Oxidative enzymes in the urinary apparatus of several freshwater fishes.

The distribution and activities of several oxidative enzymes in the urinary apparatus of five freshwater fish species (river lamprey, lobe finned eel, Prussian carp, rainbow trout and three-spined stickleback) have been studied. Species were selected from three main taxonomic groups: Cyclostomata, Polypterini, Teleostei. Distinctly positive enzyme reactions were only found in the tubular elements of the kidney and the collecting duct-archinephric duct system, with the exception of the generally weak staining intensities of lactate dehydrogenase. The distal tubule normally showed strong to very strong reactions for most of the enzymes investigated. In the epithelial cells of the collecting tubule-collecting duct system, stronger reactions were observed for most of the mitochondrial-bound enzymes, especially succinate dehydrogenase and NADH-diaphorase. For these enzymes, the cells of the archinephric duct reacted strongly positive in Lampetra, Carassius and Gasterosteus. The enzyme patterns of various types of urinary tubules and ducts are compared with results of several morphological studies. In addition, the histochemical findings are discussed in relation to kidney function in different vertebrate groups.


[Therapy of traumatic tympanic membrane perforations (author's transl)].

Immediate operative reposition of traumatic eardrum perforations has been found to give better morphological results when compared with spontaneous healing. The operative technique and results of healing in 32 patients treated at the ENT Department, University Hospital, Zürich, are discussed.


A model of the effects of respiration on left ventricular performance.

We have investigated the interaction of left ventricular afterload and right-heart volume on left ventricular performance. By utilizing a right-heart bypassed heart-lung preparation, we have been able to control independently each factor. We have previously suggested that the major influences of the inspiratory decrease in pleural pressure in decreasing left ventricular stroke volume (LVSV) may be via the inspiratory increases in left-heart afterload and right-heart volume (RHV). Thus, our preparation serves as a model to study in detail the specific effects on the left ventricle of changes that are induced by respiration. Our results show that increases in transmural (relative to pleural pressure) aortic pressure (Pao) caused an increased left ventricular end-diastolic pressure (LVEDP), and that the effect of equivalent increases in Pao was greater when RHV was greater. Increase in RHV had minimal effects at low volume, but resulted in large increases in LVEDP at large RHV. These effects were markedly attenuated after pericardiectomy. Rapid increases in either RHV or Pao produced transient falls in LVSV. Our results are consistent with the hypothesis that increases in both right-heart volume and LV afterload contribute to the inspiratory decrease in LVSV and increase in LVEDP.


A continuum mechanics analysis of pulmonary vascular interdependence in isolated dog lobes.

The pressure-diameter (PD) behavior of intact pulmonary vessels was measured roentgenographically at several fixed deflation transpulmonary pressures (Ptp). At any constant vascular pressure (Pv), the intact-vessel diameter was larger for higher Ptp. The behavior of the parenchyma was described by a nonlinear analysis. An excised-vessel PD behavior was computed which was consistent with both nonlinear parenchymal behavior and measured intact-vessel PD behavior. Estimates of Px', the difference between the perivascular pressure and the pleural pressure, as a function of Ptp and Pv were obtained. For physiological values of Pv, mean values of Px' for arteries were -1 cmH2O at Ptp of 4 cmH20 and decreased almost linearly to -15 cmH20 at Ptp of 25 cmH2O. Veins showed a similar behavior, but Px' was more positive. Increasing Pv raised Px' and decreasing Pv lowered Px' at all values of Ptp. These results indicate that the interdependence effect is small at functional residual capacity, increases with lung inflation, and is greater for arteries than veins.


Circulatory regulation during exercise in different ambient temperatures.

Three relatively fit subjects performed duplicate 20- to 25-min cycle ergometer exercise bouts at moderate and heavy intensities (40% and 70% Vo2 max) in ambient temperatures of 20, 26, and 36 degrees C. They approached a steady state in internal body temperature (Tes) in all but the heavy exercise in the heat, where Tes rose consistently, averaging 38.84 degrees C at the termination of exercise. Cardiac output (Q), estimated by a rebreathing technique, was proportional to Vo2 and independent of the body temperatures, except during the lower exercise intensity in the heart, where Q averaged 1.31 . min -1 higher throughout. In any environment, forearm blood flow was linearly related to Tes above the Tes threshold for vasodilation, but during heavy exercise in the heat this relationship was severely attenuated above a Tes around 38.0 degrees C, when forearm blood flow exceeded 15 ml.min -1 .100 ml -1. Plasma volume decreases during exercise were primarily a function of the intensity of exercise. During heavy exercise in the heat the relative vasconstriction contributes to the maintenance of an adequate stroke volume preventing a fall in Q. In this case, circulatory regulation has precedence over temperature regulation.


Reversal of 2-deoxyglucose inhibition of serotonin uptake in isolated guinea pig lung.

The effect of various substrates on the rate of uptake of 5-hydroxytryptamine (serotonin) from the pulmonary circulation was studied with isolated guinea pig lungs. Lungs were ventilated and were perfused with an electrolyte solution in a recirculating system. Uptake of serotonin was calculated from the rate of disappearance of [14C]serotonin from the perfusate. Serotonin uptake was inhibited by approximately 40% when 5 mM 2-deoxyglucose was substituted for glucose in the pulmonary perfusate. The inhibition due to 2-deoxyglucose was reversed by addition to the perfusate of pyruvate, lactate, acetate, alanine, beta-hydroxybutyrate, palmitate, or glycerol. Fructose ans succinate were less efficient substrates while no significant effect was noted with L-glycerol 1-phosphate or L-phenylalanine. These results indicate that a wide variety of metabolizable substrates can be utilized by the pulmonary endothelium to support uptake of serotonin.


Muscle strength and speed of movement in relation to age and muscle morphology.

Maximum values for isometric strength, dynamic strength, and speed of movement (MEV) in the quadriceps muscle were measured in 114 male subjects who were between 11 and 70 yr. Biopsy samples were taken from the quadriceps muscle in 51 of the subjects (22-65 yr. old). Isometric and dynamic strength increased up to the third decade, remained almost constant to the fifth decade, and then decreased with increasing age. However, no measurable external atrophy of the quadriceps muscle, explaining the decline in strength, could be seen in old age. Histochemical changes in the muscle tissue such as decreased proportion of type II fibers and a selective atrophy of type II fibers, were seen with increasing age. The strength decline in old age was also observed to correlate significantly with the type II fiber area. Multiple regression analyses indicated, however, that mechanisms other than the type II fiber atrophy might be responsible for the decline in strength performance during aging. The implications of these findings are discussed.


Responses of forearm blood flow to graded leg exercise in man.

To test whether the cutaneous vascular responses to exercise are influenced by the level of work, three strategies were followed. In each, forearm blood flow and esophageal temperature were measured throughout. In part I, the forearm blood flow-internal temperature relationships from separate sessions of steady-state exercise at different loads were compared. In part II, work load was varied between 50 and 150 W. Work load was raised or lowered 50 W every 5 min over 60-75 min. The third strategy was to examine the immediate change in forearm blood flow accompanying rapid, large increments or decrements in work load. The results do not support a graded response of the cutaneous circulation to exercise. In both the first and second protocols above, the relationship of forearm blood flow to internal temperature was not measurably altered by work load. Multiple linear regression analysis failed to reveal a consistent role for work load in part II. In the third protocol, there was no consistent or sustained response to an abrupt change in work load. Thus over the range of work loads used in this study there appears to be no major role for the level of work in the regulation of forearm skin blood flow other than through the effect on internal temperature.


Flow-volume curves of excised right and left rabbit lungs.

Static pressure-volume (PV) curves and flow-volume (VV) curves of excised right and left rabbit lungs obtained by forced and passive deflation were compared. Deflation PV curves in which the volume was expressed as either ml/lung weight or percent total lung capacity were nearly identical between right and left lungs. Descending limbs of the forced VV curves in which the flow divided by vital capacity (VC) was plotted against %VC generally agreed between right and left lungs, although peak flow tended to be higher in left lungs. However, the flow obtained during passive deflation was higher in left lungs over most of the deflation suggesting that the resistance of proximal airways per unit volume is lower in left than in right lungs.


Role of respiratory heat exchange in production of exercise-induced asthma.

We have hypothesized that it is the total heat flux in the tracheobronchial tree during exercise that determines the degree of postexertional obstruction in asthma, and have developed quanititative expressions that relate these two events. We tested this hypothesis by comparing the observed responses to exercise, while our subjects inhaled dry air at various temperatures ranging from subzero to 80 degrees C in a random fashion, to those that we predicted would occur based upon calculations of respiratory heat exchange. We further determined if heat could be transferred from the inspired air to the mucosa so as to offset evaporative losses from the airways. The observed responses fell as air temperature was increased from -11 to +37 degrees C and exactly matched theoretical predictions. Above 37 degrees C, the observed response exceeded predictions, indicating that it was not possible to provide sufficient heat per se in the air to offset the vaporization of water. However, when small amounts of water vapor were added to the inspirate at high temperatures, bronchospasm was virtually abolished and the response again closely matched theoretical expectations. We conclude that the magnitude of exercise-induced asthma is directly proportional to the thermal load placed on the airways and that this reaction is quantifiable in terms of respiratory heat exchange.


Esophageal temperature during exercise in asthmatic and nonasthmatic subjects.

We measured the temperature in the rectum and two esophageal sites (retrocardiac and retrotracheal) during exercise in eight asthmatic and six normal subjects while they breathed air at subfreezing, ambient, and body conditions. Various aspects of pulmonary mechanics were recorded before and after exercise. The asthmatic subjects developed the greatest airway obstruction following exercise with cold air and no response at body conditions. There were no changes in pulmonary mechanics in the postexercise period in the normal individuals with any of the inspired air conditions. Despite these divergent mechanical responses retrotracheal temperatures fell by the same magnitude below core values in both groups of subjects, indicating that identical degrees of airway cooling occurred. We conclude that rather than having a defect in the ability to condition inspired air, asthmatic subjects are more responsive to the effects of incompletely conditioned air.


Body temperature and ventilatory responses to CO2 during chronic respiratory acidosis.

During acute hypercapnia (5% carbon dioxide) in resting conscious dogs, ventilation (Ve) attained a new level above control within 5 min, but rectal temperature decreased gradually to reach a steady state lower than control after 40-60 min. At 2 days of breathing 5% carbon dioxide, Ve remained elevated, as in acute hypercapnia, but Paco2 increased and the threshold of the ventilatory response shifted to a higher Paco2. By 2 days of hypercapnia, rectal temperature (Tr) had returned to normal, reflecting an alteration of hypothalamic temperature control that might be expected to result in enhanced respiratory drive. Surprisingly, despite blood acid-base compensation between 2 and 14 days of hypercapnia, Ve did not decrease, whereas Paco2 decreased to the level observed during acute hypercapnia, and the threshold of the ventilatory response returned to normal. Therefore, at 14 days of respiratory acidosis, acid-based compensation resulting from increase in bicarbonate was not associated with reduced respiratory drive. This result could not be accounted for on the basis of a temperature mechanism because temperature adaptation occurred earlier.


Pulmonary microvascular response to hemorrhagic shock, resuscitation, and recovery.

We studied the effect of hemorrhagic shock, resuscitation, and recovery on the pulmonary microcirculation. We used lung lymph flow (QL) and lymph-to-plasma protein ratio as sensitive indices of transvascular fluid filtration rate and protein permeability. We measured pulmonary vascular pressures, cardiac output, blood gases, lymph flow, and lymph and plasma proteins before and during a 2-h period of shock, a 3-h period of resuscitation, and a 72-h period of recovery, in nine unanesthetized sheep with chronic lung lymph fistula. We found a 30% decrease in QL during early shock as animals were bled into bags containing an acetate citrate dextrose solution until aortic pressure was 50 Torr. QL gradually increased to or exceeded base line in five of nine animals during late shock as pulmonary vascula resistance increased by 250%. During the 3-h resuscitation period, mean QL increased by 110%, with the lymph-to-plasma protein ratio being significantly decreased, indicating no protein permeability change. In five of nine studies, lymph became visible bloody. The increased QL and lymph RBCs were felt to be secondary to an elevation in microvascular pressure. During the recovery period, pressures and QL returned to base line.


Effect of pentobarbital anesthesia on rheology and transport of canine tracheal mucus.

Tracheal mucus samples were collected from six tracheostomized dogs before and after induction of pentobarbital anesthesia. The dynamic mechanical properties of the mucus were determined by the magnetic microrheometer technique. In 15 experiments, the mean value of elasticity at 1 rad/s rose from 67 dyn/cm2 before anesthesia to 212 dyn/cm2 in the period 15-90 min after induction. At the same time, mean viscosity at 1 rad/s rose from 24 to 79 P. The mucociliary transportability of some of the mucus samples was assayed by means of in vitro frog palate technique. In 10 experiments, the relative transport rate for dog tracheal mucus went from 98% of frog palate control befor anesthesia to 79% during pentobarbital anesthesia. A negative relationship between from palate transport rate and mucus elasticity was found. A negative correlation between transport and mucus viscosity at constant elasticity was also discovered. Finally, the mucus collection rate was reduced by a factor of about 8 during anesthesia, suggesting a reduction in the average depth of mucus.


A comparison of exercise responses in stairclimbing and cycling.

A free-paced 14.8-m stairclimb was compared to cycle ergometry at equivalent power outputs in six normal subjects. Heart rate (HR), inspiratory flow (VI) andd oxygen saturation in arterial blood (Sao2) were analyzed continuously, carbon dioxide output (Vco2) and oxygen consumption (Vo2) for the duration of exercise, and rebreathing mixed venous carbon dioxide pressure (Pvco2), capillary blood gases, and lactates at the completion of exercise. The average power output was 1,394 kpm/min sustained for 54.8 s. The HR and Vo2 responses were not significantly different between the two forms of exercise. VI was 20.3% less stairclimbing (P smaller than 0.005). A fall in Sao2 of 31.1% +/- 2.7) occurred in stairclimbing compared to 1.2% (+/- 2.1) in cycle ergometry. Differences in Sao2 between stairclimbing and cycling correlated with differences in VI (r = 0.80); falls in Sao2 could be prevented by overbreathing during stairclimbing or induced by controlled underbreathing during cycling. Vco2 and postexercise lactate levels were less stairclimbing (P smaller than 0.005 and P smaller than 0.01), as were Pvco2 measurements. The findings are consistent with a lower CO2 flow to the lung during stairclimbing than in cycling, which results in a lower alveolar ventilation and a consequent fall in Sao2.


Human skin wettedness and evaporative efficiency of sweating.

Rates of evaporation and sweating were recorded for three acclimatized male subjects in hot humid conditions, the ambient parameters of which were set so that the various imposed evaporative rates required the same skin wettedness at different levels of sweating. Rectal and skin temperatures were measured. Results showed that during steady state occurring during the 2nd h of exposure each subject reached the required evaporative rate by means of increases in skin wettedness regardless of the level of sweating; the sweat evaporative efficiency, defined as the ratio between evaporative rate and sweat rate, decreased as skin wettedness increased, in a range between 0.74 and 1.0 Sweat efficiency fell to 0.67 for fully wet skin. The body temperatures did not increase with time if skin wettedness was less than unity. Evaporative heat transfer coefficient (he), maximum evaporative capacity, and wettedness were estimated on the basis of the observed decrease of sweat efficiency. The relationship between skin wettedness and sweat efficiency was interpreted as a combined effect of differences in local he as well as in local sweat rates.


Pulmonary hypoxic vasoconstriction: not affected by chemical sympathectomy.

The influence of chemical sympathectomy with 6-hydroxydopamine (6-OHDA) on regional alveolar hypoxic vasconstriction and on global hypoxic pulmonary vasoconstriction was investigated. In eight dogs a double-lumened endotracheal tube allowed ventilation of one lung with nitrogen as an alveolar hypoxic challenge while ventilation of the other lung with 100% O2 maintained adequate systemic oxygenation. Distribution of perfusion to the two lungs was determined with 133Xe and external counters. Mean perfusion to the test lung was 50.9 +/- 4.9% of total lung perfusion on room air and decreased by 32.4% (P smaller than 0.01) during alveolar hypoxia. Following 6-OHDA the test lung continued to reduce perfusion during alveolar hypoxia by 27.3%. In five dogs global hypoxia induced a 106% increase in pulmonary vascular resistance (PVR) prior to 6-OHDA and a 90% increase in PVR after 6-OHDA. After 6-OHDA no rise in PRV or systemic blood pressure occurred in response to tyramine, confirming effective sympathectomy by the 6-OHDA. Thus, sympathectomy with 6-OHDA failed to substantially block regional alveolar hypoxic vasoconstriction or global hypoxic pulmonary vasconstriction.


Mean alveolar gases and alveolar-arterial gradients in pulmonary patients.

In view of uncertainties about the best way to estimate mean alveolar gases in patients with ventilation-perfusion inequalities, three different methods were evaluated on 54 patients. 1) O2 and CO2 were recorded by mass spectrometer on an O2 (x)-CO2 (y) diagram. The coordinates at the intersect of the expiratory record with the mixed expired R line (RE) ives the mean alveolar values (PAo2 and PAco2. 2)pa'co2 was calculated with the Bohr equation using a predicted anatomic dead space and PA'o2 was derived with the alveolar equation. 3) End-tidal (ET) P02 were averaged over 1 min at rest in steady state. Mean RET calculated from 3 was identical with RE. Mean values for PAco2, PA'CO2. and PETco2 differed by less that 1 Torr, but the variance was least with the end-tidal method. There was a highly significant correlation between delta aAPco2 using PETco2 and VD/VT, better than with either of the other methods. The end-tidal measurement appears to give the best approximation of mean alveolar gas in pulmonary patients.


Cardiac responses of dogs to nonsynchronous and heart synchronous whole-body vibration.

Changes in cardiac function produced by synchronizing vibration-induced forces with events in the cardiac cycle were compared to those for the nonsynchronous case in eight chronically instrumented, tranquilized dogs. The supine animals received sinusoidal, whole-body vibration (along the spinal +/- Gz axis) at a constant acceleration amplitude (+/- 0.75 G). The vibration frequency (2-3 Hz) was set equal to the paced heart frequency. Synchronization between vibration and cardiac cycles on a beat-by-beat basis produced a specific and sustained cardiovascular response; such a sustained response was not possible with nonsynchronous vibration. With synchronization, relationships could be found and sustained in which changes either exceeded or were below mean nonsynchronous levels, in some cases below previbration values. For the synchronous vs. nonsynchronous states, significant modification of cardiac function was reflected in parameters such as coronary flow (+15% to -34%) and myocardial oxygen consumption (+21% to -51%). Thus, whole-body oscillation acceleration (vibration) is a forcing function that can produce and maintain a particular cardiovascular response.


Sex and age differences in intrathoracic airways mechanics in normal man.

Expiratory pressure-volume curves and maximal expiratory flow-volume curves were obtained in 74 healthy subjects (49 males 25 females) aged between 20 and 64. Maximal expiratory flowstatic recoil (MEFSR) curves were then constructed. Aging was associated with loss of lung recoil and reduction in maximal expiratory flow measured between 80 and 50% total lung capacity (TLC), in both males and females, with the slope of the MEFSR curve becoming steeper. In subjects less than 40 yr old the conductance of the upstream segment increased from 80 to 50% TLC, whereas in older subjects it decreased. We also studied the effects of alveolar gas compression artifacts on MEFSR curves in 12 additional subjects and 10 patients with chronic obstructive lung disease; consistent changes were found, but the subsequent shift of the MEFSR curve toward the left was only mild. These changes are interpreted as reflecting the net effects of an increase in the unstressed dimensions of the airways together with a decrease in intraparenchymal airways diameter, probably due to loss of parenchymal airways diameter, probably due to loss of parenchymal support. It was also concluded that the analysis of the MEFSR curve did not allow a quantitative estimate of the chantes in airways compressibility with aging.


Variability of the configuration of maximum expiratory flow-volume curves.

With a computer technique variability of the configuration of maximum expiratory flow-volume (MEFV) curves was studied in terms of slope ratio, SR. SR = dV/dV divided by V/V, where V is the instantaneous flow and V is the volume increment above residual volume.) Approximately four SR-V curves, each based on three to five smoothed and averaged MEFV curves, were derived for each of 20 normal subjects (aged 23-55 yr) on a single occasion, and again at least 1 wk later. Individual curves were largely reproducible, the maximum difference in SR at most volumes being 0.3-1 U, but frequently showed substantial yet reproducible fluctuations with volume. These corresponeded to hitherto unrecognized irregularities of maximum expiratory flow that may reflect sudden changes in the location of flow limitation.


Information content of the multibreath nitrogen washout.

The distribution of ventilation per unit volume (Va/Vol) in the lung is often studied using the multibreath N2 washout test. Most approaches have focused on the determination of a single Va/Vol distribution (consisting of from two compartments to a continuum) compatible with the data, but because the potential number of lung units greatly exceeds the number of breaths measured, many distributions are usually compatible with a given set of washout data. Interpretation of single distributions therefore requires evaluation of the variability among all such compatible distributions. The technique of linear programing is well suited to evaluation of these compatible distributions and its application to the N2 washout is explained. Twelve examples of washout data are analyzed in this way. The results indicate that using the first 20 or so breaths of the washout, narrow distributions of Va/Vol are well specified. Distributions with up to four distinct modes are also well specified, but for broad distributions the shape cannot be adequately defined. Good resolution in the region of high Va/Vol is found to be critically dependent on measuring the first few breaths of the washout.


Stroke volume and systolic time interval adjustments during bicycle exercise.

By use of the impedance noninvasive method, cardiac output (Q), stroke volume (SV), heart rate (HR), Heather index (HI), and systolic time interval (STI) values were studied in 17 subjects working at 25, 50, and 75% of their maximal oxygen uptake (Vo2 max) on a Monark bicycle. A significant increase in SV at each work load and a concomitant decrease in positive expiratory pressure (PEP), left ventricular ejection time (LVET), and PEP/LVET were observed. The linear regression equation at 75% Vo2 max between LVET and HR and SV confirms the close relationship between these parameters. The significa-t increase of the contractility index (HI) showing the stress response of the myocardium is clearly assessed. At 75% work load, its correlation with SV is low; but the high level of the correlation (r = 0.90) between HI and HR in the stress response. Consequently the changes in STI and HI provide valuable information on SV adjustment during calibrated effort responses. Consequently, the changes in STI and HI, the relationship between STI and SV, and the study of SV, HR, and Q provide valuable information about the adjustment of these parameters during calibrated efforts.


Effects of pulmonary circulation on blood levels of insulin and glucagon in ambulant dogs.

The effect of the lung on insulin and glucagon were studied by comparing aortic and mixed venous levels in ambulant dogs with indwelling catheters. In the resting state ther was no difference for either hormone. After injection of insulin aortic levels exceeded mixed venous for 10 min; mixed venous levels exceeded aortic for the next 36 min. After injection of glucagon mixed venous levels consistently exceeded aortic. Single-pass studied showed an apparent gain in immunoreactive insulin as compared with [14C]inulin and 125I-albumin during passage through the lung; but no effect on glucagon could be demonstrated. A loss of immunoreactive hormone across the lung might be explained by degradation within the capillary lumen be endothelial cell surface peptidases or uptake onto specific receptors. The apparent gain of immunoreactive insulin across the lung might be due to displacement of immunoreactive exogenous hormone competing for receptor sites or modification of exogenous hormone by the lungs with increase in immunoreactivity.


A method for continuous measurement of ammonia in respiratory airways.

A method has been developed that can continuously measure respiratory NH3 with a detection limit of 30 parts per billion by volume (ppbv) at an averaging time of 1 s. The method is based on the chemiluminescent detection of NO, which is produced by oxidizing NH3 at 850 degrees C. A polyethylene sampling probe, incorporating a heated capillary tip bathed in dry, NH3-free diluent air, minimizes NH3 losses due to absorption in breath condensate. In one subject, flow and tidal volume were monitored during quiet mouth breathing while NH3 and CO2 were sampled at the lips. In 10 subjects, NH3, and CO2 were sampled at the lips during quiet mouth breathing ranged from 280 to 1,280 ppbv (mean plus or minus SE, 540 PLUS OR MINUS 85 PPBV).


Granuloma formation associated with subconjunctival injection of a corticosteroid in dogs.

In three dogs given a subconjunctival injection of methylprednisolone acetate for the treatment of ophthalmic disorders, a yellow mass developed at the injection site. Microscopic examination of each excised mass revealed circumscribed granulomatous inflammation surrounding a central core of amorphous foreign material.


Aortic interruption in a dog.

A 3-month-old English Bulldog was found to have a discontinuity between the ascending and descending aorta. There were no signs of illness associated with the interruption. The descending aorta was supplied with blood by collateral circulation. An aortic graft of woven synthetic material was surgically implanted to connect the ascending and descending aorta. Without any restrictions on the dog's activity, the dog has lived for over 1 year and continues to do well.


Characterization of renal defects in dogs with a syndrome similar to the Fanconi syndrome in man.

Ten adult dogs with multiple spontaneous defects of renal tubular reabsorption were studied. Clinical signs included polydipsia, polyuria, and glycosuria for 2 to 12 months. Eight of the dogs were Basenjis. Urinalyses revealed hyposthenuria, glycosuria, and amino aciduria in most dogs. Renal function was normal in 5 dogs and slightly reduced in the remainder. Moderate metabolic acidosis had developed in 3 dogs. Renal clearance studies revealed reduced tubular reabsorption of glucose, phosphate, sodium, potassium, and uric acid. Abnormal glucose tubular maximal curves were found. Results of oral glucose tolerance tests were normal. Two patterns of abnormal amino aciduria were evident: generalized amino aciduria and a pattern similar to that of cystinuria in dogs. Radiography of long bones and bone densitometry did not reveal any skeletal abnormalities. Five of the dogs died within 90 days of diagnosis; death was due to acute renal failure associated with profound dehydration, acidosis, and papillary necrosis. The other dogs remained stable without treatment after 18 months. Histopathology of kidneys did not reveal uniform abnormalities; some dogs had variable and nonspecific changes and others were normal. Electron microscopy did not reveal ultrastructural abnormalities in renal tubular cells. It was concluded that the syndrome in these dogs represents a new entity of renal disease in dogs, similar to idiopathic Fanconi syndrome in man.


Dissolution of a struvite nephrolith in a dog.

A large radiodense calculus in the left renal pelvis of a 22-month-old, male Great Dane disappeared one month following surgical removal of two struvite (magnesium ammonium phosphate) calculi from the right renal pelvis. The dog's urine likely became undersaturated with struvite for a sufficient period to permit dissolution of the renal calculus. Several factors may have contributed to the decrease in urine struvite concentration, including eradication of a urease-producing Proteus sp from the urinary tract and induction of polydipsia and compensatory polyuria by oral administration of sodium chloride.


Nondiabetic emphysematous cystitis in two dogs.

Emphysematous cystitis unassociated with diabetes mellitus was diagnosed in two dogs. One dog had chronic recurring urinary tract infection with aerobic and anaerboic bacteria, and the other had urinary tract infection associated with renal glucosuria. In both dogs, the cystitis was resolved with appropriate antibacterial therapy.


Sudden death in young dogs with myocarditis caused by parvovirus.

Sudden death of pups in the 4- to 6-week age range has recently been occurring in western Canada as a result of severe, primary, nonsuppurative myocarditis. At necropsy, the prominent macroscopic lesion was pulmonary edema, and microscopically, characteristic intranuclear inclusion bodies were found within cardiac myofibers in association with myocarditis. Ultrastructurally, numerous small particles resembling parvoviruses were found within the intranuclear inclusion bodies, which were positive by direct fluorescent antibody test for canine parvovirus. Of three pups inoculated with homogenate from affected myocardium, one developed lesions resembling canine parvoviral enteritis.


Myocarditis of probable viral origin in pups of weaning age.

Myocarditis in 4- to 8-week-old pups from 10 litters was characterized by sudden death. Histopathologic findings included mononuclear cellular infiltration and interstitial fibrosis in the myocardium of the left ventricle. Basophilic intranuclear inclusion bodies were seen in myocardial cells in 4 of 18 pups necropsied, suggesting a viral origin of the disease. Other pathologic changes were variable, but all were attributable to cardiac failure. Of 8 surviving pups examined, 7 had evidence of cardiac failure, including pulmonary edema, cardiomegaly, and cardiac arrhythmias.


Canine hyperadrenocorticism: pretreatment clinical and laboratory evaluation of 117 cases.

The physical findings, clinical signs, age, breed and sex distributions, and laboratory data of 117 dogs with untreated Cushing's syndrome were reviewed. Poodles, Dachshunds, and Boxers of all ages were found to be at increased risk, as were dogs of all breeds greater than or equal to 6 years old. Polydipsia, polyuria, progressive bilaterally symmetric alopecia, and abdominal distention were the most frequently observed clinical signs and physical findings. Lymphopenia, eosinopenia, above normal values of serum alkaline phosphatase, serum cholesterol, and sulfobromphthalein dye retention, and below normal urine specific gravity were the most frequent abnormalities found in the laboratory data. About 50% of the dogs had urinary tract infections. Final diagnosis was established on the basis of abnormally high plasma corticosteroid values in response to an intramuscular injection of adrenocorticotropic hormone.


Phycomycosis in fifteen dogs and two cats.

Phycomycosis was confirmed by histologic examination of biopsy specimens from 15 dogs and at necropsy in 2 cats. The fungal infections occurred most frequently in young adult dogs (1-3 yr) of the larger breeds. The gastrointestinal tract was the organ most commonly involved. Treatment by surgical excision, amphotericin B, and sodium iodide alone or in combination was attempted in five cases. Mortality was 100%.


Meningiomas in cats: a retrospective clinical study of 36 cases.

Thirty-six cases of meningiomas in cats were reviewed as to the clinical signs and anatomic location. Age, breed, and sex also were noted. It was found that, in general, meningiomas affected older cats, with two thirds of them being greater than or equal to 10 years old. There was no breed predilection, although there was a majority of male cats. In most cases, the anatomic location was supratentorial, with only one case being infratentorial, and no tumors were below the foramen magnum. A number of cats had multiple maningiomas, and this often complicated the anatomic localization by neurologic examination. Tentorial herniation (6 cases) was associated with positional nystagmus or tetraparesis, or both, in addition to the deficits caused by the tumor.


Cystic lymphangioma in a colt.

A 6-month-old Morgan colt was examined because of preputial edema and multiple nodular masses in the left inguinal region. A tentative diagnosis of lymphatic hamartoma was made on the basis of initial biopsy findings. The prognosis for surgical excision was poor due to the progressive expansile growth of the tumor. Secondary infectious complications developed, and the colt was euthanatized at the owner's request. An abdominal and retroperitoneal mass was seen at postmortem examination. The mass was adherent to the left lumbar musculature, the craniodorsal aspect of the pelvic inlet, the midjejunum, the left testicle, and the left kidney. The mass extended through the left inguinal and femoral canals into the deep intermuscular fascia of the hindlimb. Histologic examination of the mass indicated that it was a cystic lymphangioma.


Surgical decompression for traumatic atlantoaxial subluxation in a weanling filly.

A filly with ataxia and splinting and crepitation in the neck was found to have atlantoaxial subluxation. Radiographic diagnosis was based on the same criteria as those used in other species, ie, increased distance between the atlas and spine of the axis and increased distance between the dens and floor of the atlas. Extensive hemilaminectomy was performed to decompress the spinal cord. Stabilization was not attempted. Immediate postoperative response was encouraging, but the untimely death of the filly prevented further evaluation of the procedure.


Ultrasonic diagnosis of ocular pathology: its significance for cataract surgery with and without IOL implantation.

The use of diagnostic ultrasound can alert the eye surgeon to certain pathologic conditions of the eye before cataract surgery, including problems associated with the vitreous, retina, choroid and lens. This knowledge may be applied to all cataract surgery and in particular to intraocular lens implantation.


Production and life span of cutaneous mast cells in young rats.

Tritiated thymidine was injected into young rats (age 7 days) in such a way as to be incorporated into the nuclei of all cells that were synthesizing DNA during a period of 22 hours. Specimens of the skin of the back and the pinna of the ear were taken at intervals from one to 112 days after injection of the [3H]thymidine. The mast cells were stained with Alcian blue, and autoradiographs were prepared. The nuclei were counterstained with alum-brazilin. Making due allowance for growth of the animals, and for shrinkage due to histological preparation, the total numbers of mast cells in the dorsal skin and in the pinna were determined. The numbers of mast cells containing [3H]thymidine were calculated from the proportions of those cells found to have radioactive nuclie. Using these data, the rates of appearance of labelled mast cells, and of decline in their numbers with time were determined for both regions of skin. No mitotic figures were seen in any mast cells. It is concluded that mast cells arise by the division of agranular precursor cells of unknown identity. The characteristic cytoplasmic granules appear to be produced by the daughter cells during the 24-48 hours following premitotic replication of DNA in the precursors. The differentiated cells have half-lives of 4-9 days in the skin of the back and 7-20 days in the external ear. All the labelled mast cells had disappeared after 28 days in the back and after 84 days in the ear. Comparison of these findings with the results of other investigators suggests that the mast cells produced early in life have much shorter life spans than do most of the mast cells present in adult rats, The longer life span found in the pinna may account for the greater density of the cells there than in the back. This regional difference may reflect the greater need for mast cells in a region which is more susceptible to adverse environmental influences.


The small luteal cell of the sheep.

Corpora lutea of sheep were examined by electron microscopy at day 10 of the oestrous cycle and at days 15, 25, 50, 100, 125 and 140 of pregnancy. Small luteal cells were present in all corpora lutea, and were two to three times as numerous as large luteal cells. The former were irregular in shape, with tapering cytoplasmic processes. Their major cytoplasmic organelles were a predominantly smooth endoplasmic reticulum, mitochondria with tubular and lamellar cristae, and one or more Golgi complexes. The enzyme delta 5-3 beta-hydroxysteroid dehydrogenase was present in their cytoplasm. Small luteal cells were often interposed between large luteal cells and capillaries, and formed close, complex surface relationships with large luteal cells. Small and large luteal cells differed in many ways, including the restriction of numerous approximately 0.2 micron cytoplasmic granules to the large cells, and no cells of intermediate structure were observed. These features of small luteal cells suggest a steroid hormone synthetic function, and direct interaction with large luteal cells.


Observations on the mode of termination and parenchymal distribution of the splenic artery with evidence of splenic lobation and segmentation.

The mode of termination of the splenic artery was studied in 25 cadavers. Observation on the parenchymal distribution of the artery in 17 casts revealed avascular planes that divided the spleen into lobes, inside which other avascular planes separated the lobes into segments.


Experimental degeneration of intra-epithelia nerve fibres in cat airways.

This paper describes a quantitative and ultrastructural study of the degeneration of the intra-epithelial nerves in the epithelium of the lower respiratory tract of the cat following unilateral cervical infra-nodose vagotomy and section of the superior laryngeal nerve. A significant reduction in the number of intra-epithelial axons was found on the denervated side, degeneration being more complete at the hilus than in the trachea. Cellular inclusions resembling degenerating axons were observed especially on the denervated side, and their ultrastructural morphology is described. The functions of nerves which degenerated, and of those which remained unaffected, are discussed.


Cytochemical studies of cell population in peripheral blood smears of two West African teleosts.

The peripheral blood cells of two West African teleosts were examined for lipids, glycogen, desoxyribonucleic acid, succinic, lactic and glucose-6-phosphate dehydrogenases as well as DPNH- and TPNH-diaphorases, using cytochemical staining techniques. Our results suggest that both erythrocytes and thrombocytes as well as leucocytes are highly active metabolically, as indicated by strong reactions for LDH, glucose-6-phosphate dehydrogenase, DPNH- and TPNH-diaphorases. There is no evidence for the Krebs' cycle in any of the blood cells, The nuclei of erythrocytes and thrombocytes appear to be sites of activity of 'mitochondrial' type.


Neutron diffraction studies of lumbar vertebrae.

Neutron diffraction measurements of the orientation of the apatite crystals show a significant preference of their c-axes for the vertical direction. This preference increases from bottom to top of each vertebral body and is substantially greater for the second than for the fifth lumbar vertebrae. This is in line with the predominantly vertical forces which the former withstands.


An ultrastructural study of the endonasal microcirculation in the Wistar rat during fetal and early postnatal life.

Fenestrated capillaries were observed prenatally in the lateral nasal wall and in glandular capillaries of the nasal mucosa of the rat. The number of fenestrations increased towards full term and during postnatal life. Specific ultrastructural features in the vascular bed were found to be primarily determined in utero.


Diaphysial nutrient foramina in human phalanges.

1770 phalanges of the hand and 660 phalanges of the foot, of unknown sex, were studied to determine the number, position and direction of the nutrient foramina. It was found that, in general, these bones had one nutrient foramen in the middle third of the shaft. The proximal phalanges of the fingers often showed double foramina. Most of the foramina were situated on the border between the palmar/plantar and dorsal surfaces. The distal phalanges of the toes, and particularly of the great toe, may have a nutrient foramen on the dorsal surface. In the intermediate phalanges of the toes it is not possible to identify the nutrient foramen with certainty. All the foramina were directed away from the growing end.


Changes in axonal numbers in developing human trochlear nerve.

Complete axonal counts have been made in the intracranial parts of trochlear nerves from human fetuses of 9.2, 10 and 24 cm crown-rump length. A count was also made in the intraorbital part of the nerve from the 10 cm specimen. Schwann cell nuclei were also counted in typical cross sections, but do not necessarily reflect very accurately the schwann cell contents of the nerves. Axonal numbers conform to the propositions (1) that they do not all grow out at once, (2) do not all survive and (3) that degeneration may occur before or after myelination has begun. It seems inevitable that some loss of Schwann cells occurs in relation to the degeneration of myelinated axons, but there is no evidence for or against such a loss in relation to the degeneration of unmyelinated axons. Overall, however, Schwann cell numbers tend to increase as the number of myelinated axons increases.


An analysis of implantation in Indian hipposiderid bats.

The meagre information on the early developmental stages and the evolutionary and taxonomic position of hipposiderid bats has stimulated the present study. The cranial segment of the uterus forms a balloon-like enlargement at the time of implantation, and the antimesometrial part of its huminal slit expands into a spacious implantation chamber. Implantation is superficial and circumferential and the embryonic disc is orientated towards the side of the uterus which lies between the lateral and the antimesometrial sides. Reichert's membrane develops early, and the formation of a symplasma is described. A comparative account of implantation in the four species of hipposiderid bats of the family Hipposideridae is given, and the taxonomic position of this family is discussed on the basis of the present observations and earlier work on the embryology of Chiroptera.


Innervation of the sinu-atrial node and neighbouring regions in two human embryos.

In human embryos of 20 to 23 mm (36 to 40 days) it is possible to identify on the right side a nerve that we may call the sinusal, which originates by several roots from the nervus vagus dexter (Figs. 1A, B, D), descending through the right ventrolateral face of the primary trachea and right bronchus (Fig. 2, arrows). Beaded in appearance, it gives a fine anastomotic branch which, passing in front of the arteria pulmonalis dextra, passes to the left side (Figs. 2B, C, D; AN). At this level it gives the large branch for the nodus sinoatrialis which, penetrating through the wall of the superior vena cava, provides a rich innervation for the nodus sinoatrialis which is already in an advanced stage of differentiation (Fig. 3, 2; Cy, D, AN). Afterwards it gives fine branches which, following the atrial fold, are distributed throughout the posterior face of the atrium dextrum (Fig. 3). It increases in diameter and, passing through the angle formed by the right pulmonary veins with the atrium dextrum, reaches the intrapericardial portion of the inferior vena cava in the vicinity of its outlet from the atrium (Fig. 3, arrows). The whole innervation is parasympathetic at the stages studied.


The fine structure of the proximal growth plate of the avian tibia.

The ultrastructure of the proximal tibial growth plate of the 7 weeks old chicken has been described. Little ultrastructural difference could be ascertained between growth plates examined from normal white leghorn and commercial broiler chickens. The growth plate may be divided into five zones: interstitial, proliferating, prehypertrophic, hypertrophic, and degenerating hypertrophic. These zones reflect a maturation of chondrocytes, beginning with a stage of high mitotic and cytoplasmic activity passing through a stage of active secretion of matrical components (prehypertrophic and hypertrophic) and ending with degeneration of the cells and calcification of the matrix. Mineralization of the matrix appears to be initiated within matrical dense bodies, as in the mammal. Single hydroxyapatite crystals are first encountered about 0.1 mm proximal (i.e. towards the knee) to the limit of metaphyseal blood vessel ingrowth, while dense calcification is observed 0.1 mm distal to the tips of these metaphyseal vessels. The diameter of microfibrillary collagen in the growth plate matrix ranges from approximately 9 nm in the proximal zones to 19 nm in the distal zones. Many of the fibrils in the distal zones have a more or less distinct periodicity. Other major elements of the growth plate matrix are the ruthenium red-stained syncytial aggregates of mucopolysaccharides which are probably derived from the granules within the large intracellular Golgi vesicles. These findings have led the author to conclude that, while light microscopy indicates that avian and mammalian growth plates have very different structures, electron microscopy finds many similarities, suggesting that the physiological control mechanisms in these two vertebrate classes have much in common.


The uptake of horseradish peroxidase by damaged autonomic nerves in vitro.

The uptake of horseradish peroxidase by damaged autonomic nerves was studied in vitro. Large amounts of the enzyme penetrated the axons in a diffuse (nonvesicular) form. This occurred within 90 minutes of application of the enzyme. Twenty four hours after application of the enzyme diffuse peroxidase was still present in the axoplasm, but it was 'diluted' as a result of great axonal swelling. Pinocytotic uptake of the enzyme was observed both 90 minutes and 24 hours after application of the enzyme. The swollen axons close to the point of ligation showed most evidence of pinocytotic uptake, largely into coated vesicles, and much membrane-bound peroxidase reaction product was present. The uptake of peroxidase in a membrane-bound form may be of particular significance for the subsequent retrograde transport of the enzyme.


New antibiotics from Actinoplanes strains. Structure of A 17002 C.

A 17002 C is a new metabolite produced by Actinoplanes strains, structurally related to the virginiamycin factor M. On the basis of physico-chemical data, MS, IR, 1H and 13C NMR, structure I is assigned to A 17002 C.


Isolation of a new peptide antibiotic, permetin A, from Bacillus circulans.

Permetin A was purified from the culture filtrate of Bacillus circulans AJ 3902 by extraction with n-butanol, precipitation with sodium helianthate, CM-cellulose column chromatography and Sephadex LH-20 column chromatography. The compound was found to be a new peptide antibiotic containing 2,4-diaminobutyric acid (Dab), leucine, isoleucine, phenylalanine, valine, serine (in a molar ratio of 3:2:1:1:1:1) and a fatty acid. This antibiotic showed activity in vitro against Gram-negative, Gram-positive and some anaerobic bacteria.


The structure of permetin A, a new polypeptin type antibiotic produced by Bacillus circulans.

The structure of permetin A(I), an antibiotic substance produced by Bacillus circulans AJ 3902, has been elucidated as a cyclic acyl peptide by means of the mass and nuclear magnetic resonance spectroscopic techniques. (Formula: see text.) The structure was found to be the same as polypeptin A(II) except that L-Thr in II is replaced by L-Ser in I. Details of the structural determination are given for the permetin A itself as well as for the hydrolyzed permetin A. (Formula: see text.)


Early infantile autism in monozygotic twins.

A pair of male monozygotic twins concordant for autism is reported. During pregnancy the mother suffered from severe toxemia, and delivery occurred 2 months before term. Although there may have been a genetic influence, it appears that gestational damage was the main etiological factor for the autism in both children.


Effects of peer-mediated social initiations and prompting/reinforcement procedures on the social behavior of autistic children.

Peer-mediated social initiations and prompting/reinforcement procedures were evaluated (in training and generalization sessions) as interventions for increasing the positive social behavior of four autistic children. During baseline, the peer trainer made few social initiations and did not prompt or socially reinforce subjects. For two subjects, baseline was followed by social initiation intervention, and for the other two, baseline was followed by prompting and social reinforcement. Both interventions produced dramatic and comparable increases in positive social behavior in training sessions. Post-treatment responding was not observed for either intervention. When interventions terminated in a second baseline period, the subjects' behavior returned to the level observed during the initial baseline. The subjects were then exposed to the intervention procedure they had not yet experienced. Again, there were positive and comparable behavior changes in the treatment setting, but no increase in positive social behavior was observed during generalization assessment.


Histidinemia and infantile autism.

A 10-year-old boy first showed features of infantile autism at age 24 months. Histidinemia was also diagnosed, with histidine blood levels seven times higher than the upper normal values. If the coexistence of autism and histidinemia was not coincidental, histidinemia may have constituted a necessary but not sufficient factor leading to the clinical condition of autism. Other members of the patient's family had high blood levels of histidine, but did not show symptoms that have been related to histidinemia.


Free-play behavior of atypical children: an approach to assessment.

Children's free-play behavior and its relationship to IQ and usefulness in assessing severely disturbed children were investigated. The results indicated that (1) the play of 100 normal children scored for quality and time generally yielded significant positive correlations with Binet and WPPSI IQs; (2) the correlations remained the same when verbalizations were deleted from the play protocols; (3) the play scores for 16 severely disturbed boys significantly differentiated two subgroups differing in degree of pathology, showed significant improvement from the pre- to posttreatment periods, and indicated pretreatment scores were significantly and positively correlated with improvement assessed by observer ratings in a 2-year follow-up. Implications for the use of this measure in assessment and the role of play in development were discussed.


Plasma levels of folates, riboflavin, vitamin B6, and ascorbate in severely disturbed children.

The recent upsurge in megavitamin therapy raises questions about the role of vitamin deficiencies and dependencies in mental health. With this in mind, the plasma levels of folic acid, ascorbic acid, pyridoxine, and riboflavin were studied in approximately 125 children admitted to a child psychiatric unit. There were no apparent decreased levels of vitamins in these children in terms of their age, race, or psychiatric diagnosis. It is postulated that vitamin deficiencies per se cannot be proposed as etiological factors in any of the psychiatric deficits represented. Megavitamin therapy, if successful, is not effective due to crrection of vitamin deficiencies as opposed to vitamin dependencies and may be due to the metabolic onus and consequent effects of such heavy doses of vitamins.


Effects of age on communication skill levels and prevalence of maladaptive behaviors in autistic and mentally retarded children.

This study investigated the effect of age on communication and maladaptive behavior in autistic and mentally retarded children. Forty-seven autistic and 128 mentally retarded children in a special school served as subjects. The cross-sectional method was used to compare junior and senior groups, and ratings on communication skills and maladaptive behaviors were obtained from teachers. We found that the skills of comprehension and conversation in autistic children improved significantly with age, and speech improved somewhat. In spite of this improvement in communication skills, maladaptive behaviors in the autistic children other than hyperactivity did not change significantly with age. Withdrawal improved significantly with age in the mentally retarded children but not in the autistic children.


Relationship between lysostaphin endopeptidase production and cell wall composition in Staphylococcus staphylolyticus.

Mutants of Staphylococcus staphylolyticus incapable of producing an extracellular staphylolytic glycylglycine endopeptidase were isolated and found to have cells in the population susceptible to lysis by this enzyme, as did the wild-type organism under conditions in which the endopeptidase was not produced. These results suggest that cultures of this organism normally contain a heterogeneous population of cells with regard to cell wall composition and susceptibility to the enzyme. Production of the endopeptidase appears to act as a selective pressure which removes the susceptible cells in the population as the enzyme appears in the medium. A comparison of the peptidoglycan of the wild-type organism grown under conditions in which the endopeptidase was produced with that of this organism grown under nonproducing conditions and with those of endopeptidase-less mutants showed that in the presence of the endopeptidase the cell population had peptidoglycan with shorter peptide cross bridges and a greater percentage of serine in these cross bridges than was found in cells grown in the absence of the enzyme. The inability of the endopeptidase to hydrolyze glycylserine and serylglycine peptide bonds suggests that at least part of the resistance this organism has to the endopeptidase is due to relative amounts of serine found in the peptide cross bridges of some cells in the population.


Release of lipoteichoic acid from Streptococcus sanguis: stimulation of release during penicillin treatment.

The spontaneous and the penicillin-stimulated release of water-soluble, glycerol-labeled polymers was compared in Streptococcus sanguis. In contrast to the spontaneous release occurring in exponentially growing or stationary-phase bacteria, penicillin-treated cells released the bulk of these polymers, and they were not replenished by synthesis during antibiotic treatment. Furthermore, a major portion of the extracellular polymers was characterized as acylated lipoteichoic acid.


Growth yields and fermentation balance of Bacteroides fragilis cultured in glucose-enriched medium.

Bacteroides fragilis is an obligate anaerobic bacterium classified with the gram-negative, non-sporeforming bacilli and is the Bacteroides species most frequently isolated from human infections. In the present study, experiments were designed to investigate growth characteristics of B. fragilis in a complex medium. In a minimal defined medium, which was employed for comparison purposes, B. fragilis grew with a generation time of 2 h. Growth of the organism in glucose-enriched medium used in the present study was superior. Maximum generation time was 60 min. Total and viable cells (colony-forming units) were 8.9 x 10(9) and 2.1 x 10(9), respectively, at maximum measurable growth. The molar growth yield (Ym) was 51.5. Growth yields were found to reach a maximum 2 to 3 h before maximum growth and to vary with respect to the phase of growth. Estimates of the fermentation products indicated that glucose was the sole energy substrate. Major products included acetic acid, propionic acid, lactic acid, and succinic acid. Other products included ethyl alcohol, pyruvic acid, and fumaric acid. No attempt was made to recover CO2 or formic acid. The OR balances from two experiments were 0.013 and -0.093 and the respective carbon recoveries were 6.268 and 6.241. The results of the present study show that B. fragilis is capable of rapid rates of growth in vitro by using glucose as the sole energy source.


Isolation and characterization of protoplasts from Saccharomyces rouxii.

Cells of the osmotolerant yeast Saccharomyces rouxii were transformed to protoplasts in good yield (85%) by digesting cell walls with snail-gut enzyme in the presence of 10 mM dithioerythritol, 0.1 M sodium phosphate buffer (pH 6.8), and 2.0 M KCl. The requirement for 2.0 M KCl compares with that for S. bisporus var. mellis (another osmotolerant species) and contrasts with the 0.3 to 0.8 M KCl concentrations used in the preparation of most yeast protoplasts. Short digestions (60 min or less) produced mostly spheroplasts; longer incubations (90 min or more) yielded mostly protoplasts as judged by electron micrographs. These protoplasts could be transferred to 1.0 M KCl or 2.0 M sorbitol without lysing, but lysis was pronounced in 0.5 M KCl or 1.0 M mannitol and complete in 0.02 M KCl. Protoplasts were separated from isolated cell wall remnants and debris by centrifugation on a linear gradient of Ficoll 400 (35 to 17.5%, wt/vol) containing 2.0 M KCl. Both crude and fractionated protoplast preparations contained vesicles which were identified with the periplasmic bodies of whole cells. Some of the periplasmic bodies were connected to protoplasts by fine pedicels; others appeared free. Independent degeneracy of periplasmic bodies was occasionally observed. beta-Fructofuranosidase (EC 3.2.1.26) activity is cryptic (physically) in cells of S. rouxii in contrast to the expressed enzyme (periplasmic space) of other Saccharomyces species. This enzyme remains cryptic in protoplast preparations of S. rouxii but is expressed upon lysis. The same specific activities were found per unit cell or protoplast. The possible association of the cryptic enzyme with periplasmic bodies is discussed.


Axial filament involvement in the motility of Leptospira interrogans.

Motility mutants of Leptospira interrogans serovar illini were isolated and analyzed by dark-field and electron microscopy. Mutants were obtained by screening for small colonies after nitrosoguanidine treatment. One class of mutants did not have hook- or spiral-shaped ends. In addition, the axial filaments from these mutants were not coiled. An analysis of revertants of two of the mutants in this class indicated that the mutations were pleiotropic with respect to motility, hook- and spiral-shaped ends, and axial filament coiling. We conclude that the axial filaments and the hook- and spiral-shaped ends are involved in L. interrogans motility.


Helix handedness of Leptospira interrogans as determined by scanning electron microscopy.

Representative serovars and strains of the seven genetic groups of Leptospira interrogans, and two previously studied serovars, were all found to form exclusively right-handed helices as determined by scanning electron microscopy. No change in handedness occurred in cells grown in a minimal medium (Tween-80 albumin) compared to cells grown in a rich medium (rabbit serum). The right-handedness of the organisms was related to the evolution, cell wall structure, and the mechanism of motility of L. interrogans.


Association of flexing and gliding in Flexibacter.

Nongliding mutants of Flexibacter FS-1 are unable to flex; revertants regain both forms of movement. A variety of conditions and treatments reversibly inhibit both gliding and flexing.


Deoxyribonucleic acid-mediated transformation of Spirillum lipoferum.

The methodology for deoxyribonucleic acid-mediated transformation of Spirillum lipoferum to resistance to various antimicrobial agents is reported.


Fatty acids in phospholipids of cells, cysts, and germinating cysts of Azotobacter vinelandii.

Cyclopropane fatty acids constitute 25% of the phospholipid acyl groups in cysts of Azotobacter vinelandii. These are lost by dilution during germination when the synthesis of the fatty acids characteristic of vegetative cell phospholipids commences.


23S ribosomal ribonucleic acid of macrolide-producing streptomycetes contains methylated adenine.

Coresistance to macrolide, lincosamide, and streptogramin B-type (MLS) antibiotics by a common biochemical mechanism characterizes clinically resistant pathogens. Of 10 streptomycetes tested for resistance to macrolide, lincosamide, and streptogramin B-type antibiotics, only 1, Streptomyces erythreus, the organism used for production of erythromycin, was found resistant to all three classes; moreover, it was the only streptomycete in the series tested found to contain N6-dimethyladenine (m62A) in 23S ribosomal ribonucleic acid, the structural alteration of ribosomal ribonucleic acid associated with clinical resistance. Of the seven streptomycetes tested for the presence of m62A and N6-methyladenine (m6A), two, S. fradiae and S. cirratus, which produce the macrolide antibiotics tylosin and cirramycin, respectively, were found to contain m6A, but not m62A. The remaining strains tested, including strains which produce lincomycin and streptogramins, contained neither m6A nor m62A.


Suppression of polar effects of nonsense mutations by ultraviolet irradiation.

Nonsense mutations in capsid genes F and G of phages S13 and phi X174 decreased the expression of genes downstream. These polar effects were suppressed by ultraviolet irradiation of the host before infection. Activities of the downstream genes were restored to between 30 and 95% of their normal levels, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the viral proteins. The effect of ultraviolet irradiation may be to suppress rhodependent termination of transcription. The polarity of one mutation was poorly suppressed by ultraviolet irradiation.


Location of peptidoglycan lytic enzymes in Bacillus sphaericus.

The level of three peptidoglycan hydrolases was determined in the mother cell compartment and forespores of  Bacillus sphaericus. Vegetative and sporulating cells contained in LD-carboxypeptidase active only on the vegetative cell wall peptidoglycan, and we have previously shown that sporulation is accompanied by the production of two new enzymes active only on the spore cortex peptidoglycan. These gamma-D-glutamyl-meso-diaminopimelate endopeptidase and a meso-diaminopimelate-D-alanine dipeptidase. The LD-carboxypeptidase activity appeared to be located in the membranes of both the mother cells and forespores. Endopeptidase activity was located in the integument fraction of the forespores, and the dipeptidase activity was only found in the forespore cytoplasm. These different locations comply with the probable different functions of these enzymes.


Superoxide dismutase and O2 lethality in Bacteroides fragilis.

Exposure of midlog Bacteroides fragils (VPI 2393) to 2% O2-98% N2 caused a three- to fivefold increase in superoxide dismutase specific activity within the cells. The increase in specific activity was completed within 90 min after exposure to oxygen and was dependent upon protein synthesis. Cells containing the higher superoxide dismutase level were more resistant to the effects of 5 atm of oxygen tension than were cells containing the lower level of superoxide dismutase but were equally resistant to 5 atm of nitrogen tension. Similar results were observed upon comparing viability experiments with B. fragilis and B. vulgatus. Superoxide dismutase activity in sonic extracts of B. fragilis was rapidly inactivated by exposure to 5 mM H2O2 and was inhibited by 1 mM NaN3 but not 5 mM NaCN. The inhibition pattern is identical to the pattern demonstrated for the purified iron-containing enzyme from Escherichia coli B and suggests that the superoxide dismutase in B. fragilis is an iron enzyme.


Modifiable chromatophore proteins in photosynthetic bacteria.

The chromatophores of Chromatium vinosum, as well as six other photosynthetic bacteria, contained two or more proteins which were insoluble when heated in the presence of sodium dodecyl sulfate (SDS) and 2-mercaptoethanol (beta-ME). When the chromatophores were dissolved at room temperature in SDS-beta-ME, these proteins were present in the SDS-polyacrylamide gel electrophoresis profiles, but when the samples were dissolved at 100 degrees C, they were absent or considerably diminished. When one-dimensional gels of chromatophores solubilized at room temperature were soaked in the SDS-beta-ME solution and heated to 100 degrees C and the gels were run in a second dimension, the proteins became immobilized in the original first-dimension gel, where they could be detected by staining. The two major proteins so affected in C. vinosum had apparent molecular weights of 28,000 and 21,000. The chromatophores of several other photosynthetic bacteria also contained predominant proteins between 30,000 and 19,000 molecular weight, which became insoluble when heated in the presence of SDS and beta-ME. In at least two of the species examined, these appeared to be reaction center proteins. The conditions causing the proteins to become insoluble were complex and involved temperature, SDS concentration, and the presence of sulfhydryl reagents. The chromatophores of four of the Chromatiaceae species and two strains of one of the Rhodospirillaceae species examined had a protein-pigment complex that was visible in SDS-polyacrylamide gel profiles of samples dissolved at room temperature but was absent in samples dissolved at 100 degrees C.


Arrays of hemispheric surface projections on Chlamydia psittaci and Chlamydia trachomatis observed by scanning electron microscopy.

Scanning microscopy of two strains of Chlamydia psittaci and four strains of Chlamydia trachomatis representative of the wide diversity in origin and behavior of members of the genus revealed patches of regular arrays of hemispheric projections on the surfaces of elementary bodies of all six strains. These distinctive and perhaps unique surface structure are probably present in all populations of chlamydiae.


Bidirectional replication of the mini-ColE1 plasmid pVH51.

Replicating molecules of the min-ColE1 plasmid pVH51 have been examined by electron microscopy after cleavage with the restriction endonuclease EcoRI. Replication apparently starts at a unique site indistinguishable from the origin of replication used by the parental plasmid ColE1. In contrast to ColE1, the structure of the majority of the replication intermediates was consistent with a bidirectional mode of replication. A minor portion of the molecules appeared to replicate unidirectionally in either direction from the same origin.


Isolation and characterization of a cycloheximide-resistant mutant of Acanthamoeba castellanii Neff.

A cycloheximide-resistant mutant was isolated from the amoeba Acanthamoeba castellanii Neff. Drug resistance was found to be due to a ribosomal modification.


Fatty acid composition of lipopolysaccharides of Vibrio cholerae 35A3 (Inaba), NIB 90 (Ogawa), and 4715 (Nag).

Considerable amounts of odd-numbered fatty acids, such as non-hydroxy C15 and C17 and 3-hydroxy C11 and C13 acids, were found in lipopolysaccharides from Vibrio cholerae 35A3 (Inaba).


Induction and catabolite repression of L-rhamnose dehydrogenase in Pullularia pullulans.

The growth of Pullularia pullulans on L-rhamnose (6-deoxy-L-mannose) as the sole carbon source induces the synthesis of L-rhamnose dehydrogenase, a nicotinamide adenine dinucleotide-dependent enzyme that catalyzes the oxidation of the deoxy sugar to L-rhamnonolactone. The enzyme induction is inhibited by cycloheximide, suggesting de novo synthesis. The presence of d-glucose (0.2%) or D-galactose (0.2%) simultaneously with the inducer in the induction medium produced 50% repression of dehydrogenase synthesis, but no effect was detected with D-fructose and D-mannose at the same concentration. High levels of D-glucose (2%), under maximal catabolite repression conditions, produced a complete inhibition of enzyme synthesis.


Amino acid catabolism and malic enzyme in differentiating Dictyostelium discoideum.

Amino acids produced from protein degradation are the major energy source for differentiation and aging in Dictyostelium discoideum. Considering the reactions involved in the conversion of amino acids from an average protein into tricarboxylic acid cycle intermediates, a route from a cycle intermediate (probably malate) to acetyl coenzyme A is required for the complete utilization of amino acids. Citrate was isolated from cells pulse-labeled with (14)C-labeled amino acids and was cleaved with citrate lyase. When cells were pulse-labeled with [U-(14)C]-glutamate the specific radioactivity of the acetate and oxaloacetate portions of citrate were consistent with the conclusion that one-third of the carbon flowing through the tricarboxylic acid cycle is removed for the synthesis of acetyl coenzyme A. The data were also consistent with the patterns of carbon flux required to maintain steady-state levels of cycle intermediates in cells catabolizing amino acids. It is suggested that the malic enzyme (EC 1.1.1.40) catalyzes the synthesis of acetyl coenzyme A from malate and is responsible for the observed citrate labeling pattern. In cell extracts the activity of this enzyme increased markedly with the onset of differentiation. The properties of partially purified (40-fold) malic enzyme isolated at culmination indicated that the enzyme was allosteric and was positively affected by aspartate and glutamate. Thus, amino acid production from protein degradation would stimulate a reaction essential for the efficient utilization of these amino acids for energy.


Temperature-sensitive mutants of Physarum polycephalum: viability, growth, and nuclear replication.

Using a selfing strain of Physarum polycephalum that forms haploid plasmodia, we have isolated temperature-sensitive growth mutants in two ways. The negative selectant, netropsin, was used to enrich for temperature-sensitive mutants among a population of mutagenized amoebae, and, separately, a nonselective screening method was used to isolate plasmodial temperature-sensitive mutants among clonal plasmodia derived from mutagenized amoebae. Complementation in heterokaryons was used to sort the mutants into nine functional groups. When transferred to the restrictive temperature, two mutants immediately lysed, whereas the remainder slowed or stopped growing. Of the two lytic mutants, one affected both amoebae and plasmodia, and the other affected plasmodia alone. The growth-defective mutants were examined for protein and deoxyribonucleic acid synthesis and for aberrations in mitotic behavior. One mutant may be defective in both protein and deoxyribonucleic acid synthesis, and another only in deoxyribonucleic acid synthesis. The latter shows a striking reduction in the frequency of postmitotic reconstruction nuclei at the restrictive temperature. We believe that this mutant, MA67, is affected in a step in the nuclear replication cycle occurring late in G2. Execution of this step is necessary for both mitosis and chromosome replication.


Respiration in the yeast and mycelial phases of Histoplasma capsulatum.

Respiration in the yeast and mycelial phases of Histoplasma capsulatum proceeds via a cytochrome system and an alternate oxidase, both present constitutively. The mycelial cytochrome system is distinguished by an additional partial shunt around the antimycin-sensitive site.


Presence of a major (storage?) protein in dormant spores of the fungus Botryodiplodia theobromae.

Approximately 23% of the protein isolated from dormant spores of Botryodiplodia theobromae consisted of a single polypeptide; the polypeptide is probably degraded during germination.


The difficult patient: identification and response.

A group of "difficult" psychiatric outpatients in a university hospital system was identified and its distinguishing characteristics compared to a sample of other psychiatric outpatients not so labeled. "Difficult" patients were perceived to be significantly more demanding (p less than or equal to 0.005), dangerous, difficult to empathize with, manipulative and likely to polarize the staff. These patient were perceived differently by physician  and nonphysican staff. An analysis revealed that a major source of "difficulty" appeared to be the structure of the treatment system rather than the patient. A corrective strategy was devised to test this hypothesis.


Electroencephalographic abnormalities in suicidal patients.

Struve has suggested that paroxysmal EEG abnormality is considerably more frequent in suicidal psychiatric inpatients than in nonsuicidal psychiatric inpatients. In order to see if these observations were confirmed in other populations, the Zung Index of Potential Suicide and scalp EEG were administered to 216 consecutively admitted VA psychiatric inpatients. The results indicated only slightly greater EEG abnormality in suicidal patients (14.9%), than in nonsuicidal patients (6.6%) statistical analysis failed to support either a general association between overall EEG abnormality and suicidal behavior, or the specific association between paroxysmal EEG abnormality and suicidal behavior, reported by Struve. Some of the data suggested that further EEG observations might be worthwhile on patients in the Threatener subcategory of suicidal behavior.


Wechsler variables as a function of age and neurologic status.

Scores on Wechsler variables were analyzed in groups of patients broken down by age and neurologic status. No differences as a function of age were found in intelligence or memory. Timed Performance subtest results were related to age, but apparent Verbal subtest age differences were eliminated by the introduction of education as a covariate. Neurologic status showed significant effects on intelligence and memory and on all but 3 Wechsler subtests. No significant interactions between age and neurologic status were seen.


Evaluation of day treatment center effectiveness.

A study was undertaken to evaluate the efficacy of the Day Treatment Center as an alternative to rehospitalization for chronic psychiatric patients. The project utilized a subject as his own control design comparing equal time intervals before and during Day Treatment Center involvement. The study demonstrates the efficacy of the Day Treatment Center as a therapeutic modality that not only helps prevent hospitalization, but also assists in decreasing the average length of stay when admission is unavoidable.


Nocturnal levels of growth hormone in hyperactive children of small stature.

Growth disturbances have been demonstrated in hyperactive children treated with stimulant medication. This study examines sleep related growth hormone secretion in patients with hyperactive syndrome and relative short stature. Five such patients were compared with 9 age matched controls. There were no differences in sleep patterns, and growth hormone levels for the hyperactive children were within normal limits. This study suggests that hyperactive children with short stature have normal sleep patterns and normal sleep related growth hormone secretion.


The uses of seclusion on a general hospital psychiatric unit.

Seclusion was used in the management of 36.6% of the patients on a general hospital psychiatric unit during a 6 month prospective study. It was initiated most frequently to decrease stimulation for agitated patients with poor impulse control, between 10 pm and 2 am, and when the unit census was high. Patients who required seclusion were significantly younger, hospitalized longer, more often manic and received pharmacotherapy more frequently. No correlation was found between nursing staff age and psychiatric experience and the frequency that they initiated seclusion. The value of seclusion during a medication-free diagnostic assessment period is discussed.


Repertory grid and biochemical characterization of depression: a comparison of case reports.

Two cases of depression are presented: One which responded to psychotherapy and the other which responded to medication. These cases were evaluated by repertory grid analysis and by biochemical analysis. These results serve as the basis for a discussion of the use of such analysis as predictors of response in depression to psychotherapy or medication.


Delirium associated with combined fluphenazine-clonidine therapy.

The authors report a case of acute organic brain syndrome in a patient being treated with clonidine and fluphenazine that cleared when clonidine was discontinued. Theoretical considerations of dopamine-norephinephrine interactions are discussed in the context of the drug-drug interaction.


Immunoglobulin G antibodies to herpes simplex type 1 virus detected by radioimmunoassay in serum and cerebrospinal fluid of patients with schizophrenia.

Serum and CSF specimens from 16 schizophrenic patients and 18 nonpsychiatric controls were tested by radioimmunoassay for immunoglobulin G antibody of capsid, envelope and excreted antigens of herpes simplex type 1 virus. There were no significant differences in the antibody levels between the schizophrenic patients and the controls. The etiological role of viruses and virus-like agents in schizophrenia and some methodological aspects are discussed.


Structural characteristics of interferons from mouse Ehrlich ascites tumor cells.

An improved procedure for the isolation of interferons produced by mouse Ehrlich ascites tumor cells infected with Newcastle disease virus provides interferons of three size classes (33,000, 26,000, and 20,000 daltons) with specific activities between 2 and 3 x 10(9) units/mg of protein and a yield of 11 to 20%. The tryptic peptide maps of the two larger species are very similar; that of the smallest species is different, at least in part. The amino acid compositions of the three species are very close. Their NH2-terminal amino acids are identical and so are the amino acids released by carboxypeptidase A treatment. These data are consistent with the possibility that the differences in size between the three species may be due, at least in part, to unequal glycosylation.


Identification of in vitro synthesized pituitary glycoprotein alpha subunit. Translation of a possible precursor.

Bovine pituitary RNA was translated in heterologous cell-free systems derived from wheat germ and reticulocyte lysate. Analyses of the cell-free products by sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed three major proteins, exhibiting apparent molecular weights of 25,000, 24,000, and 14,000. The two larger products were identified as preprolactin and pregrowth hormone by immunoprecipitation and thus demonstrated the fidelity of pituitary RNA translation. The 14,000-dalton product was shown to be immuno-precipitable with specific bovine lutropin (LH)alpha antisera. Since this protein is 3000 to 4000 daltons larger than the apoprotein form of the alpha subunits, it suggests that the subunit is synthesized in precursor form. The immunological specificity was further demonstrated by the successful competition with unlabeled alpha subunit plus the failure to immunoprecipitate this product using specific antisera to other pituitary hormones. Although specific antisera to bTSH(thyrotropin)beta and bLH(lutropin)beta failed to immunoprecipitate the 14,000-dalton product, LHbeta antisera precipitated a product with a molecular weight of approximately 18,000. Since the alpha and beta antisera specifically precipitated different products, and since a larger immunoprecipitable product was not detected, the results suggest that the two subunits are synthesized separately.


Calcium-dependent activation of a multifunctional protein kinase by membrane phospholipids.

The proenzyme of a Ca2+-dependent protease-activated protein kinase previously obtained from mammalian tissues (Inoue, M., Kishimoto, A., Takai, Y., and Nishizuka, Y. (1977) J. Biol. Chem. 252, 7610-7616) was enzymatically fully active without limited proteolysis when Ca2+ and a membrane-associated factor were simultaneously present in the reaction mixture. The activation process was reversed by removing Ca2+ with ethylene glycol bis(beta-aminoethyl ether)N,N,N',N'-tetraacetic acid. An apparent Ka value for Ca2+ was less than 5 x 10(-5) M. Other divalent cations were inactive except for Sr2+, which was 5% as active as Ca2+. The factor was almost exclusively localized in membrane fractions of various tissues including brain, liver, kidney, skeletal muscle, blood cells, and adipose tissue. It was easily extractable with chloroform/methanol (2:1), and was recovered in the phospholipid fraction. In fact, this membrane factor could be replaced by chromatographically pure phosphatidylinositol, phosphatidylserine, phosphatidic acid, or diphosphatidylglycerol. Phosphatidylethanolamine, phosphatidylcholine, and sphingomyelin were far less effective under the comparable conditions. Ca2+-dependent modulator protein was unable to support enzymatic activity. The enzyme thus activated showed an ability to phosphorylate five histone fractions and muscle phosphorylase kinase, and appeared to possess multifunctional catalytic activities.


Developmental change and genetic defect in the carbohydrate structure of band 3 glycoprotein of human erythrocyte membrane.

The chemical structure of Band 3 glycopeptide prepared from erythrocytes of normal adult (blood group OI), umbilical cord vessels (Oi), and an i adult variant who fails to develop I antigen (Oi), has been compared. Band 3 glycopeptide of cord erythrocytes gave, on permethylation analysis, predominantly 2,4,6-tri-O-methylgalactose and 3,6-di-O-methyl-2-N-methylacetamido-2-deoxyglucose, whereas the same glycopeptide of normal adult erythrocytes gave much higher amounts of 2,3,4,6-tetra-O-methylgalactose and 2,4-di-O-methylgalactose as compared with that of cord erythrocytes. Band 3 glycopeptide from i adult showed the same methylation pattern as cord-Band 3 glycopeptide. In accordance with these results, Band 3 glycopeptide of cord and i adult erythrocytes were hydrolyzed to mostly small oligosaccharides by endo-beta-galactosidase from Escherichia freundii, whereas that of normal adult produced a number of oligosaccharides with various sizes which was caused by branched structures. Based on these results and structures of released oligosaccharides, the major developmental change of carbohydrate structure in the erythrocyte membrane is the conversion of linear repeating Galbeta1 leads to 4GlcNAcbeta1 leads to 3Gal to a branched Galbeta 1 leads to 4GlcNAcbeta 1 leads to 3 (R leads to 6) Gal structure. i individual may result from the lack of the branching enzyme.


Evidence that a single GTP is used in the formation of 80 S initiation complexes.

Evidence is presented that the GTP initially bound in ternary complex (Met-tRNAf.GTP.eukaryotic initiation factor 2 (eIF-2)) is the same GTP that is hydrolyzed to allow joining of a 40 S preinitiation complex with 60 S subunits. This evidence was obtained by two quite dissimilar techniques. The first was a kinetic analysis of AUG-directed methionyl-puromycin synthesis using either eIF-2 of eIF-2A to direct the binding of Met-tRNAf to 40 S subunits. The second technique was the isolation of 40 S preinitiation complexes by Sepharose 6B chromatography and subsequent quantitation of GTP hydrolysis and methionyl-puromycin synthesis under conditions where 80 S complex formation is permitted.


Demonstration of a rat liver microsomal binding protein specific for beta-glucuronidase.

A binding protein with apparent specificity for beta-glucuronidase has been partially purified from a Triton X-100 extract of rat liver microsomes by affinity chromatography on glucuronidase-Sepharose 2B. It appears that once removed from the membrane, this binding protein self-aggregates to form large macromolecular complexes. With the use of polyacrylamide gel electrophoretic and sucrose density gradient ultracentrifugation assays to monitor the conversion of glucuronidase tetramer to a very high molecular weight complex, it was shown that the binding activity is heatlabile and protease-sensitive. However, binding activity is not influenced by salts, carbohydrates, other proteins or glycoproteins, or by extensive periodate oxidation of beta-glucuronidase, nor does binding occur with any other protein tested. The binding protein does not discriminate against any form of beta-glucuronidase from any rat organ tested. However, the binding protein does show organ localization, being present in the liver and kidney but not the spleen. The possible relationship of this binding protein to egasyn, a membrane protein which stabilizes beta-glucuronidase in mouse liver endoplasmic reticulum, is discussed.


Protonmotive force and catecholamine transport in isolated chromaffin granules.

The effect of the transmembrane potential (delta psi) and the proton concentration gradient (delta pH) across the chromaffin granule membrane upon the rate and extent of catecholamine accumulation was studied in isolated bovine chromaffin granules. Freshly isolated chromaffin granules had an intragranular pH of 5.5 as measured by [14C]methylamine distribution. The addition of ATP to a suspension of granules resulted in the generation of a membrane potential, positive inside, as measured by [14C]thiocyanate (SCN-) distribution. The addition of carboxyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP), a proton translocator, resulted in a reversal of the potential to negative values (measured by [3H]tetramethylphenylphosphonium (TPMP+)) approaching -90 mV. Changing the external pH of a granular suspension incubated with FCCP produced a linear perturbation in the measured potential from positive to negative values, which can be explained by the distribution of protons according to their electrochemical gradient. When ammonia (1 to 50 mM) was added to highly buffered suspensions of chromaffin granules there was a dose-dependent decrease in the transmembrane proton gradient (delta pH) and an increase in the membrane potential (delta psi). On the other hand, thiocyanate or FCCP, at varying concentration, produced a dose-related collapse of the membrane potential and had no effect upon the transmembrane proton gradient. The addition of larger concentrations of catecholamines caused a decrease in the transmembrane proton gradient and an increase in the membrane potential. Time-resolved influx of catecholamines into the granules was studied radiochemically using low external catecholamine concentrations. The accumulation of epinephrine or norepinephrine was over one order of magnitude greater in the presence of ATP than in its absence. The rate and extent of amine accumulation was found to be related to the magnitude of the membrane potential at fixed transmembrane proton concentration (delta pH) values. Likewise, the accumulation was related to the magnitude of the delta pH at fixed membrane potential values. These results suggest that the existence of both a transmembrane proton gradient and a membrane potential are required for optimal catecholamine accumulation to occur.


Effect of polyanions on the kinetics of the reaction of apohemoglobin with carbonmonoxy heme.

The reaction of apohemoglobin with carbonmonoxy heme and with carbonmonoxy heme dimethyl ester was investigated in the presence and absence of inositol hexaphosphate. The binding stoichiometry of both heme derivatives to apohemoglobin was not affected by the presence of the polyphosphate, while, in both cases, the overall rate of recombination was substantially decreased. The absence of the negatively charged carboxyl groups in the dimethyl ester derivative of the heme indicated that the effect of inositol hexaphosphate on the reaction of apohemoglobin with heme was not due to electrostatic repulsions and resulted from conformational changes occurring upon the interaction of apohemoglobin with inositol hexaphosphate. Qualitative treatment of the kinetic data suggests that these conformational changes destabilize the intermediates of the reaction by increasing their redissociation into the original components. Also, benzenehexacarboxylate produced conformational changes in apohemoglobin and decreased its rate of reaction with carbonmonoxy heme, proving the aspecificity of the interaction of apohemoglobin with polyanions.


Biohydrogenation of unsaturated fatty acids. Presence of dithionite and an endogenous electron donor in Butyrivibrio fibrisolvens.

Two oxygen-consuming substances were isolated from cell-free extracts of the rumen anaerobe, Butyrivibrio fibrisolvens. The major fraction comprising 97% of the total activity was characterized as a three-component mixture of glucose, maltose, and dithionite. The minor activity fraction contained an electron donor for the reduction of cis-9,trans-11-octadecadienoate to trans-11-octadecenoate. After oxidation, the electron donor could be reduced by the dithionite, thereby accounting for the previously observed capacity of cell-free extracts of the bacterium to carry out the biohydrogenation of the conjugated dienoic fatty acid.


Conversion by rat brain preparation of lignoceric acid to ceramides and cerebrosides containing both alpha-hydroxy and nonhydroxy fatty acids. Effects of compounds which affect sphingolipid metabolism.

Three synthetic compounds which affect sphingolipid metabolism in vitro were examined for their effects on the synthesis in a rat brain system of nonhydroxyceramide, hydroxyceramide, nonhydroxycerebroside, and hydroxycerebroside from [1-14C]ignoceric acid. These compounds are N-octanoyl-D-threo-p-nitrophenylaminopropanediol (Compound I), N-(2,3-epoxydecanoyl)-norephedrine (Compound II), and N-decyl-N'-glucosylthiourea (Compound III). In the presence of up to 0.5 mM Compound I, only the hydroxyceramide fromation increased, while the synthesis of the other three lipids decreased. Compound II strongly inhibited the formation of all four lipids at all concentrations tested (0.1 to 1 mM). Compound III increased the synthesis of both ceramides approximately 2-fold at 1 mM, but the conversion of lignoceric acid to both cerebrosides remained relatively unchanged. Several significant conclusions from these observations are discussed.


Solubilization and characterization of a platelet membrane ADP-binding protein.

Previous studies have shown that platelet membranes bind radiolabeled ADP and have nucleoside diphosphokinase activity which transforms added ADP to ATP. In order to further characterize these reactions, the ADP-binding and nucleoside diphosphokinase activity of purified platelet membranes were solubilized by freeze-thaw injury followed by extraction with isotonic buffered saline. Up to 80% of membrane ADP-binding activity was solubilized along with 20% of the total membrane protein, a 4-fold purification. A Millipore filter binding assay was developed to detect the soluble binding protein using [3H]ADP as radioligand. Binding of [3H]ADP was rapid, reversible, saturable, and was destroyed by heat, trypsin digestion, and 1 mM N-ethylmaleimide. By Scatchard analysis, there was a single class of binding sites with a Kd of 3.8 x 10(-7) M. Unlabeled nucleotides competed with [3H]ADP with the following potency series: ATP = ADP greater than AMP greater than adenosine. The solubilized nucleoside diphosphokinase activity could be separated from ADP-binding activity by ultracentrifugation on 5 to 20% sucrose density gradients containing 0.6 M KCl suggesting that the activities reside on separate molecules. Hydrodynamic parameters were calculated for the binding protein by gel filtration and ultracentrifugation. The s20,w was 4.1, Stoke's radius 35 x 10(-8)cm, axial ratio (f/fo) 1.09, and the Mr = 61,000. The studies suggest that this platelet ADP-binding protein may act as the receptor for initiating ADP-induced aggregation and release.


Effect of trypsin on the cell surface proteins of hepatoma tissue culture cells. Characterization of a carbohydrate-rich glycopeptide released from a calcium binding membrane glycoprotein.

Concentrations of trypsin that bring about aggregation of hepatoma tissue culture (HTC) cells also release from the cell surface an Mr = 55,000 glycopeptide fragment. This glycopeptide fragment also accumulates in the medium, including serum-free medium, as a normal consequence of membrane protein turnover. The trypsin-released glycopeptide is labeled when cells are grown in the presence of fucose or leucine before treatment of the cells with the protease. Similarly, the glycopeptide fragment can be labeled by reacting cells in situ by lactoperoxidase-catalyzed radioiodination or by tritiated borohydride reduction of cells treated first with neuraminidase and galactose oxidase. The tryptic glycopeptide fragment was purified by concanavalin A-Sepharose chromatography, and hydroxyapatite chromatography in the presence of dodecyl sulfate. The amino acid and carbohydrate composition was determined, as was the sensitivity of the purified glycopeptide to a variety of endo- and exoglycosidases. The purified glycopeptide contains an average of 17 sialic acid residues and hence, shows charge heterogeneity after electrophoresis in isoelectric focusing gels. The charge heterogeneity can be eliminated completely by treatment with neuraminidase. The glycopeptide after this treatment is homogeneous. The trypsin-sensitive membrane glycoprotein which is the source of the Mr = 55,000 glycopeptide was identified by two-dimensional gel electrophoretic analysis of labeled cells, treated or not treated with trypsin. This glycoprotein, which has an apparent molecular weight of 85,000 and forms a homodimer in the presence of calcium ions, was purified and its identity as the parent of the Mr = 55,000 glycopeptide was confirmed by showing that the same Mr = 55,000 fragment was released by trypsin from the purified glycoprotein as was released from the intact cells.


Binding of androgen receptor to prostatic chromatin requires intact linker DNA.

Receptor-chromatin complexes were recovered from prostatic chromatin digested with micrococcal nuclease. The fragments of chromatin were separated on linear 7.6 to 76% (v/v) glycerol density gradients. With extensive digestion of DNA, receptor labeled with [1,2-3H]dihydrotestosterone was released from the chromatin. After 5% digestion of DNA to acid-soluble products, only a trace amount of labeled receptor was detected in the unbound form. In the latter instance, most of the labeled receptor was recovered from the gradients in association with five A260 peaks representing oligomeric and monomeric nucleosomes with a repeat length of 182 +/- 14 (mean +/- S.D.) base pairs. The concentration of receptors was highest in the A260 peaks, which contained large oligomers of nucleosomes, and lowest in fractions containing primarily monomer structures. Hence, the extent to which receptors remained bound to chromatin was dependent on the relative amount of intact, linker DNA present.