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Biomedical subjects

G Church

Publications and source records attributed to G Church.

At least 19 recordsLinked to original sources

Electrophore mass tag dideoxy DNA sequencing.

Toward a goal of dideoxy sequencing DNA utilizing electrophore labels, we prepared four electrophore-labeled DNA oligonucleotide primers. Each primer has a different electrophore and DNA sequence but a common glycol keto (alpha,beta-dihydroxyketo) release group. Cleavage of this latter group by either periodate oxidation or a thermal retroaldol reaction releases the electrophores for detection by mass spectrometry. Successful sequencing data with these primers was obtained by capillary electrophoresis on an ABI Model 310 after fluorescence dideoxy terminator cycle sequencing reactions were conducted. In a separate experiment, it was demonstrated that a cocktail of the four electrophore DNA primers could be detected as a dried sample spot by CO2 laser desorption/capillary collection/gas chromatography electron capture mass spectrometry. These results establish some feasibility for our long-term goal of high-speed multiplex electrophore mass tag dideoxy DNA sequencing. Ultimately we plan to use a higher number of electrophore mass tags and to rely on direct detection of the desorbed electrophores by electron capture time-of-flight mass spectrometry.

Chromatography, High Pressure Liquid↗

Detection via laser desorption and mass spectrometry of multiplex electrophore-labeled albumin.

Albumin was reacted with a mixture of six electrophore N-hydroxysuccinimide esters, each of which possessed an interior glycolketo linkage. The purpose of this linkage is to release the attached electrophore as a ketone when heated, due to a thermal retro-aldol reaction. The multiplex electrophore-labeled albumin was detected as a dried spot deposited on a polyimide membrane by laser desorption/capillary collection (of the released ketone electrophores)/off-line gas chromatography/electron capture-mass spectrometry. This encourages further study of such electrophore labels in immunoassays and related techniques, where there is a need to make advances in multi-analyte detection.

Albumins↗

Task-based estimation of past exposures to 60-hertz magnetic and electric fields at an electrical utility.

OBJECTIVES: Past exposures of electric utility workers to extremely low-frequency (ELF) magnetic (B) and electric fields (E) in Quebec were estimated. METHODS: The current intensities were measured and durations of exposures determined for tasks or work locations in 14 job categories. Past task or location intensities were extrapolated from the present on the basis of interviews with long-term workers and utility personnel. Past task or location durations were estimated for the long-term workers. Time-weighted average (TWA) exposures for past periods were reconstructed for jobs from the intensity and duration estimates. RESULTS: Magnetic fields were estimated to have increased the most over time for substation and distribution-line jobs. Magnetic field exposures for jobs in the generation and transmission of electricity were estimated to have increased very little. For substation jobs, the ratios of magnetic fields in 1945 to those in 1990 ranged from 0.42 to 0.69; the corresponding figures for distribution-line jobs ranged from 0.36 to 0.94. For electric fields in substations, the estimated increase over time was less than for magnetic fields, the 1945:1990 ratios ranging from 0.59 to 0.88. For the distribution line jobs, the 1945:1990 ratios for electric fields were less than 1.0 in 4 cases (0.6 to 0.89), more than 1.0 in 3 others (1.13 to 2.01) and unchanged in 1. CONCLUSIONS: Reconstruction of TWA exposures allowed changes in the intensity and the duration of exposures to be considered separately. Documentation of the intensity and duration of exposures for different tasks allows exposure reconstruction for jobs that have ceased to exist. The method is applicable elsewhere if exposure-monitoring records allow the level and duration of exposures for tasks or locations to be calculated and if estimates of past durations and intensities of exposures can be reliably obtained.

Electromagnetic Fields↗

Cloning and sequencing of thiol-specific antioxidant from mammalian brain: alkyl hydroperoxide reductase and thiol-specific antioxidant define a large family of antioxidant enzymes.

A cDNA corresponding to a thiol-specific antioxidant enzyme (TSA) was isolated from a rat brain cDNA library with the use of antibodies to bovine TSA. The cDNA clone encoded an open reading frame capable of encoding a 198-residue polypeptide. The rat and yeast TSA proteins show significant sequence homology to the 21-kDa component (AhpC) of Salmonella typhimurium alkyl hydroperoxide reductase, and we have found that AhpC exhibits TSA activity. AhpC and TSA define a family of > 25 different proteins present in organisms from all kingdoms. The similarity among the family members extends over the entire sequence and ranges between 23% and 98% identity. A majority of the members of the AhpC/TSA family contain two conserved cysteines. At least eight of the genes encoding AhpC/TSA-like polypeptides are found in proximity to genes encoding other oxidoreductase activities, and the expression of several of the homologs has been correlated with pathogenicity. We suggest that the AhpC/TSA family represents a widely distributed class of antioxidant enzymes. We also report that a second family of proteins, defined by the 57-kDa component (AhpF) of alkyl hydroperoxide reductase and by thioredoxin reductase, has expanded to include six additional members.

Amino Acid Sequence↗

Effect of competition on performance of thoroughbred racehorses.

The effect of competition and the influence of age and sex on performance were examined in a study of 18 Thoroughbred racehorses. The horses performed two solo and two competitive runs at 1,200 and 1,600 m for a total of eight runs. No group ran faster during competition, which may have been a reflection of the quality of horses used for this study and their susceptibility to stress-induced impairment of performance. Males showed no significant difference between competitive and solo run times, whereas females were consistently slower during competition. Males ran significantly faster than females in all runs. There was no difference in run times due to age, which may have been due to the high mean age (5.9 yr) of the group. The slower competitive run times may have occurred because of an earlier onset of fatigue when compared with solo runs. Plasma lactate was significantly greater for the 1,200-m competitive than for the solo runs.

Age Factors↗

Exercise induced hormonal and metabolic changes in Thoroughbred horses: effects of conditioning and acepromazine.

Nine Thoroughbred horses were assessed to determine the normal response of insulin, glucose, cortisol, plasma potassium (K) and erythrocyte K through conditioning and to exercise over 400 and 1,000 m. In addition, adrenaline, noradrenaline, cortisol, plasma K, erythrocyte K and L-lactate concentrations were evaluated in response to maximal exercise with and without the administration of acepromazine. Conditioning caused no obvious trends in plasma K, erythrocyte K, insulin or glucose concentration. Serum cortisol increased (P less than 0.05) from the initial sample at Week 1 to Weeks 4 and 5 (attributed to a response to training), and then decreased. During conditioning, three horses had low erythrocyte K concentrations (less than 89.3 mmol/litre). Further work is needed to define the significance of low erythrocyte K concentrations in the performance horse. In all tests maximal exercise increased plasma K, glucose and cortisol concentrations, whereas insulin and erythrocyte K concentrations decreased. Thirty minutes following exercise, plasma K and erythrocyte K concentrations returned to resting values; whereas glucose and cortisol concentrations continued to increase and the insulin concentration also was increased. The magnitude of the changes varied for pre-conditioned vs post-conditioned exercise tests and the duration of exercise. The administration of acepromazine prior to exercise over 1,000 m failed to alter the circulating noradrenaline and adrenaline concentrations in anticipation of exercise or 2 mins following exercise. Acepromazine administration, however, did cause lower L-lactate concentration 2 mins (P less than 0.03) and 30 mins (P less than or equal to 0.005) following exercise. Also, erythrocyte K showed a delayed return to baseline levels at 30 mins post exercise. Further evaluation of these trends may help explain the beneficial role acepromazine plays in limiting signs of exertional rhabdomyolysis when administered prior to exercise.

Acepromazine↗

Furosemide and sodium bicarbonate-induced alkalosis in the horse and response to oral KCl or NaCl therapy.

Metabolic alkalosis was induced in 10 clinically normal horses by administration of furosemide (1 mg/kg of body weight, IM) followed 4.5 hours later by sodium bicarbonate (NaHCO3; 500 g in 8 L water) via nasogastric tube. Furosemide diuresis resulted in a mean weight loss of 21.1 kg, which was associated with small, but significant, increases in venous blood pH, bicarbonate, and plasma protein concentrations (P less than 0.001), while plasma potassium, chloride, and calcium concentrations declined significantly (P less than 0.001). Oral administration of the hypertonic NaHCO3 solution resulted in clinical evidence of hypovolemia, which was accompanied by a marked increase (P less than 0.001) in plasma protein concentration. Seven of the 10 horses developed signs of neuromuscular excitability, as evidenced by muscle fasciculations, and 5 of the horses developed diaphragmatic flutter. Hypernatremia was transiently induced, but it resolved as the horses were allowed access to water. The alkalosis induced by furosemide and NaHCO3 was profound and persisted for a 24-hour period and was associated with marked hypochloremia and hypokalemia. Partial replacement of the electrolyte deficits and correction of the metabolic alkalosis was attempted, using 1,000 mEq of NaCl or KCl given as an isotonic solution via nasogastric tube. In the KCl-treated group, there was a prompt and significant decline in venous blood pH and bicarbonate concentration (P less than 0.001) accompanied by a significant increase in plasma potassium concentration (P less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Alkalosis↗

Influence of furosemide treatment on fluid and electrolyte balance in horses.

Alterations in electrolyte and acid-base balance were studied in 6 horses for 8 hours after furosemide administration (1 mg/kg of body weight, IM), and the results were compared with those for 5 healthy untreated horses (controls) kept under identical environmental conditions. In the treated group, decreases in plasma potassium, chloride, and calcium concentrations and increases in total plasma protein content persisted for the 8-hour observation period, whereas there was no change in plasma sodium concentration, osmolality, or packed cell volume. Plasma bicarbonate concentration and PCO2 remained high throughout the study, during which time venous blood pH was modestly increased only at the 6-hour sampling time. Furosemide treatment resulted in decreases in urine pH, specific gravity, osmolality, and potassium and calcium concentrations and increases in urine volume and total urine sodium, chloride, and calcium excretion. Body weight decreased 19.2 +/- 5.2 kg (mean +/- SD) in treated horses (4 +/- 1% of body weight), compared with a weight loss of 8 +/- 2.1 kg in untreated horses (1.5 +/- 0.4% of body weight) during the 8-hour experimental period. The increased fluid losses induced by the diuretic did not cause any obvious clinical signs in the horses. Pulse pressure, skin turgor, capillary refill time, and jugular distensibility remained unchanged throughout the experimental period.

Acid-Base Equilibrium↗

Effects of hypoxia and hyperoxia on ventilatory kinetics during recovery from exercise.

The purpose of this investigation was to determine the effects of breathing hyperoxic and hypoxic gas mixtures on ventilatory kinetics in the transition from submaximal exercise to rest. Eight male subjects performed three separate single blind exercise tests at 80% of their ventilatory threshold. Inspired oxygen concentration was varied in each experimental condition: test one (55% O2 -45% N2), test two (14% O2 -76% N2), and test three (21% O2 -79% N2). Ventilation, heart rate, and gas exchange were measured every 15 s for 6 min of exercise and during 9 min of recovery from exercise. Data analysis revealed no significant (p less than 0.05) differences in the kinetics of heart rate, oxygen uptake, expired volume of carbon dioxide, or ventilation among treatments during the transition from exercise to rest. Given the belief that hyperoxia attentuates the carotid bodies and hypoxia augments carotid body chemosensitivity, these findings suggest that the carotid bodies are not important regulators of VE kinetics during recovery from exercise.

Adult↗

Haemoglobin saturation during incremental arm and leg exercise.

There are few reports concerning the alterations in the percent of haemoglobin saturated with oxygen (%SO2) during non-steady state incremental exercise. Further, no data exist to describe the %SO2 changes during arm exercise. Therefore, the purpose of this study was made to assess the dynamic changes in %SO2 during incremental arm and leg work. Nine trained subjects (7 males and 2 females) performed incremental arm and leg exercise to exhaustion on an arm crank ergometer and a cycle ergometer, respectively. Ventilation and gas exchange measurements were obtained minute by minute via open circuit spirometry and changes in %SO2 were recorded via an ear oximeter. No significant difference (p greater than 0.05) existed between arm and leg work in end-tidal oxygen (PETO2), end-tidal carbon dioxide (PETCO2), or %SO2 when compared as a function of percent VO2 max. These results provide evidence that arterial O2 desaturation occurs in a similar fashion in both incremental arm and leg work with the greatest changes in %SO2 occurring at work rates greater than 70% VO2 max.

Arm↗

Swelling and ion uptake in cat cerebrocortical slices: control by neurotransmitters and ion transport mechanisms.

Cat cerebrocortical slices incubating in medium containing normal K+ concentrations were exposed to a number of different transmitters. Norepinephrine, histamine and adenosine or 2-chloroadenosine caused increased swelling of the slices associated with an increased Na+ and Cl- content. These effects were seen only when both Cl- and HCO3- were present in the medium, and were inhibited by a number of anion transport inhibitors. These characteristics were identical to those of the HCO3(-)-dependent component of the swelling induced by high K+ levels in the medium. Other transmitters, namely 5-hydroxytryptamine, dopamine, and gamma-amino butyric acid, were ineffective. The effects of norepinephrine, histamine and 2-chloroadenosine were antagonised by propranolol and phentolamine, chlorpheniramine and diphenhydramine, and theophylline respectively. These antagonists also inhibited HCO3(-)-dependent, K+-stimulated swelling. The transmitters which induced swelling also stimulated the carbonic anhydrase activity of cerebrocortical slices. We conclude from these data that the HCO3(-)-dependent component of K+-stimulated swelling may be due to K+-stimulated release of transmitters. Furthermore, the fact that the transmitters which induce swelling have also been reported to be most effective in increasing cAMP content in both brain slices or cultured astrocytes is consistent with the swelling response being mediated via cAMP-induced changes and being predominantly localized to astrocytes.

2-Chloroadenosine↗

Staffing the CCU.

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Arrhythmias, Cardiac↗

Triglyceride, insulin, and cortisol responses of ponies to fasting and dexamethasone administration.

Ponies were evaluated for their response to feed withholding and exogenous administration of corticosteroids (dexamethasone 0.04 mg/kg intramuscular [IM]) in an attempt to reproduce the hyperlipemia syndrome. Because insulin resistance has been associated with hyperlipemia, all ponies were initially evaluated for insulin response to an oral glucose load and normal dexamethasone suppression of serum cortisol. Four ponies were identified as hyperinsulinemic reflecting insulin resistance. All ponies had suppressed cortisol concentrations following dexamethasone administration. Feed withdrawal resulted in hypertriglyceridemia by 48 hours in all ponies. Very low density lipoprotein-triglyceride (VLDL) fraction was primarily elevated. The administration of dexamethasone failed to increase the degree of triglyceridemia. Although insulin resistance has been proposed as the likely cause of the hypertriglyceridemia in ponies, in this study four of eight ponies were considered to have normal insulin responses and yet still developed hypertriglyceridemia.

Animals↗