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Biomedical subjects

R Ray

Publications and source records attributed to R Ray.

At least 217 records · Page 12Linked to original sources

Photoaffinity labeling of the rat plasma vitamin D binding protein with [26,27-3H]-25-hydroxyvitamin D3 3 beta-[N-(4-azido-2-nitrophenyl)glycinate].

It is well recognized that the vitamin D binding protein (DBP) is important for the transport of vitamin D, 25-hydroxyvitamin D (25-OH-D), and its metabolites. In an attempt to better understand the molecular-binding properties of this ubiquitous protein, we designed and synthesized a photoaffinity analogue of 25-OH-D3 and its radiolabeled counterpart. This analogue, 25-hydroxyvitamin D3 3 beta-[N-(4-azido-2-nitrophenyl)glycinate] (25-OH-D3-ANG), was recognized by the rat DBP and was about 10 times less active than 25-OH-D3 in terms of binding. Incubation of [3H]25-OH-D3 or [3H]25-OH-D3-ANG with rat DBP revealed that both compounds were specifically bound to a protein with a sedimentation coefficient of 4.1 S. Each was displaced with a 500-fold excess of 25-OH-D3. When [3H]25-OH-D3-ANG was exposed to UV radiation in the presence of rat DBP followed by the addition of a 500-fold excess of 25-OH-D3, there was no displacement of tritium from the 4.1S peak. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis autoradiographic analysis of [3H]25-OH-D3-ANG exposed to UV radiation in the presence of rat DBP followed by the addition of a 500-fold excess of 25-OH-D3 revealed one major band with a molecular weight of 52 000. These data provide strong evidence that [3H]25-OH-D3-ANG was covalently linked to the rat DBP. This photoaffinity probe should provide a valuable tool for the analysis of the binding site on this transport protein.

Affinity Labels↗

Inhibition of acetylcholinesterase from electric eel by (-)-and (+)-physostigmine and related compounds.

Unnatural (+)-physostigmine (2) inhibited acetylcholinesterase (AChE) from electric eel considerably less than natural (-)-physostigmine (1), but 2 may because of its possible lower toxicity still be an interesting anticholinesterase agent. (-)-Eseroline (3), a major metabolite of (-)-physostigmine (1) and a potent narcotic analgetic, and its unnatural (+)-antipode (4), were both poor inhibitors of the enzyme. (-)-Dihydrosecophysostigmine (5), a ring-open analog of (-)-physostigmine was less, but (-)-N-methylphysostigmine (6) much more potent than the natural alkaloid. The availability of compounds related to (-)- and (+)-physostigmine by improved chemical synthesis suggests that further structural variation may well lead to other biologically interesting AChE inhibitors.

Acetylcholinesterase↗

Smoking as a risk factor for recurrence of sudden cardiac arrest.

We obtained information about cessation of smoking in 310 survivors of out-of-hospital cardiac arrest who had been habitual cigarette smokers at the time of their arrest. Patients with coronary heart disease were stratified according to mortality risk on the basis of recognized criteria. The expected first-year rate of recurrent arrest ranged from 2 to 40 percent among the strata. Life-table analyses showed that reformed smokers had a lower incidence of recurrent arrest than patients who continued to smoke (19 vs. 27 percent at three years; P = 0.038 by one-sided test adjusted across strata). This effect occurred to varying degrees in all but the highest risk stratum. No differences in survival were observed for mortality due to other causes. It is possible that continued smoking in these patients led to acceleration of an ongoing atherosclerotic process, but the differences in early survival suggest that smoking may also act in the short term to enhance vulnerability to cardiac arrest.

Coronary Disease↗

Monoclonal antibodies reveal extensive antigenic differences between the hemagglutinin-neuraminidase glycoproteins of human and bovine parainfluenza 3 viruses.

A panel of twenty monoclonal antibodies to the hemagglutinin-neuraminidase (HN) glycoprotein of human parainfluenza 3 (PI3) virus has been obtained and tentatively classified into four different groups based on reactivity in hemagglutination inhibition (HI), neuraminidase inhibition (NI), and plaque neutralization (NT) tests. The antibodies were tested for cross-reactivity with bovine PI3 virus, Sendai virus, and simian virus 5 (SV5). Only two of these antibodies showed similar reactivities with human and bovine PI3 viruses in HI and NT tests; a few other antibodies showed low levels of reactivity with the heterologous viruses in HI tests. A competitive binding assay further suggested that the two cross-reactive antibodies are directed against the same domain of the HN molecule. Therefore, the HN glycoproteins of human and bovine PI3 viruses appear to be antigenically dissimilar, although they share at least one common epitope.

Antibodies, Monoclonal↗

Considerations for improving survival from out-of-hospital cardiac arrest.

Since the implementation of a paramedic system in Seattle, yearly survival rates from out-of-hospital cardiac arrest due to ventricular fibrillation have averaged 25% without any significant increase over the years. Outcome for cardiac arrest associated with other rhythms has been poor: when asystole was the first rhythm recorded, only 1% of patients survived; when electromechanical dissociation was initially present, only 6% survived. For cases of electromechanical dissociation, neither the type of rhythm nor the rate appear to influence outcome. Survival from ventricular fibrillation can be improved by shortening the delay to initiation of CPR and to defibrillation. When outcome in 244 witnessed arrests was related to the times to beginning CPR and to initial defibrillation, mortality increased 3% each minute until CPR was begun and 4% a minute until the first shock was delivered. New strategies that minimize delays appear to have the greatest promise for improving survival after cardiac arrest.

Ambulances↗

Factors influencing survival after out-of-hospital cardiac arrest.

Survival to hospital discharge was related to the clinical history and emergency care system factors in 285 patients with witnessed cardiac arrest due to ventricular fibrillation. Only the emergency care factors were associated with differences in outcome. Both the period from collapse until initiation of basic life support and the duration of basic life support before delivery of the first defibrillatory shock were shorter in patients who survived compared with those who died (3.6 +/- 2.5 versus 6.1 +/- 3.3 minutes and 4.3 +/- 3.3 versus 7.3 +/- 4.2 minutes; p less than 0.05). A linear regression model based on emergency response times for 942 patients discovered in ventricular fibrillation was used to estimate expected survival rates if the first-responding rescuers, in addition to paramedics, had been equipped and trained to defibrillate. Expected survival rates were higher with early defibrillation (38 +/- 3%; 95% confidence limits) than the observed rate (28 +/- 3%). Because outcome from cardiac arrest is primarily influenced by delays in providing cardiopulmonary resuscitation and defibrillation, factors affecting response time should be carefully examined by all emergency care systems.

Aged↗

Evaluation of a photolabile derivative of 1,25-dihydroxyvitamin D3 as a photoaffinity probe for 1,25-dihydroxyvitamin-D3 receptor in chick intestinal cytosol.

We evaluated the viability of 1 alpha, 25-dihydroxyvitamin D3-3 beta-[N-(4-azido-2-nitrophenyl)glycinate] (1,25-(OH)2-D3-ANG), an analog of 1 alpha, 25-dihydroxyvitamin D3 (1,25-(OH)2-D3) as a photoaffinity probe for 1,25-(OH)2-D3 receptor in chick intestinal cytosol. A competitive-binding assay revealed that chick intestinal cytosolic 1,25-(OH)2- D3 receptor bound to 1,25-(OH)2-D3-ANG approximately 20-times less effectively than it did to 1,25-(OH)2-D3. Irradiation of 1,25-(OH)2-D3- ANG in the presence of chick intestinal cytosolic preparation significantly diminished subsequent binding to 3H-1,25-(OH)2-D3, suggesting that the photoaffinity analog was covalently attached to the receptor. Therefore the nitroarylazide derivative of 1,25-(OH)2-D3 may be a valuable photoaffinity probe for the characterization of the 1,25-(OH)2-D3 receptor.

Affinity Labels↗

Somatic and excretory polysaccharide antigens of Leishmania donovani promastigotes.

Leishmania donovani promastigotes were found to release excretory-secretory antigens (ESA) in the culture medium which reacted against rabbit antiserum to live promastigotes (RA-LP). The active material was purified and found to contain mainly polysaccharides with about 23% of protein. A polysaccharide-rich antigen (PRA) was also isolated from the promastigote debris by the phenol-water extraction procedure. When tested against RA-LP in agar gel diffusion, both the preparations were found to be antigenically identical to each other as well as to a preparation of crude soluble antigen (CSA). The determinants involved were shown to be polysaccharide in nature. Gel filtration of all the three preparations showed that the antigenic activity was distributed in various eluted fractions. Both CSA and PRA also exhibited considerable electrophoretic heterogeneity in immunoelectrophoresis, although ESA showed only two overlapping faster moving bands. It is concluded that ESA represented a mixture of highly negatively charged mucopolysaccharides of varying sizes. The minimum molecular weight of the basic structural unit was around 2 X 10(4) daltons. These materials were probably derived from antigenically similar complexes of larger molecular weights located on the outer surface of the parasite and were released in the culture medium during their multiplication. Information available so far from this study suggests that the polysaccharide determinants of ESA/PRA did not elicit any marked immune response in kala-azar patients.

Animals↗

Glycoproteins of human parainfluenza virus type 3: characterization and evaluation as a subunit vaccine.

The envelope glycoproteins of human parainfluenza type 3 virus were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and reactivity with specific monoclonal antibodies. The molecular weight of the hemagglutinin-neuraminidase (HN) glycoprotein was found to be 72,000, and the fusion (F) glycoprotein appeared to consist of 74,000 (F0) or 56,000 (F1) species. Envelope glycoproteins were solubilized with octyl-glucoside and, after removal of the detergent by dialysis, were used for immunization of hamsters. Other animals were immunized with a formalin-inactivated preparation of whole virus. A single subcutaneous immunization with these antigen preparations induced a serum antibody response to the HN and F glycoproteins, as determined by plaque neutralization, hemagglutination inhibition, inhibition of virus-induced cell fusion, and immune precipitation tests. An IgG antibody response to both glycoproteins was also observed in bronchial washings. Animals immunized with the highest dose of envelope glycoproteins showed complete protection from challenge infection, whereas immunization with inactivated virus did not completely protect animals.

Animals↗

The influence of halothane and isoflurane on pulmonary collateral ventilation.

The effects of halothane and isoflurane on hypocapnic increases in pulmonary collateral resistance were studied in dogs. A bronchoscope with a double lumen catheter in the suction port obstructed a peripheral airway and allowed gas to flow out of the isolated segment of lung only via collateral channels. The collateral gas flow (Vcoll) was measured with a flowmeter and delivered through one lumen of the catheter, while the other lumen measured distal pressure (Pb). At FRC, the resistance to collateral ventilation (Rcoll) was calculated as Rcoll = Pb/Vcoll. The rest of the lung was ventilated with air, while air (hypocapnia), 10% CO2 in air, or air and halothane or isoflurane were delivered to the isolated segment. A measurement of resistance was made after 4 min of test gas flow. For each segment, when air replaced 10% CO2, the average increase in Rcoll was calculated and called Rmax. When 10% CO2 in air was infused into segments the mean Rcoll (n = 50) was 0.0196 +/- 0.0022 cmH2O X ml-1 X min. This increased to 0.0285 +/- 0.0031 cmH2O X ml-1 X min (mean +/- E) when air was infused, a mean increase in resistance of 52 +/- 3%. When halothane or isoflurane was added to air the hypocapnic increase in Rcoll was attenuated with a 50% decrease at 1.3% (1.4 MAC and 0.8 MAC, respectively). These two inhalational anesthetics reduce active changes in the flow resistance to collateral ventilation. When collateral resistance acts to adjust ventilation perfusion deviations, this action of halothane and isoflurane may make this regulation less effective.

Airway Resistance↗

Amplitude of ventricular fibrillation waveform and outcome after cardiac arrest.

The amplitude of ventricular fibrillation found initially in 394 patients was compared to clinical and logistical findings at the time of cardiac arrest. Peak-to-peak amplitude averaged 0.55 +/- 0.25 mV; a very low amplitude (0.2 mV or less) or "fine" fibrillation was present in 66 patients (17%). The amplitude was not found to be related to clinical histories, but depended on the length of the period from collapse until start of basic life support (p = 0.004) and the delay until assessment by paramedics (p = 0.001). Survival rates were strongly associated with amplitude: only 4 patients (6%) with fine ventricular fibrillation survived, compared to 117 or 328 patients (36%) in whom the initial amplitude was higher (p less than 0.001). Patient outcome related to amplitude even after adjusting for clinical history and logistical delays (p less than 0.005). We conclude that fine ventricular fibrillation is in part the result of delay in initiation of treatment, and that fibrillation amplitude is a powerful indicator of outcome after cardiac arrest.

Aged↗

Demographic factors associated with dialysis technique failures among patients undergoing continuous ambulatory peritoneal dialysis.

We evaluated factors that would predict a successful outcome on continuous ambulatory peritoneal dialysis. We found that poverty, the need for a helper to carry out the dialysis, and physician allocation to therapy was associated with a poorer technique success. Neither age, education, marital status, sex, rural home, nor the presence of diabetes were important risk factors by themselves.

Adolescent↗

Oral contraceptive use: prospective follow-up of women with suspected glucose intolerance.

593 women from a cohort of 8,652 healthy women failed a 1-hr glucose screening test. These 593 women with suspected glucose intolerance were classified as normal or as having impaired glucose tolerance (IGT) or diabetes mellitus (DM) after an oral standard glucose tolerance test (SGTT) and were followed over a mean 8.55 years. No permanent change in glucose status as a result of oral contraceptive use was found in an analysis for impaired glucose tolerance, diabetes mellitus, diabetic symptoms, or use of oral agents and/or insulin. Increased glucose intolerance was observed for women who were older, had a positive family history of diabetes, or were of greater relative weight. Diabetic symptoms correlated most closely with the onset of persistent fasting hyperglycemia.

Adolescent↗

A possible mechanism for diisopropylfluorophosphate-induced memory loss in rats.

Passive avoidance retention and cortical [H3]-quinuclidinyl benzilate (QNB) binding were examined in rats that were chronically treated with diisopropylfluorophosphate (DFP), an irreversible acetylcholinersterase inhibitor. Retention of a passive avoidance response in DFP-treated rats was significantly lower when compared to vehicle-treated controls. Passive-avoidance retention decreased from 93% in control animals to 68% in DFP-treated rats. QNB binding studies revealed the density of muscarinic receptors in cortical homogenates was significantly reduced from 0.95 +/- 0.04 pmole/mg protein in controls to 0.72 +/- 0.04 pmole/m protein in DFP-treated rats. Scatchard analysis of QNB binding curves did not reveal a decrease in affinity of muscarinic receptors for QNB. Based on data that DFP causes a reduction in cholinergic receptors, this study supports the hypothesis that central cholinergic receptors are associated with mechanisms involved in memory storage.

Animals↗

Improved neurologic recovery and survival after early defibrillation.

Eighty-seven patients who had out-of-hospital cardiac arrests received defibrillating shocks delivered by minimally trained first responders before the arrival of paramedics in a city with short emergency response times. Their outcomes were compared with those of 370 other victims who received only basic life support by first responders until paramedics arrived. Survival was improved by early defibrillation in cases in which there was a delay in initiating cardiopulmonary resuscitation and in which paramedic response times exceeded 9 min; there was 62% survival after early defibrillation by first responders and 27% if first responders provided only basic life support (p less than .02). Neurologic recovery was also improved after early defibrillation. Eighteen of 46 resuscitated patients (39%) receiving early defibrillation were awake at 24 hr compared with 49 of 204 patients (24%) who received only basic life support while awaiting paramedics (p less than .02). Incorporating defibrillation as part of basic life support can reduce both mortality and morbidity from cardiac arrest, even in cities with established, rapidly responding emergency care systems.

Allied Health Personnel↗