Isquemia/prevenção & controle , Fígado/patologia , SuínosRESUMO
OBJECTIVE: To determine the median response time to therapy with vancomycin alone or with vancomycin plus rifampin in patients with methicillin-resistant Staphylococcus aureus (MRSA) endocarditis. DESIGN: Cohort analysis of a randomized study. SETTING: University medical center. PATIENTS: Forty-two consecutive patients with MRSA endocarditis were randomly assigned to receive either vancomycin (group I) or vancomycin plus rifampin (group II) for 28 days. MEASUREMENTS: Clinical signs and symptoms were recorded, and blood cultures were obtained daily to determine the duration of bacteremia. MAIN RESULTS: The median duration of bacteremia was 9 days (7 days for group I and 9 days for group II). The median duration of fever for all patients and for each treatment group was 7 days. Six patients failed therapy, including three patients who died 5, 6, and 9 days after therapy was started, respectively. The other three patients who failed therapy required valve surgery on days 2, 22, and 27, respectively. Although patients had sustained bacteremia, no unusual complications were seen in either treatment group, and most patients responded to continued antibiotic therapy. CONCLUSIONS: Slow clinical response is common among patients with MRSA endocarditis who are treated with vancomycin or vancomycin plus rifampin. Nevertheless, few complications appear to be related solely to this sustained bacteremia.
Assuntos
Endocardite Bacteriana/tratamento farmacológico , Rifampina/uso terapêutico , Infecções Estafilocócicas/tratamento farmacológico , Vancomicina/uso terapêutico , Adulto , Bacteriemia/tratamento farmacológico , Quimioterapia Combinada/uso terapêutico , Endocardite Bacteriana/microbiologia , Feminino , Febre/tratamento farmacológico , Febre/microbiologia , Gentamicinas/uso terapêutico , Humanos , Masculino , Resistência a Meticilina , Pessoa de Meia-Idade , Estudos Prospectivos , Estatística como Assunto , Fatores de TempoRESUMO
STUDY OBJECTIVE: The aim was to determine whether the potent iron chelator desferrioxamine, previously shown by our laboratory and others to cause acute limitation of infarct size, would provide sustained protection against myocardial ischaemia-reperfusion injury. DESIGN: Anaesthetised dogs underwent a 2 h transient coronary artery occlusion. After surgical preparation, each dog was randomised into one of two groups receiving either desferrioxamine or equivalent infusion of saline. Infusion of desferrioxamine was initiated 30 min before occlusion at an initial dose of 10 mg.kg-1 for the first hour of the protocol, followed by a maintenance dose of 1.5 mg.kg-1.h-1 throughout the remainder of the ischaemic period and the initial 4 h of reperfusion. The animals were reanaesthetised the following day and killed 20-24 h following reperfusion. Variables measured included heart rate and arterial pressures; regional myocardial blood flow; urinary iron content; and infarct size. In addition, % wall thickening in the ischaemic-reperfused myocardium was assessed by echocardiography in a limited number of animals. EXPERIMENTAL MATERIAL: 12 mongrel dogs weighing 22(SEM 4) kg were used, n = 6 in each group. MEASUREMENTS AND MAIN RESULTS: Both groups were equally and severely ischaemic during coronary artery occlusion. Desferrioxamine caused a modest and transient reduction in mean arterial pressure during the ischaemic episode, but had no effect on heart rate or myocardial blood flow. As expected, urinary iron content was significantly higher in treated animals v controls, at 465(SEM 107) v 55(23) micrograms, p less than 0.01, indicating that desferrioxamine effectively chelated free iron. However, it failed to exert a significant cardioprotective effect: infarct size did not differ between control and treated groups [54(4)% v 58(5)% of the myocardium at risk], and wall thickening was similar throughout occlusion and reperfusion in control and desferrioxamine treated animals. CONCLUSIONS: Despite substantial reduction in infarct size previously observed at 4 h following reflow with the same dose and regimen of desferrioxamine treatment, results of the present study indicate that desferrioxamine did not provide sustained protection against myocardial ischaemia-reperfusion injury. We therefore conclude that desferrioxamine delays--but does not prevent--myocyte necrosis in this canine model.
Assuntos
Desferroxamina/uso terapêutico , Traumatismo por Reperfusão Miocárdica/prevenção & controle , Animais , Pressão Sanguínea/efeitos dos fármacos , Cães , Ecocardiografia , Feminino , Ferro/urina , Masculino , Infarto do Miocárdio/patologia , Miocárdio/patologia , Distribuição Aleatória , Fatores de TempoRESUMO
For the successful development of a high-affinity fluorophore-estradiol conjugate, the fluorophore must be attached to the estradiol molecule at a position that interferes least with its binding to the receptor. We have concentrated on 17 alpha substituents as models for fluorophore attachment, based on literature precedent and on our earlier work with small 17 alpha side chains. In this report, we describe syntheses and estrogen receptor binding affinities of 19 analogs of estradiol substituted in the 17 alpha position with larger side chains (of six to 11 carbons), some of which may be synthetically modified to link a fluorophore. These analogs were synthesized either by nucleophilic cleavage of estrone-17 beta-oxirane 3-benzyl ether and subsequent debenzylation (4 to 18), by cross-coupling of alkynes (21 to 24), by alkylation of 17 alpha-ethynylestradiol 3,17-bis(tetrahydropyranyl ether) and subsequent acidic hydrolysis (25 to 28), or by reacting estrone either with appropriate aryl/alkynyllithium reagents (29, 30, and 32) or with benzylmagnesium bromide (31). Relative binding affinities of these newly synthesized analogs were determined for estrogen receptor (rat uterus) using a standard competition assay. The results suggest that analogs with reduced mobility and/or more polarizable electron density in the side chain generally bind more strongly to the receptor. The relative affinities of several selected compounds were also determined in the presence of 4% dimethylformamide; some compounds bearing larger, nonpolar 17 alpha substituents showed dramatically improved affinities, while affinities for compounds with shorter nonpolar side chains remained largely unchanged. These binding affinity results should be useful in designing new high-affinity fluorescent ligands for the estrogen receptor.
Assuntos
Receptores de Estradiol , Fluorescência , Ligantes , Espectroscopia de Ressonância MagnéticaRESUMO
In order to develop stable, high-affinity fluorescein-estradiol conjugates, the fluorescein moiety must be leashed to the estradiol molecule at a point which interferes least with estradiol's binding to the receptor. Because of the high affinity of 17 alpha-substituted estradiol (e.g. ethynyl estradiol), we investigated a series of 17 alpha-substituted estradiol compounds to determine the optimal properties of a leash at this position. Twelve estradiol derivatives bearing a three-carbon 17 alpha side chain with or without a terminal functional group and with varying degrees of unsaturation were synthesized. Initial comparison of the receptor binding affinities of some of these derivatives suggested that three factors might reduce affinity: internal hydrogen bonding of the 17 beta-hydroxyl proton with an oxygen atom of the 17 alpha side chain; hydrophilicity of the ligand; or steric interference of the side chain with receptor binding. Further comparisons were designed to evaluate the relative contribution of these factors. The results suggest that the relative affinities of these 17 alpha-substituted estradiol derivatives are influenced primarily by the steric interference of the side chains and also by their hydrophilicity. Internal hydrogen bonding involving the 17 beta-hydroxyl proton does not seem to have a profound effect.