[The relationship between the state of the pulmonary circulation and indices of cerebral gas exchange in patients with acquired heart defects]. The author studied the state of hemeostasis in normals and in patients with acquired heart insufficiency. By means of bloody methods of triple probe samples of the blood inflow and outflow from the brain, and by biochemical methods they also studied 22 normals and 100 patients with acquired heart insufficiency for interrelations between the gas metabolism indices and acid-base equilibrium of the central pulmonary hemodynamics. As a results it was established that the brain blood flux in patients with acquired heart insufficiency in different degrees of pulmonary hypertension is adequate to the metabolic requirement of the brain in oxygen and corresponds to identical indices in normals. There was no relation between the state of pulmonary circulation and brain circulation.


Effect of laser irradiation and immunosuppressive treatment on survival of mouse skin allotransplants. The effect of laser beam on the survival of skin allotransplants has been studied. Irradiation of the donor skin and of the recipient graft bed improved the survival of the graft. The graft protecting effect of anti-thymocyte serum was considerably enhanced by laser irradiation.


Anesthesia for cesarean section III: effects of epidural analgesia on the respiratory adaptation of the newborn in elective cesarean section. A rise in fetal Paco2 was observed after elective cesarean section in patients anesthetized both with a barbiturate and with nitrous oxide/oxygen. Epidural analgesia seemed to be a good alternative in order to attain better blood gas values in the newborn infant. Fourteen healthy mothers and their infants were studied in connection with elective cesarean section. Epidural analgesia with plain bupivacaine 0.75% was used. Doses varied between 90 and 120 mg. The time between the epidural injection and delivery was around 50 min. In six cases the fetal heart rate was registered continuously. Most of the mothers were sedated with diazepam intravenously or fully anesthetized, after delivery. The mothers were interviewed later. The respiratory adaptation of the infants was studied by blood gas and acid-base measurements in repeated arterial samples during the first 3 hours of life. A comparison was made with a group previously studied, where general anesthesia with a barbiturate, nitrous oxide/oxygen was the method used. The present material showed no differences concerning Pao2 and Paco2 but clearly indicated a tendency towards an earlier normalization of the initial metabolic acidosis. Mothers showed a respiratory alkalosis which was overcompensated by the metabolic component. Maternal blood pressure falls were observed in four cases, and fetal effects could be detected. Although epidural analgesia has a more favorable effect upon the newborn's metabolic component, both the compared methods allow good respiratory adaptation provided they are used correctly. Mothers can be given the opportunity to choose between being conscious or asleep when their child is delivered.


The effect of halothane anaesthesia upon cerebral oxygen consumption in the rat. The influence of halothane (0.6 and 2%) upon cerebral (cortical) blood flow (CBF) and cerebral metabolic rate for oxygen (CMRo2) was studied in artificially ventilated rats, using a modified technique of Kety & Schmidt (1948). The values obtained in halothane anaesthesia were compared to those recorded in nitrous oxide anaesthesia, or to those measured in unanesthetized animals given an analgesic drug (fentanyl citrate). Although it could be confirmed that halothane induces vasodilatation in the brain, there were relatively small differences in CBF between the groups. The results demonstrate that, in the rat, halothane depresses CMRo2 in a dose-dependent way. With 0.6% halothane, CMRo2 was reduced by 20-30% and, with 2% halothane, CMRo2 was reduced by about 50%. Thus, in the rat the effect of 2% halothane upon metabolic rate is comparable to that observed in barbiturate anaesthesia.


Treatment of comatose patients by mechanical hyperventilation. In case of cranial trauma, early respiratory troubles either of central or peripheral origin often accelerate the deterioration of the neurological situation. The different values of PCO2, PO2, pH and alcaline reserve measured on samples of CSF in comatose patients prove the central acidosis related to metabolic and vascular disorders in the damaged areas. Our results confirm the correlation between the importance of this disturbances and the severity of the trauma. It is thus necessary to insure patients of satisfactory respiration conditions. The tracheobronchial cleansing is applicable to intubated or tracheotomized patients by an instillation of 5ml of simple or bicarbonated physiological serum 4 to 6 times a day, followed by repeated aspirations and associated to a preventive endotracheal instillation of 80 mg of Gentamycin 4 times a day. Moreover we use controlled respiration which does not modify the gazometric parameters in the CSF but which assures patients a normoxia and moderate hypocapnia with a decrease of intracranial hypertension. Treatment by controlled hyperventilation must be precocious, because the recuperation at the level of the damaged zones is very slow.


Resistant transitory epilepsy in children. Three case histories are presented which are characterized by progressive, severe symptoms of mental and epileptic nature with EEG abnormalities of increasing severity culminating in "hypsarhythmia". The seizures were resistant to any treatment over a period of several months. Apparently spontaneous cures followed, and the patients remained healthy during follow-up, which ranged from 10 to 12 years. The observations confirm Lennox's remark that a miraculous cure may occasionally be expected in even severe, probably symptomatic, epilepsy. Some exogenous (infectious?) cause is suspected.


A comparison of the cardiovascular effects of enflurane, halothane, methoxyflurane and fluroxene during open cardiac surgery. During open heart surgery hemodynamic changes due to 1.5% enflurane, 0.75% halothane, 0.18% methoxyflurane and 3.4% fluroxene have been compared. The following parameters have been measured: arterial pressure, heart rate, cardiac output, right and left atrial pressure, left ventricular pressure and dp/dt. The strongest effects were found with enflurane and halothane. Marked reduction in cardiac index, stroke index, left ventricular dp/dt as well as reduction of peripheral resistance caused severe systemic hypotension. Because of its slow uptake methoxyflurane was followed by small hemodynamic changes in this study. The rapid acting fluroxene had caused only minor reductions of cardiac output, stroke volume and dp/dt. There was no decrease in peripheral resistance.


Long-term hypotensive effect of atenolol (ICI 66.082), a new beta-adrenergic blocking agent. A report is given from an on-going multicenter trial in Sweden, in which 117 hypertensive patients have been treated with a new cardioselective beta-adrenergic blocking agent, atenolol (ICI 66.082, Tenormin) for an average of six months (range 2-21). Statistically significant reductions of BP were observed, recumbent by 29/19 mmHg (p less than 0.0001) and standing by 28/18 mmHg (p less than 0.0001). Few and comparatively mild side-effects were seen.


Bacteremia after tonsillectomy and adenectomy. 40% of negative hemocultures become positive immediately after tonsillectomies and/or adenectomies. This bacteremia is asymptomatic and remains no longer than one hour.


Hemodynamic long-term effects of timolol at rest and during exercise in essential hypertension. Sixteen men with previously untreated essential hypertension in WHO stage I have been studied as out-patients. Oxygen consumption, heart rate (HR), cardiac output (Q) (Cardiogreen) and intraarterial brachial pressure were recorded at rest in supine and sitting position and during steady state work at 300, 600 and 900 kpm/min. The subjects were treated with timolol as the sole drug for one year and the hemodynamic study was repeated. BP was reduced approximately 18% at rest and 14% during exercise, HR approximately 26% and the cardiac index 28% at rest supine and 32% at rest sitting. During exercise the reductions in Q were 25-30%. The calculated total peripheral resistance was significantly increased at rest as well as during exercise. The product of mean arterial pressure and HR was reduced about 40%. No severe side-effects were seen.


[Energy state of the cerebral cortex of the cat during hyperventilation (author's transl)]. Average Po2 and Pco2, local blood flow and pH values in the cerebral cortex of the cat were measured during passive hyperventilation (arterial Pco2 below 19 mm Hg). At defined intervals tissue samples were taken for metabolite analysis. The object of the study was to correlate the data obtained on the brain surface with metabolic responses. Immediately after the start of hyperventilation blood flow decreased, average cortical tissue pressures of O2 and CO2 fell, and there was a simultaneous rise in cortical pH. At a later stage in the experiment the local blood supply reverted to its resting level. Despite a fivefold rise in tissue lactate level during hyperventilation and a decrease in local O2 pressure on the brain surface to 5-10 mm Hg the degree of phosphorylation of energy rich phosphates was not less than under normal conditions of oxygenation. Our investigations showed no evidence of energy lack in cerebral cortex cells during hyperventilation. Cellular hypoxia and its characteristics are defined. The possible causes of raised tissue lactate levels during hyperventilation despite the lack of evidence of cellular hypoxia are discussed.


[Postoperative mental blocking in a continuous reaction task. With supplementary results showing the influence of age (author's transl)]. By means of a four colour device for measuring continuous reaction sequences the mental blockings of brain damaged patients in comparison with patients suffering from skin disease were determined. Differences due to age were also investigated. The analysis of frequency distribution of reaction times (dissection method according to Daeves and Beckel) yielded the following results: a) Brain damaged patients show a higher percentage of blockings (23%) than patients suffering from skin-disease (10%). "Normal" reaction times as well as "blockings" are not prolonged significantly. b) Older patients show prolonged normal reaction times and prolonged blockings without increase in the percentage of blockings. c) Patients with left hemisphere lesions show longer normal reaction times than those who undergo right hemisphere operations. The results are discussed with regard to their significance for theory and practice (road accidents).


Immunosuppression by fetal liver as a model for tolerance to self. The embryonic liver chimera system was used as a model to study the development of tolerance to self antigens. It was found that the permanent tolerant state which was induced in (C3H/eb x C57BL/6)F1 irradiated hosts following reconstitution with parental C57BL liver cells could be due to the development of suppressor cells within the liver cell inoculum, which specifically prevent reactivity of immunocompetent cells. General suppressor activity could be found in embryonic liver cells at early stages of gestation. However, the differentiation of such cells into specific suppressor of "self" antigens is dependent on the presence of the thymus.


Suppressor cells in the embryonic thymus mediate allograft tolerance. Thymus cells from non-immunized young chickens suppress the allograft rejection in lightly irradiated syngeneic or allogeneic recipients and mediate longlasting skingraft survival in a significant proportion of recipients across a strong histocompatibility difference. Suppressive activity of this kind is already found in the embryonic thymus and is therefore believed to mediate also self tolerance and neonatal allograft tolerance.


Internal mammary artery bypass graft in reoperative myocardioal revascularization. Thirty-two consecutive patients who earlier received indirect or direct myocardial revascularization underwent reoperation with one or more internal mammary artery grafts either alone or in combination with saphenous vein grafts. The main indication for reoperation was graft closure or progression of coronary atherosclerosis in nongrafted vessels, or both. Graft construction was performed under normothermic perfusion and anoxic arrest with interrupted suture technique. No intraoperative infarctions or hospital deaths occurred. All patients are alive after an average follow-up period of 20 months, and two thirds are asymptomatic. Arteriography after reoperation in nine patients revealed patency of eight of nine internal mammary artery and five of five secondary vein grafts. When angiographic and symptomatic indications for reoperation exist, the internal mammary artery bypass graft has become a valuable alternative, particularly for patients with small coronary vessels or previous vein graft failure.


Evaluation of a 5-mum stainless steel filter as an intravenous inline filter or prefilter. The suitability of a 5-mum stainless steel filter as an inline filter or a protective prefilter during simulated i.v. therapy was evaluated using flow rate measurements of two routinely used i.v. fluids and parenteral nutrition fluid. As an inline i.v. filter, the 5-mum stainless steel filter was capable of maintaining suitable flow rates. The addition of antibiotic additives decreased the flow rates slightly but not below the range required for i.v. therapy. Flow rate profiles, however, when compared to a 0.45-mum membrane filter suggest that antibiotic additives contain high numbers of particles in the less than 5-mum range. Consequently many of the particles, especially those in the less than 3-mum range will pass the 5-mum filter. As a protective prefilter, the 5-mum filter device in combination with a 0.45-mum membrane filter provided more uniform flow rates over longer periods of time when additives were employed. Using the aspiration device as a prefilter for adding antibiotics to the infusion fluid resulted in improved flow rates through a 0.45-mum membrane filter for lactated Ringer's containing cephalothin sodium, while for solutions containing ampicillin or oxytetracycline, prefilteration did not change the flow rate profiles.


Hyperactive children and the efficacy of psychoactive drugs as a treatment intervention. Characteristics of hyperactive children, including speculation in regard to etiology, are reviewed. Drug effects studies and drug treatment of hyperactive behavior are examined, and unresolved issues are discussed. Conclusions indicate that individual differences in hyperative children should form the basis for treatment planning, rather than simply treating groups of children under the rubric "hyperactivity."


Experiments on the role of virus infections in the pathogenesis of bronchial asthma. The role of innate or acquired insufficiency or ergotropic adaptation in the mechanism of genesis of bronchial asthma. The wide mosaic of congruent clinical and experimental observations led to the postulation that the cause of the pharmacological abnormality of the asthmatic patient, i.e. the immensely increased reactivity of the bronchial smooth muscles, is to be sought in an insufficiency of the beta-adrenergic receptor system. It is to be assumed that the so-called asthma diatheses is based inter alia on a genetically determined defect of the adenyl cyclase system. The role of previous infections of the respiratory tract in asthmagenesis should lie--following this working theory--not in a sensitization in the sense of an allergic reaction of the immediate type, but in the formation of a defective beta-adrenergic substance or in a blockade of the beta-receptor. A genetically determined innate defect of the beta-adrenergic receptors, or a defect acquired through infections of the respiratory tract, is hence likely to be the cause of the pathologically potentiated reactivity of the bronchia. It is likely that the infective stimuli--quite apart from this preparatory role--are later capable of triggering asthmatic paroxysms when the vegetative homeostasis is impaired. We know from the experiments of many authors that a blockade of the beta-receptors produced by chemical blocker substances, or by pertussis vaccine or various bacterial substances, results in a significant increase in bronchial reactivity towards histamine, serotonin, acetylcholine and other stimuli. We have shown in our experiments that heat-inactivated adeno viruses and influenza viruses also increase the anaphylactic shock reactivity and the histamine reactivity of the organism. On the basis of this working hypothesis, the pathomechanism of the asthmatic process is as follows in individual asthma forms: 1) In the 'purely" allergic asthma form, the antigen-antibody reaction that occurs after sensitization (i.e. formation of skin-sensitizing allergic antibodies of the class IgE) results in re-formation and release of slow-reacting-substances. spasm of the bronchial muscles, asthmatic paroxysm. The expulsion of catecholamines that follows the release of slow-reacting-substances makes a decisive contribution to the reestablishment of the impaired homeostatic balance. It is to be assumed that this form of asthma both symptomatically and causally--using specific desensitization--can be influenced more easily than other forms of asthma with a more complicated pathogenic background. 2) In the second allergically determined form of asthma, we are confronted by the genetically fixed or acquired insufficiency of the beta-receptors in addition to the immunological mechanism. As a result of the innate or acquired blockade of the beta-receptive substance, or the relative dominance of the alpha-receptors, the catecholamines (that physiologically serve to maintain homeostasis) contribute to a protraction, intensification and perpetuation of the bronchial obstruction. In this way the asthmatic circulus vitiosus is complete...


Ventilatory response to hypercapnia in the larger spotted dogfish Scyliorhinus stellaris. Dogfish were exposed to sudden changes of Pco2 in inspired seawater. During hypercapnia breathing frequency remained constant, but gill ventilation was transiently increased to about 140% of control levels in the 1st h. O2 uptake was significantly increased also, but returned to the initial level before nomalization of gill ventilation. In contrast to the transient rise in gill ventilation and O2 uptake, arterial Po2 was increased for the whole period of hypercapnia. Hypercapnia results in a marked fall in pHa which returned to the initial value in 4-5 h even though hypercapnia is maintained. This rise in pHa with little change in PaCO2 was associated with an increase in plasma bicarbonate concentration. The increase of plasma bicarbonate was in part due to compensatory bicarbonate uptake from the seawater across the gills and in part was effected by transfer between intracellular tissue compartments and extracellular spaces. The compensatory bicarbonate exchange mechanism in the gills seems to have a delay both after onset and termination of hypercapnia.


Does hyperinsulinemia in ob/ob mice cause an insulin-stimulated adipose tissue? After a 1-h preincubation to remove endogenous insulin, adipose tissue of obese mice (C57BL/L4 ob/ob) had a lower rate of glucose metabolism than tissue which was not preincubated. In contrast, preincubation did not change the metabolism of adipose tissue from lean mice (C57B1/6J +/+). The preincubation effect was abolished in obese mice which had had their serum insulin levels lowered toward normal by streptozotocin treatment. Injection of anti-insulin serum to obese mice caused adipose tissue removed 15 min after the injection to display a rate of glucose metabolsim lower than that of tissue removed before the injection. No such effect was seen in lean mice. These data are consistent with the hypothesis that hyperinsulinemia in the obese mice causes a chronic state of insulin stimulation of their adipose tissue, possibly contributing to their high rates of lipogenesis and their obesity. Several lipogenic enzymes were measured in adipose tissue of both lean and obese mice, and no single enzymatic abnormality was detected which might explain the hyperlipogenesis. Pyruvate dehydrogenase and acetyl-CoA carboxylase were both insulin-sensitive enzymes in lean and obese mice.


Importance of pulmonary ventilation in respiratory control in the bullfrog. Pulmonary and cutaneous O2 consumption (Vo2) and CO2 production (Vco2) were measured simultaneously in bullfrogs Rana catesbeiana at 20 degrees C. The lungs were responsible for 77.3-91.0% of the total Vo2 and 28.5-74.9% of the total VCO2. The distribution of the total exchange between the lungs and skin depended on metabolic rate; frogs with higher rates relied more heavily on the pulmonary mode for both Vo2 and Vco2. When prevented from ventilating their lungs in an O2-rich environment, bullfrogs developed severe respiratory acidosis, demonstrating the importance of lung exchange in normal acid-base balance. When frogs were totally submerged in an O2-saturated medium, skin Vco2 increased linearly to a steady-state value which approximated the preapneic total Vco2. In these same animals, arterial Pco2 increased proportionately to the increase in skin Vco2, indicating that skin diffusion capacity for CO2 was unaffected. We conclude that the control of breathing in the bullfrog in response to changes in metabolic rate relies predominantly on changes in lung ventilation while the skin plays a more passive role.


Electromyographic response of respiratory muscles during elastic loading. The response of respiratory motor neurons to graded elastic loading was assessed in anesthetized dogs by recording the electromyogram (EMG) from the diaphragm (ED) and the intercostal muscle (EIC). Elastic loads were applied for 1-20 breaths. The effects of changes in PCO2 on respiratory motor neuron output was assessed by applying loads during the course of CO2 rebreathing. On the first loaded breath, ED and EIC increased reflexly due chiefly to prolongation of inspiration. Vagotomy or vagal cooling to block the Hering-Breuer reflex eliminated the increase in ED and diminished the increase in EIC. During the second to fifth breath, the level of EMG activity was disproportionately high for the level of PCO2, suggesting an additional reflex component over and above the reflex activity present on the first loaded breath.


Plasma secretin and gastrin responses to a meat meal and duodenal acidification in dogs. In five conscious dogs with gastric fistula and two duodenal cannulas, plasma RIA secretin and gastrin levels were determined in response to 1) infusion of 0.1 N HCl in the proximal duodenal cannula, 2) ingestion of a meal, and 3) intraduodenal infusion of 0.1 N HCl following ingestion of a meal. Significant increases in plasma RIA secretin levels occurred during duodenal acidification. However, no significant change occurred in the secretin levels after ingestion of a meal, whereas significant increase in plasma gastrin level was observed. Postprandial duodenal pH remained above 4.5 for 3 h.


Intracellular pH of brain: alterations in acute respiratory acidosis and alkalosis. To evaluate the metabolic adaptations of the brain to acute respiratory acid-base disturbances, a method was developed to measure intracellular pH (pHi) in the brain of dogs under conditions in which arterial pH is rapidly altered. Brain pHi was determined by measuring the distribution of 14C-labeled dimethadione (DMO) in brain relative to cortical CSF. Brain extracellular space (ECS) was evaluated as the 35SO4 = space relative to cortical CSF, and arterial Po2 was maintained at 82-110 mmHg. In normal dogs, brain (cerebral cortex) pHi was 7.05, and after 1 h of hypercapnia (arterial pH = 7.07) it fell to 6.93. However, after 3 h with arterial Pco2 maintained at 85 mmHg brain pHi was normal (7.06), and during this time brain bicarbonate had risen from 11.3 to 24.4 meq/kg H2O. These changes were not prevented by intravenous doses of acetazolamide,


Myocardial CO2 buffering: role of transmembrane transport of H+ or HCO3-ions. Isolated rabbit hearts were perfused with rabbit red cells suspended in Ringer solution. A small volume of perfusate was recirculated for 10 min at Pco2 of 33.4 +/- 0.9 or 150.8 +/- 7.5 mmHg. Hypercapnia resulted in an increase in perfusate HCO3- concentration that was smaller than that observed when isolated perfusate was equilibrated in vitro with the same CO2 tensions (delta HCO-3e = 1.6 mM, P less than 0.01). This difference is consistent with a net movement of HCO3- into or H+ out of the mycardial cell, and cannot be accounted for by dilution of HCO3- in the myocardial interstitium. Recirculation of perfusate through the coronary circulation at normal Pco2 for two consecutive 10-min periods was not followed by changes in perfusate HCO3- concentration. A high degree of correlation (r = 0.81) was observed between intracellular HCO-3e concentration and the corresponding delta HCO-3e in individual experiments. The results suggest that transmembrane  exchange of H+ or HCO3- is a buffer mechanism for CO2 in the myocardial cell.


Cardiovascular effects of cerebroventricular ouabain perfusion in the adult dog. Cerebroventricular perfusion with artificial cerebrospinal fluid containing 10(-5) M ouabain was performed in adult dogs in order to describe the time course of the cardiovascular effect of intraventricular ouabain and to evaluate treatments to eliminate the cardiovascular effect. The central effect of ouabain caused a 56% increase in blood pressure above control values and a 35% increase in heart rate with various cardiac arrhythmias. Both alpha- and beta-adrenergic blocking drugs given intravenously.altered the pressure and rate effects ou ouabain, whereas vagotomy attenuated the effect.


Effect of beta-adrenoreceptor blockade on rat cardiac and skeletal muscle pH. The effect of catecholamines on the intracellular pH of rat cardiac and skeletal muscle during varying extracellular acid-base states was determined. Intracellualr pH (pHi) was calculated from the distribution of [14C]DMO. Acid-base disturbances were produced by placing the animals in an environmental chamber containing 10 or 20% CO2 or by administering HCL or NaHCO3. Two hours later the animals were anesthetized with sodium pentobarbital and blood and tissue samples obtained. In one series of animals, the effects of catecholamines were attenuated by administering the beta-adrenoreceptor antagonist MJ 1999 (Sotalol). In animals breathing 20% CO2, cardiac muscle pH was lower in beta-blocked than unblocked animals (6.69 vs. 6.78). During metabolic acidosis, cardiac muscle pH of beta-blocked animals was lower than that of unblocked animals (6.75 vs. 6.84). The same relationship was observed for skeletal muscle during metabolic acidosis-beta blockade pHi, 6.66; unblocked pHi, 6.77. The pHi of beta-blocked versus unblocked animals was not significantly different under normal acid-base conditions or metabolic alkalosis for cardiac or skeletal muscle. The effective buffer value of both tissue over the normal acidotic range was decreased by the beta-blocking agent. These results indicate that catecholamine release accompanying acidosis attenuates the change in pHI and increases the effective buffer value of cardiac and skeletal muscle.


Biliary secretion in elasmobranchs. I. Bile collection and composition. Bile composition and secretion were studied in tow elasmobranch species, Squalus acanthias (spiny dogfish shark) and Raja erinacea (small skate), after ligation of the common duct and insertion of cannulas into the gallbladder lumen. Fish were then allowed to swim freely in large pools and bile was collected in balloons attached to the externalized cannulas. Both species secreted bile for periods of 4-7 days at a maximum rate of 1.77 +/- .89 ml/kg per 24 h in Squalus acanthias and 2.66 +/- .89 ml/kg per 24 h in Raja erinacea. Comparison of the composition of cannula and gallbladder bile indicated that hepatic bile could be collected by this technique in both species without effective contact with gallbladder epithelium. The low concentrations of HCO3- and CL- and the gigh bile salt levels in dogfish gallbladder bile indicate that mechanisms for concentration and acidification of bile in the gallbladder developed early in vertebrate evolution. These results indicate that gallbladder cannulation in free-swimming elasmobranchs is a useful technique for the study of bile secretory and excretory function in marine species.


Film screening by physician assistants in diagnostic radiology. Data are presented which indicate that specially selected ART students can be trained to evaluate radiographs as to the presence or absence of significant pathology with an accuracy equivalent to that of experienced staff radiologists. The methods employed in this training program-namely, stressing perceptual recognition of normal and abnormal findings-may be beneficial in the early months of training of diagnostic radiology residents to hasten achievement of an acceptable perceptual performance. Test films, used in this study as an evaluation technique, may also be useful to assess residents' progress.


Use of physician's assistants in a general surgical practice. A study has been made of three PAS employed in a rural, small town, general surgical practice during a four year period. Their work was equally divided among three areas: minor trauma, obtaining and recording data for history and physical examinations, and assisting at routine surgical procedures and endoscopies. This resulted in a significant saving of time for each surgeon who was thus freed for more time in the care of the more critically ill or injured patients.


[The influence of PaCO2 on oxygen consumption during extracorporeal circulation in hypothermia (author's transl)]. Hypocapnia during extracorporeal circulation in hypothermia increases oxygen consumption. Po2 in mixed venous blood decreases. This probably reflects a decrease in tissue oxygen tension. Hyperventilation will therefore increase the risk of hypoxia in critically perfused tissues. Therefore we recommend to keep PaCO2 (T) constant at 40 mm Hg during hypothermia.


[Clinical studies on the use of ketalar in obstetric anesthesia (author's transl)]. 26 healthy pregnant women at term were divided into 2 groups and anaesthetized with either 1 mg/kh or 2 mg/kg ketamine - N20/02 - for primary caesarean section. Maternal venous ketamine plasma levels, neonatal ketamine plasma levels (umbilical artery and umbilical vein) and blood gases were measured. Maternal venous ketamine plasma levels in group I exceeded those in group II by 2.8 (30-60 sec after injection) and 2.0 (at the time of delivery) respectively. Independent of the dose used, the plasma levels of ketamine in the umbilical artery or the umbilical vein were found to be identical in the two groups of newborn infants. Neonatal blood gases and acid base parameters did not significantly differ between the two groups, except for the oxygen tension in group II which slightly exceeded the PO2 values in group I 15, 60 and 120 min after delivery. pH and standard bicarbonate values were found to be higher (1 and 5 min) in both groups, compared to pH and standard bicarbonate levels in a group of newborns delivered spontaneously.--The results of this study show: 1. Independent of a low or high ketamine dosage, neonatal blood concentration of the drug remain low, probably due to a placental barrier effect.--2. The post partum recovery of the newborns was neither influenced by the ketamine anesthesia itself nor by different drug doses.


Effect of abdominal compression on minute ventilation of patients with chronic obstructive lung disease and bronchial asthma. The effects of manual compression of the upper lateral abdomen (C) of 73 cases of obstructive airway disease were studied in respect to the change in minute ventilation (V) 10 minutes after the procedure was performed. A mean decrease of 13% in V was found in the majority of 15 cases of bronchial asthma or chronic bronchitis and 43 patients with chronic obstructive lung disease. The use of manual compression as a therapeutic measure depends largely on the abrupt reduction of lung volume in cases of obstructive airway disease in which over-inflation of the lung is present as an acute or chronic disorder. The degree and duration of clinical benefit, i.e., relief of dyspnea, depends on such factors as the volume of air trapped in the lungs, broncho-constriction through pathologic change or bronchospasm and impairment of elastic recoil of the pulmonary parenchyma. The lowered V recorded 10 minutes after C is performed suggests that decrease in dyspnea is associated with enhanced efficiency of ventilation in some subjects with obstructive airway disease, including those with bronchial asthma as well as COLD. A brief description is presented of 10 patients with COLD in whom manual compression and other features of a rehabilitation program were employed. Mean length of life of five living subjects is 12.6 years after beginning of therapy. The mean length of life of five who died was 10.2 years after beginning of treatment.


[Accidental peroperative hypothermia during rapid transfusion]. Six cases of grave hypothermias are reported, having arisen during surgical interventions which necessitated a rapid and abundant transfusion of badly warmed blood. The role of favouring factors, surrounding cold due to the air-conditioning, anaesthesia, extent of the area of operation, seems important. The symptomatology permits the individualization of a hypothermic syndrom neighbouring the picture described in toxic accidental hypothermias. Accidents during the warming process associate collapse and disturbances in coagulation. It is therefore necessary to consider certain signs of alarm as important and generalize the conditions for prevention of thermolysis in the operating theatre.


[Information science in intensive care medicine. Examples of the European approach]. The modern treatment of patients in a precarious state requires the use of technical appliances which become more complex and more numerous day by day. The data generated by these appliances, together with the conventional measurements of physiological parameters carried out by the nursing ataff during care, with the results of laboratory examinations and with medical physical examinations, represent a raw material of information which tends to increase each year. The repetitive and hand-written consigning of these data, likewise their traditional laborious consultation, no longer guarantees today the taking of rapid decisions concerning the patients. Now, this rapid decision-making is at the centre of the functioning of present-day intensive medicine, of the surveillance and optimal treatment of the patients, of the functional organization of the intensive care unit, and of the "economic" efficiency of the exploitation. It seems locigal, therefore, to wish to avail of the resources provided by information science to resolve these problems. The United States, Japan and certain European countries have done so. Our purpose is to give some examples of the European approach.


[Iskedyl and general anesthesia. Measurement of the systolic ejection volume and peripheral resistance using the arterial pressure curves]. ISKEDYL (PF 50), which is sold as a vasoregulator of cerebral irrigation, consists of a mixture of dihydroergocristine and raubasine. This work aims at determining possible interactions between this product and certain drugs used in anesthesia from the cardiovascular point of view. ISKEDYL does not seem to be a contraindication to anesthesia of the "neuroleptic" type, when injected in pre-, per- or post-operative period. A slight temporary and spontaneously reversible fall in arterial blood pressure, affecting both maximum and minimum pressure, together with a decrease in stroke volume and peripheral resistances, estimated by the study of the arterial blood pressure curve, seem to indicate that this product has peripheral vaso-dilatory properties.


[Continuous peridural anesthesia in traumatological and reparative surgery of the lower extremities]. The authors give an account of their experience of peridural anaesthesia in 250 cases at the Centre for traumatological and reparative surgery. They demonstrate the advantages derived from this type of anaesthesia: in elderly subjects manifesting an organic defect, and in emergency cases with "full stomach".


[Disorders of body temperature regulation. 1. Physiopathological study]. In this work, disorders of thermo-regulation are first studied in a synthetic manner from the angle of cellular and haemodynamic disturbances. Then the particular physiopathological aspects concerning hypothermia and hyperthermia are looked at in succession. In his next work, the author will view the clinical and therapeutic study of these phenomena. Extensive bibliography.


[Physico-chemical characteristics, absorption and elimination kinetics and technics of use of enflurane]. Physical-chemical properties, uptake und elemination of enflurane are described with particular references to halothane. The ratio potency/vapor pressure, the blood and tissues solubility of this agent provide a good flexibility of uptake and elemination in response to changes in alveolar ventilation. When used with nitrous oxide, the enflurane may provide good clinical results at inspired concentration 1-2 p. 100.


[Impact of alfatesine anesthesia on cerebrospinal fluid pressure in man]. The variations in pressure of the cerebrospinal fluid was studied in 20 patients before and after induction of anesthesia with alfatesine (0.1 ml/Kg). 14 patients received no other complementary drug (group 1); in the six other cases, 1 g. of acetylsalicylic acid was administered as an analgesic complement (group II). All of the patients spontnaeously ventilated an O2 - N2O 50 p. 100 mixture. The C.S.F. pressure fell by 39 p. 100 on the average (p. less than 0.001) in group I and did not vary in group II. This fall is essentially related to cerebral vasoconstriction, therefore to the fall in cerebral blood flow caused by Alfatesine. In group II hypercapnia was noted in all of the patients; it abolishes the cerebral vasoconstriction due to Alfatesine; the cerebral blood flow did not fall neither did the C.S.F. pressure.


[Transformation of sorbitol to mannitol when used in parenteral administration]. This was shown by the unexpected discovery of the urinary excretion of mannitol when urinary sorbitol levels were being measured in connection with the surveillance of patients suffering from malnutrition and head injury and receiving total parenteral nutrition: glucose, sorbitol, amino-acids, soya oil. The presence of mannitol in the urine would appear to result from a secondary catabolic through physiological pathway for the fructose produced by the metabolism of sortibol not metabolised during glycolysis.


[Clinical evaluation of the new anesthetic "Ethrane"]. Seventy-four patients aged 14 months to 71 years, classified as ASA I and II were anesthetised with Ethrane for surgical interventions of mean duration 117 minutes. With the exception of 5 patients who were directly anesthetised with Ethrane, the others received Ethrane after induction with Penthiobarbitone. Maintenance of anesthesia was ensured with 1 to 4p. 100 concentrations of Ethrane and 33p. 100 oxygen and 66p. 100 nitrous oxide. Tracheal intubation was facilitated by injection of 1 mg/kg of succinylcholine. Induction with enflurane is rapid with no phenomena of excitation or irritation of the ear passages. The cardiovascular apparatus is stable with no arrythmia but an increase in heart rate of 11 to 50p. 100 is noted and in 41p. 100 of the cases hypotension of 35p. 100 of the intitial value. During spontaneous ventilation, a type of rapid and superficial respiration is observed with a flow volume of 5.3 ml/kg for an average frequency of 25/min. The arterial blood gases show slight hypercapnia. Myorelaxation is significant and better than that obtained with halothane. Coming round poses few problems apart from agitation in adolescents. Response to simple orders appears at 13 minutes. Trembling and rigidity occur in 41p. 100 of the cases for 5 to 30 minutes. From the hepatic point of view, no lastin enzyme changes were noted and no renal toxicity was demonstrated. Ethrane appears to be a good anesthetic agent but the few advantages mentioned means that it does not fulfil ideal conditions.


[A case of acute tobacco poisoning by enema]. The authors report the case of a 5 month old child intoxicated with nicotine through an enema. A mother who wished to treat her infant for constipation had administered to him as an enema a decoction of green tobacco leaves; 15 to 20 minutes after the enema, the child showed diarrhoea and respiratory disturbances. The authors treated the respiratory disturbances by assisted respiration and bradycardia and the diarrhoea with atropine. They also caused a force diuresis using a 10 p. 100 glucose solution and Furosemide.


[Hemodynamic effect of enflurane in man]. The following hemodynamic parameters: cardiac frequency, peripheral arterial pressure, pulmonary pressure and cardiac output were measured by direct catheterisation, as the total peripheral vascular resistance and the systolic ejection volume were calculated from the registered results. The cardiac frequency and the pulmonary arterial pressure were practically not modified in our patients, though we have observed a statistically significant decrease of systolic (-30p. 100) and diastolic (-27p. 100) arterial pressure. The total peripheral vascular resistance shows a marked diminution (-20p. 100) after giving Ethrane? for ten minutes. If it is possible that one part, surely important, of the cardiac output, is preserved under Ethrane anesthesia by a significant decrease of the total peripheral vascular resistance, a myocardial depression might be questionned, the decrease of cardiac output at 30 minutes being more important than the decrease of the total peripheral vascular resistance.


[Postoperative circulatory arrest (apropos of 37 cases)]. Within five years, the authors received 37 patients who had presented a post-operative cardiac arrest. Most of them came from the Bordeaux district (from the hospital as well as the public sector) although they did not represent the totality of such accidents. The study bears first of all upon the data concerning the patients, then upon the surgical intervention, the mode of anaesthesia and ventilation, the posture and finally the treatment. In 10 cases. it was noted, as early as the admission, that the blood volume feel short of the theoretical total blood volume by one liter or more. The authors take into account how traces of previous affections, the localization in O.R.L., and, in ophthalmology, an important number of mistakes in anaesthesia, favour the progress of the genesis of the accident.


[New ventilation technic in anesthesia for direct suspension laryngoscopy]. The authors bring not only the results of their experience but a solution of the respiratory problems during anaesthesia in suspension laryngoscopy, thanks to the devising of a new intubation probe and a pre-set automatic insufflator with varying volum flow.


[The second electromyographic study of surgical curarization with AH.8165, a new pachycurare derivative of azobis-arylimidazo(1-2a) pyridinium]. This second electromyographic study of AH-8165, a product derived from azobis-arymilidazo-(1-2a) pyridinium, carried out with a special apparatus which both stimulates and records, specifies the characteristics of the neuro-muscular block induced by this new non-depolarizing type curarizing substance while taking into account the usual factors of the curarimimetic variability of action. After an initial dose of 1 mg per kg of bodyweight, we notice particularly: -the lapse of time required for a complete block (90 s), its intensity and duration (normally total during at least one hour) -the morphology of the electromyogram during the curarization and the decurarization either spontaneous or induced by Neostigmin with in particular -the muscular fatigability after repeated stimulation following curarization and chiefly during decurarization, chronological data und electro-myographic aspects which are found as well after a reinjection of AH-8165 equivalent to the half of the previous one.


[Contribution to the study of Trive 1000]. A glucid-lipid and protein compound nutriment: Trive 1000 was administered to 53 patients during the first three postoperative days in varying doses: -1.000 ml per day to a first group -1.500 ml per day to a second group. The local tolerance was quite good. However, a few accidents of a general character were noted: they were due to a great extent to a non-observance of the rules of preservation. A study of the glucides, in Sorbitol form, showed a good assimilation of the substratum. Glycemia remaining within normal, no glycosuria or ketonuria could be observed. The assimilation of the proteins seemed on the whole satisfactory. No important rise in the blood urea was noted. Nitrogen balances showed that, thanks to this nutriment, almost half of them were positive during the post-aggressive period. The more important the dose of Trive 1000, the more positive the nitrogen balances were. The administration of lipids did not reveal any significant change in the total lipids, triglycerides or cholesterol. A study of the graphic record of lipid levels showed chylomicrons twice. On the opposite, lipoproteins were hardly modified apart from a significant in pre-beta lipoproteins. The quantitative analysis of amino-acids proved interesting. A comparative study of amino-acids both in blood and urine before and after the administration of Trive 1000 showed very little variations in connection with essential amino-acids. If the proportions in the blood of non-essential amino-acids were comparatively but little modified before and after the treatment, on the oppostie, the urinary clearance rate of the same amino-acids after treatment was far more variable and often quite important as far as certain amino-acids were concerned.


[Comparative study of nitrogen savings during enteral and parenteral feeding (Trive 1000)]. Through a study of Nitrogen balances, the authors compare the effectiveness of a proteinic intake carried out per os or by intravenous route. The study involved 22 different carcinoma carriers and lasted for 9 days during which the Nitrogen intake was effected either by the digestive tract or by intravenous route supplemented by a sufficient caloric ration. In the latter case, the aminoacid intake was supplied by Trive 1000 combined with glucides and lipids which were then quantitatively deducted from the enteral intake. A study of the results showed that the assimilation of the Nitrogen injected intravenously was very similar to the one obtained by enteral route.


[The utilization of EB 51 in parenteral feeding. Clinical, biological and anatomopathological control. Statistical study on 30 patients]. Two groups of patients (comas of central origin and serious digestive undernourished) were submitted to an exclusive or a supplementary parenteral feeding for a short or a long period of time totaling over 500 days. The catheter was placed (in 80 p. cent of the cases) in a deep vein and was tunnellized. EB 51 (Trivemil) used in those 30 patients gives amino-acids, lipids and glucides. The caloric intake was completed by 30 p. cent glucose solutes. If the average caloric intake was 3.000 calories per day, it was often above that figure since the feeding was combined with a nutri-pump enteral feeding for 60 p. cent of the days. The clinical tolerance was quite good. On a biological level, in addition to classic investigations, 40 analytic graphic records of lipid levels were made, as well as a quantitative analysis of serous triglycerides and lipurias, a study of sorbitol and 500 nitrogen balances combined with 150 chromatographies on the acidaminuria column. An hepatic biological control was also made (amnoniemia, transaminases) as well as a quantitative analysis of lactates, pyruvates and minerals: iron-phosphorus-magnesium. Some of the results were checked by statistical studies. No serious anomaly was detected. Lung and hepatic biopsies carried out upon 9 patients did not apparently reveal any lipidic overloading. The clinical and the biological investigation show that this complete nutriment is perfectly tolerated and assimilated.


[Effect of increasing doses of dopamine on the left ventricular function in the dog]. A study on eleven dogs of the effects of increasing doses of dopamine on the left ventricular function. A description of the method which consists of a venous shunt connected to an extra-corporeal circuit and which allows modification simply and rapidly of the state of vascular refilling of the animal. The results differ according to the pressure-level of the refilling of the left ventricule. At low pressure, the dopamine increases the arterial pressure, the cardiac output and the systolic activity of the left ventricle for a reduced tachycardic effect. At higher pressure, the average aortic pressure is only slightly increased and the systolic activity is elevated without increase in cardiac output. These facts indicate dopamine in states of shock with a low pressure of refilling.


[The effect of intravenous dopamine on the hemodynamics of the heart]. The pharmacological study of dopamine was conducted on 14 patients: eleven normal patients and three with incipient myocardiopathies. The dosages used were 3, 6 and 12 mug/kg/min. The "pump" function, the peripheral resistances, the contractility and the ventricular compliance were studied. Dopamine is a positive inotropic agent without chronotropic action at doses of 6 and 12 mug/kg/min. It acts by increasing the contractility and the venous return and by decreasing the peripheral resistances; this effect disappears with strong dosages.


[Hemodynamic study of dopamine used in chronic heart failures and in cardiogenic shock as a complication of acute myocardial infarct]. The object of this study is to examine the properties of dopamine at the hemodynamic and renal level in 16 patients with decompensated chronic cardiopathies or very serious cardiogenic shock due to myocardial infarction. The results show an increase in the cardiac index in 75 p. 100 of the cases with a favourable diminution of the arterio-venous difference in O2 and of the pulmonary arterial resistances. The most noticeable and the most constant effect is the recovery and increase of the diuresis. Positive chronotropic and bathmotropic effects were observed which necessitated the limitation of prescription of dopamine in those subjects showing signs of disturbances in ventricular excitability.


[Studies of the variations in functional renal values induced by intravenous dopamine]. We studied, on six patients, the variations in functional renal value and urinary excretion of electrolytes during the intravenous administration of dopamine and in the hours immediately following. Two groups of patients are distinguished. In groupe I, all of whom had normal functional renal values, there was no modification of these, while we observed increases in output, in excretion of electrolytes and in the clearance of the uric acid. In group II, composed of patients with renal insufficiencies, the modifications are less definite but the diuretic and saluretic effect is present. The salidiuretic effects of the dopamine would seem to be dissociated from the cardiovascular effects.


[The use of dopamine during postoperative cardiogenic shock in children. Preliminary results]. Dopamine was used in a dose of 5 mug/kg/min in ten infants with congenital cardiopathy and presenting in the immediate postoperative period a syndrome of low cardiac output. The output was not measured but, based on the evolution of the clinical signs, six favourable results, with correction of the syndrome, can be reported.


[Use of dopamine in postoperative reanimation after heart surgery. Preliminary results]. The dopamine was used on ten patients having undergone one or several valvular replacements under extra-corporeal circulation. The essential indication was the appearance postoperatively of more or less serious circulatory failure. The dopamine was administered by drip in doses of 2.5, 5 or 10 mug/kg/min. The effects on the frequency of the cardiac rhythm were moderate. On two cases, ventricular hyperexcitability induced by isoprenaline disappeared under dopamine. The chief effects were an increase in cardiac output, in the form of an increase in the volume of systolic ejection and lowering of the peripheral resistances. A steady increase in urinary volume preceded the amelioration of clinical signs of circulatory failure.


[Use of dopamine in acute cardiovascular distress. Its place in comparison to isoproterenol]. Nineteen patients presenting cardiovascular distress were treated with dopamine. For seventeen of these, the accident occurred immediately after cardiac surgery. Dosages varied from 1 to 15 mug/kg/min and the duration of treatment from 10 minutes to three days. The efficacy of the treatment was judged according to the clinical and hemodynamic improvement of the cardio-circulatory function and the increase in urinary output. There were 15 favourable results. The positive effects of the dopamine seem to be limited in certain patients by the appearance of a cyanosis testifying to a rise in vascular resistances which increases the left auricular pressure and limits the inotropic effect. In these cases, isoproterenol or a combination of both isoproterenol and dopamine gives better results.


[Dopamine and shock. Preliminary clinical study of 7 cases]. Seven patients presenting a state of shock were treated with dopamine. The authors remark an undeniable effect on arterial pressure which rises again and on the diuresis.


[Use of dopamine in the treatment of cardiogenic shock. Preliminary results]. A study of seven patients, each of whom was treated with dopamine within three hours after suffering a myocardial infarction. For four of these, a comparative study was made with isoproterenol, glucagon and ouabaine. The average age of the subjects was 72 years, and all presented considerable myocardial lesions before the treatment was begun. Despite improvement, particularly in diuresis and cardiac output, none of the patients survived. The authors explain these results by the fact that, like all powerful inotropic agents, dopamine produces an increase in oxygen consumption of the myocardium for the ischemic cells situated in the zone contiguous to the infarct.


[Comparison of the endocrine response under 2 kinds of anesthesia: neuroleptanalgesia of the chlorprothixene-dextromoramide type and venous anesthesia of the type alfadione-fentanyl]. In 14 patients anaesthetized before undergoing an orthopedic surgical intervention, the variations induced by anaesthesia in the 17 hydroxycorticosterone rate, catecholamine, somatotropic hormone (STH), insulin, glycemia, free fatty acids and thyrotropin (TSH), all these variations were studied before the surgery. The patients were divided into 2 groups of 7, the first one being anaesthestized by chlorprothixene dextromoramide Neurolept-Analgesia and the second one by Alfadione Fentanyl venous anaesthesia.


[The place of enflurane in neuro-anesthesia]. In neuro-anaesthesia, anaesthetic agents must be in accordance with certain criteria necessary to the preservation of the integrity of the brain. Therefore it was in such a perspective that we re-appraised the effects of enflurane upon cortical irritability, metabolism, cerebral blood flow and intracranial pressure. We reached the following conclusions: in normocapnia, and with optimal clinical concentrations, this drug remains the best anaesthetic agent since it has no harmful effect either upon metabolism or upon cerebral blood flow. However, in hypertensive cerebral lesions, caution is called for an it seems advisable to combine Enflurane with mild doses of fentanyl. Finally, anaesthesia with enflurane is followed by a rapid and smooth return to consciousness, a valuable factor in neurosurgery where post-operative neurological watching matters very much.


Immunology and microbiology in acute otitis media. Various immunological parameters were measured in serum, middle ear fluid (MEF), and lymphocytes from peripheral blood and MEF of infants with acute otitis media due to S. pneumoniae or H. influenzae. Approximately half of 131 patients had IgE specific antibody to the infecting bacterium as determined by the indirect fluorescent antibody (IFA) technique. Seventy-one percent of these IgE positive patients had IgE specific antibody in the MEF. Total IgE concentration was found to be from an average of 1.5 to 3.0 times higher in the MEF when compared to the simultaneously drawn serum. In addition, antibody to pneumococcal capsular polysaccharides and to pneumococcal C-carbohydrate was demonstrated in the MEF by radioimmunoassay. When MEF specific antibody was compared to serum antibody it appeared that antibody to C-carbohydrate was more concentrated in the MEF. That this antibody was of the IgE class was suggested by IFA but not conclusively proven. Evidence exists that conditions for enhanced IgE synthesis is concomitantly associated with a decrease in T-cell activity. T-cell function in MEF derived lymphocytes as determined by rosette formation and by phytohemagglutinin (PHA) stimulation was approximately one-tenth that of the peripheral blood lymphocytes. However, that T-cells may participate in the immune response to polysaccharides was suggested by the observation that polysaccharide stimulated peripheral blood lymphocytes from infants immunized with octavalent pneumococcal capsular vaccine underwent protein synthesis two to three times that of the PHA stimulated cells. The clinical significance of this finding as well as the nature of the cell responsible for the increased protein synthesis remains to be established. It is hypothesized that acute otitis media results from local synthesis of bacteria specific IgE antibody which is enhanced by a paucity of local T-cell activity.


Otitis media in the newborn infant.

Otitis media in the newborn infant. Otitis media occurs frequently in newborn infants and often is associated with systemic infection. Patients studied at the Boston City Hospital have provided preliminary data on the otoscopic findings, tympanometric patterns and etiologic agents characteristic of otitis in this age group.


Microorganisms in chronic otitis media with effusion. A total of 175 effusions obtained from 172 patients suffering from chronic otitis media with effusions was examined for bacterial smear and culture. Eighty percent showed positive bacterial smear, but only 49% yielded positive bacterial culture. The mucoid effusions had positive cultures in only 37%, whereas the bacterial culture rate was higher in serous (59%) and leukocytic (64%) types. The isolation of common pathogens accounted for about 50% of the isolates, and nonpathogens accounted for the remaining 50%. The high incidence of microorganisms in the middle ear effusions in the present series raises the possibility of bacterial contribution in many cases of OME.


Gastro-oesophageal reflux and hiatal hernia. A re-evaluation of current data and dogma. Various methods of investigating and treating patients with gastro-oesophageal disorders are described and the rationale of current concepts is outlined. Emphasis is placed throughout on gastro-oesophageal reflux and its sequelae rather than on sliding hiatal hernias. Symptoms of gastro-oesophageal dysfunction can be misleading, and careful studies are essential in assessing its importance and the results of various modes of therapy.


A new look at the measurement and interpretation of enzyme assays. Some methodological problems in clinical enzymology, including instability of enzymes in the incubation mixture and requirements for optimal reaction conditions, are highlighted. The importance of a knowledge of fundamental enzyme biochemistry and physiology as the basis for their diagnostic application is stressed, and the different behaviour of some hepatic enzymes--namely, GOT, GPT, gamma-GT, and OCT, in various pathological conditions is traced back to their characteristic biochemical and physiological properties. In the field of urinary enzymes a knowledge of the ideal requirements for the enzyme investigation of the various renal functions and of the properties of potentially valuable enzymes permits a critical selection of the really useful ones.


Allergic factors in management of middle ear effusions. There is no single specific therapy for the treatment of middle ear effusions and, therefore, a holistic approach to management is necessary. Current management involves multiple therapeutic modalities of which we believe tube and allergy are most efficient.


[Properties of penicillin amidase covalently bound to cellulose matrices]. Properties of penicillinamidase (PA) covalently bound with the cellulose matrix were studied. The efficiency of the binding depended on the bind type and purity of the native enzyme taken for binding. Stability of the immobilized PA (IPA) was studied at wide pH ranges. The effect of the ion strength, substrate concentration and purity of the native PA on stability of IPA was also investigated. The maximum stability of the enzyme was observed at pH 6.5-7.0 Stability of IPA depended on the purity of the native enzyme. When PA of the diazotized ether of cellulose containing amino groups was used, the enzyme was destabilized. IPA prepared on chlortriazinylcellulose was more stable than the respective native PA almost by I order.


[Determination of the biological activity of heliomycin by a method of diffusion in agar]. A biological method for determination of geliomycin activity using agar diffusion is described. A nutrient medium containing Hottinger broth up to 35 mg per cent of amine nitrogen, 1.5 percent of agar-agar, tap water, pH 7.8 to 8.0 was used. Bacillus subtilis ATCC 6633 served as the test-culture. Geliomycin was dissolved in 0.1 N sodium hydroxide solution.


[Study of the effect of the oxytetracycline crystallization conditions on the process indices]. Such factors as the rate of the changes in pH, temperature, mixer speed and the nature of the anions present in the solution has a significant effect on the indices of oxytetracycline dihydrate crystallization, i. e. residual content of the antibiotic in the mother solution and the specific surface of the crystalls. In this connection the effect of the above factors on the main indices of the process were studied. On the basis of the experimental data dependences were found which provided determination of the crystallization conditions securing the process indices.


[Analysis of the data in the literature on the sensitivity of cholera vibrios to tetracycline]. Variation diagrams were proposed to be used for the analysis of the published data on the study of Vibrio cholerae sensitivity to tetracycline. The diagrams provided systematization of dissimilar data of the experimental studies and a unique system of estimation was created. The analysis of the systematized data provided dividing of the Vibrio cholerae strains tested into 4 groups differing in the levels of their sensitivity to tetracycline. The groups were independent of the Vibrio cholerae biotypes and the place and period of the strain isolation.


[Toxicological characteristics of ampicillin]. Toxocity of ampicillin trihydrate was studied in acute and chronic experiments. It was shown that the antibiotic had low acute toxicity, did not cumulate and had no skin-irritating effect. On its inhalation in concentrations of 5 mg/m3 for 4 months, ampicillin induced allergization of albino rats, decreased their immunity. The general toxic effect of the drug was slightly pronounced. Ampicillin in a concentration of 0.1 mg/m3 induced tension of the immunological reactivity of the organism. The maximum permissible concentration (MPC) of ampicillin in the working premises equal to 0.1 mg/m3 is recommended. Mark "Allergen" is necessary.


Susceptibility of pneumococci and Haemophilus influenzae to antibacterial agents. Strains of Diplococcus pneumoniae and Haemophilus influenzae were tested for susceptibility to numerous antibiotics by a twofold agar dilution method using an inocula replicator. Undiluted, fully grown broth cultures were used as inocula for both species, and cultures of pneumococci diluted 1:1,000 were also tested. The antibiotics included most of those in common use in the United States as well as some chemical modifications recently approved and others that are under investigation. The most striking aspect of the results was the marked susceptibility of the pneumococci to all the antibiotics tested except the polymyxins and most of the aminoglycoside antibiotics, although some new aminoglycosides were active in quite low concentrations. Some of the strains of pneumococci were of decreased susceptibility to penicillin G (minimal inhibitory concentrations, 0.2 to 0.4 mug/ml), but none were tetracycline resistant, although such strains had been reported previously from this laboratory. The strains of H. influenzae, which were all serologically nontypable, exhibited different patterns of susceptibility to the groups of antibiotics and to the individual chemically related ones. None of these strains (isolated early in 1972) were ampicillin resistant. The most active agents against H. influenzae were: carbenicillin and ampicillin, analogues related to each of them, rifampin, chloramphenicol, and the polymyxins. However, the tetracycline analogues other than tetracycline, some aminoglycosides, notably tobramycin, kanamycin, gentamicin, and verdamicin, erythromycin, and some new lincomycin analogues were also active in low concentrations. Trimethoprim alone was highly active, and in combination with sulfamethoxazole it was even more active and synergistic against strains of both D. pneumoniae and H. influenzae.


Comparative incidence of phlebitis due to buffered cephalothin, cephapirin, and cefamandole. Buffered cephalothin, cefamandole, and cephapirin were compared with respect to their tendency to produce phlebitis. Two grams of each agent was administered every 6 h for 4 days to 12 healthy volunteers in a double-blind crossover fashion. Approximately 50% of intravenous sites developed mild (grade 1) phlebitis and 25% developed moderate (grade 2) phlebitis. The frequency of grade 1 inflammation did not differ significantly among the three cephalosporins. The proportion of individuals eventually exhibiting grade 2 phelebitis was highest with cefamandole, lowest with cephalothin (P = 0.07), and intermediate with cephapirin; however, cephapirin required a substantially greater number of doses to produce grade 2 phelebitis than did the other two drugs. These findings, together with the results of other reports, suggest that interpretation of the phlebitogenic potential of these antibiotics must be made with caution.


Citric acid metabolism in hetero- and homofermentative lactic acid bacteria. The effect of citrate on production of diacetyl and acetoin by four strains each of heterofermentative and homofermentative lactic acid bacteria capable of utilizing citrate was studied. Acetoin was quantitatively the more important compound. The heterofermentative bacteria produced no acetoin or diacetyl in the absence of citrate, and two strains produced traces of acetoin in its presence. Citrate stimulated the growth rate of the heterofermentative lactobacilli. Acidification of all heterofermentative cultures with citric acid resulted in acetoin production. Destruction of accumulated acetoin appeared to coincide with the disappearance of citrate. All homofermentative bacteria produced more acetoin and diacetyl in the presence of citrate than in its absence. Citrate utilization was begun immediately by the streptococci but was delayed until at least the middle of the exponential phase in the case of the lactobacilli.


Influence of specific growth rate on biomass yield, productivity, and compostion of Candida utilis in batch and continuous culture. Candida utilis was grown in batch and continuous culture on prickly pear juice as sole carbon and energy source. In batch culture the maximum specific growth rate (mum) and the substrate yield coefficient (Yps) varied according to sugar concentration. When the fermentation was carried out with 1% sugar, mum and Ys were 0.47/h and 42.6%, respectively. The best yields occurred in a chemostat at the pH range of 3.5 to 4.5 and temperature of 30 C. A beneficial effect on Ys was observed when the dilution rate (D) was increased. At a D of 0.55/h, the productivity was 2.38 g/liter per h. The maintenance coefficient attained a value of 0.09 g of sugar/g of biomass per h. Increases of D produced higher protein contents of the biomass. The information obtained indicates that protein production with Candida utilis, using prickly pear juice, should be carried out a high dilution rates where the Ys and protein content of the cell mass are also higher.


Chemical manipulation of the heat resistance of Clostridium botulinum spores. The chemical forms of Clostridium botulinum 62A and 213B were prepared, and their heat resistances were determined in several heating media, including some low-acid foods. The heat resistance of C. botulinum spores can be manipulated up and down by changing chemical forms between the resistant calcium form and the sensitive hydrogen form. The resistant chemical form of type B spores has about three times the classical PO4 resistance at 235 F (112.8 C). As measured in peas and asparagus, both types of C. botulinum spores came directly from the culture at only a small fraction of the potential heat resistance shown by the same spores when chemically converted to the resistant form. The resistant spore form of both types (62A and 213B), when present in a low-acid food, can be sensitized to heating at the normal pH of the food.


Poliovirus survival and movement in a sandy forest soil. Movement of poliovirus I (Chat) through nonsterile core samples of a sandy forest soil was monitored, using several regimens of loading with either dechlorinated final effluent from an operating activated sludge treatment plant or distilled water. Stimulated cycles of rainfall and effluent applications, resulting in ionic gradients, were shown to affect virus movement. Such studies indicate that poliovirus applied in effluents may move considerable distances through this soil after rainfall. Survival of poliovirus in the soil at 4 and 20 C has been monitored for 84 days. During this period, the capacity of the virus to migrate is unchanged.


Surface balance study of the interaction between microorganisms and lipid monolayer at the air/water interface. Using the surface balance technique, we have compared the interaction between Acholeplasma laidlawii and some marine bacteria towards different types of monolayered lipid films. Cells from A. laidlawii and Serratia marinorubra penetrate the film, whereas cells from Psuedomonas fluorescens form a  layer underneath the film. The forces that bind microorganisms to the air/water interface are not strong enough to scatter a condensed monolayer but increase the strength of loosely packed monolayers.


Toxicity of natural pyrethrins and five pyrethroids to fish. The toxicity of natural pyrethrins and five pyrethroids was determined with coho salmon (Oncorhynchus kisutch), steelhead trout (Salmo gairdneri), fathead minnow (Pimephales promelas), channel catfish (Icatlurus punctatus), bluegill (Lepomis macrochirus), and yellow perch (Perca flavescens). The 96-hour LC50's in static tests at 12 degrees C ranged from 24.6 to 114 mug/l of natural pyrethrins and from 0.110 to 1,140 mug/l of pyrethroids. Two pyrethroids, RU-11679 and SBP-1382 (R), were over 10 times more toxic than pyrethrum extract in the flow-through tests. Coldwater species of fish were more sensitive than warmwater species to all the compunds. Temperature (12-22 degrees C) influences the toxicity of natural pyrethrin and the pyrethroids. The natural pyrethrin was more toxic to fish in pH 6.5 than in pH 9.5 water, but the toxicity of pyrethroids was not influenced in the pH range. The two most toxic pyrethroids, RU-11679 and SBP-1382, Were deactivated more rapidly in water solutions than natural pyrethrins, S-bioallethrin, dimethrin, and d-trans allethrin.


Acute toxicity of selenium dioxide to freshwater fishes. Acute toxicity tests of selenium dioxide were conducted for 96 to 336 hr in intermittent-flow bioassay systems using six species of freshwater fish. The decreasing order of species sensitivity was: fathead minnow, flagfish, brook trout, channel catfish, goldfish, and bluegil. Curves relating median lethal concentration to exposure time for each species exposed for more than 168 hr were sigmoid in shape and were characterized by a change in slope indicating a more rapid mortality rate after 96 to 168 hr toxicant exposure. The 96-hr LC50 estimates ranged from 2.9 mg/L SeO2 for fathead minnow fry to 40.0 mg/L for bluegill juveniles. Effects of brief toxicant exposure (24 hr) on fathead minnow and flagfish juveniles included limited delayed mortality and no effects on growth over a 28-day period.


Effect of alpha and beta adrenergic blockade on epinephrine induced pulmonary insufficiency. Recent studies demonstrated that epinephrine causes significant pulmonary A-V shunting. This study reports the effect of alpha and beta adrenergic blockade on this shunting. Sixty-three anesthetized mongrel dogs were ventilated with a mechanical respirator. Measurements of (1) the pulmonary shunt, (2) cardiac output, (3) mean pulmonary artery, pulmonary capillary wedge and systemic pressures, and (4) pulmonary and systemic vascular resistances were obtained at 5, 15 and 30 minute intervals during the first hour and hourly for 5 hours. Fifteen dogs received no treatment. All others received epinephrine hydrochloride, 2 mug/kg/min for 5 hours. Ten received epinephrine only. Ten were pretreated with propranolol hydrochloride, 250 mug/kg, 12 with phenoxybenzamine, 1 mg/kg, and 16 with phenoxybenzamine and propranolol. Propranolol significantly decreased the epinephrine induced pulmonary shunt at all times and was the most effective drug. Phenoxybenzamine decreased the early shunting, but less than propranolol, and did not decrease the late shunting. Blockade with propranolol and phenoxybenzamine was less effective than propranolol alone. Based on the observed hemodynamic changes it was suggested that beta blockade is effective in reducing epinephrine induced pulmonary insufficiency by favorably altering the flow and distribution of pulmonary blood flow which in turn decreases epinephrine induced ventilation-perfusion inequalities and capillary hypertension both of which result in shunting. Conversely phenoxybenzamine has an unfavorable effect on the pulmonary flow. These studies support previous work in animals and man which showed that beta adrenergic stimulation is important in the pathogenesis of pulmonary insufficiency. Because the amounts of epinephrine used produce blood levels observed in critical illness, these studies add support to a relationship between the increased catecholamine stimulation of critical illness and the associated and often unexplained pulmonary insufficiency.


Revascularization of the heart through the coronary veins. Fifty-six dogs were used in a study to evaluate perfusion of the left anterior descending vein by the internal mammary artery in hearts with normal coronary arteries and those with ligated desending coronary arteries. Perfusion of the myocardium with arterial blood through the cardiac veins offers minimal immediate protection from infarction, as evidenced by light and electron microscopy studies. This protection is of short duration due to intimal fibrosis and luminal stenosis or obstruction of the perfused veins. Nineteen animals in which the coronary vein was perfused and the corresponding coronary artery was not ligated died within sixty hours from the time of operation. Pathological examination revealed patent grafts in all the animals. There was marked congestion of the myocardium with petechial hemorrhages over the surface of the heart. No evidence of myocardial infarction was found.


Dirofilaria immitis (dog heartworm) as a pulmonary lesion in humans. Dirofilaria immitis, the dog heartworm, has been identified in the pulmonary granulomas of 5 patients from the greater Charleston area; this is the largest series of such cases from one medical center. The patients had no pulmonary symptoms. On roentgenogram the lesions were all about 2 cm in size, of uniform light opacity, and located near the pleural surface. Thoractomy was performed in each instance because of the possiblity of carcinoma. The association of granuloma formation, pulmonary infarct, and eosinophilic infiltration led to the suspicion of dirofilaria, which was confirmed in each case.


Benorylate interaction with indomethacin and phenylbutazone. The simlutaneous oral administration of benorylate (4-(acetamido) phenyl 2-acetoxybenzoate) with either indomethacin or phenylbutazone to rats suffering from Freund's adjuvant-induced arthritis leads to an anti-inflammatory effect which is significantly greater than the effect of the same drugs administered alone. Such an additive anti-inflammatory effect is not apparent when the metabolites of benorylate (paracetamol, acetylsalicylic acid) are administered with indomethacin or phenylbutazone. Paracetamol does not increase the anti-inflammatory effect of indomethacin or phenylbutazone and acetylsalicylic acid clearly antagonizes it. The molecule of benorylate itsel is therefore responsible for the additive anti-inflammatory effect. However, if antipyretic activity (yeast-induced hyperthermia) is examined instead of anti-inflammatory activity, the simultaneous oral administration of the different drugs always produces an additive effect. It is concluded that the antagonism between indomethacin or phenylbutazone and non-steroidal anti-inflammatory drugs other than benorylate is present at some receptors but not all. The clinical implications of the results are discussed.


A controlled trial of amantadine in drug-induced extrapyramidal disorders. Presently marketed antiparkinsonism drugs are potent anticholinergic agents that, while effective in treating extrapyramidal symptoms (EPS), also are productive of or can exacerbate a number of side effects associated with psychotropic drugs. Some of these include gastrointestinal disturbances, visual difficulties, and tardive dyskinesia. A double-blind study was carried out to assess the efficacy (and adverse effects) of amantadine hydrochloride--an agent without appreciable anticholinergic activity--for the treatment of drug-induced EPS. Amantadine was found to be comparable in effect to benztropine mesylate, but with fewer side effects. The potential role of amantadine may be in the treatment of patients with drug-induced EPS for whom medication with anticholinergic properties is contraindicated.


Metabolism and ultrastructure in ovaries of alloxan-diabetic juvenile rats. Tests were carried out on the influence of alloxan-induced diabetes mellitus on the metabolism and the ultrastructure of ovaries of juvenile rats. The diabetes mellitus caused the following changes in the metabolism: reduction in the concentration of ATP and NADPH, increase in the lactate/pyruvate quotient to above 40, reduction in the ATP/ADP quotient to below 1, reduction in the level of activity of the hydrogen-conveying enzymes G-6-P-dehydrogenase, isocitrate dehydrogenase and malate dehydrogenase, increase in the level of activity of the alkaline phosphatase, reduction of the protein content. Ultrastructure: almost complete disappearance of the rough endoplasmic reticulum, shrinkage of the mitochondria, reduction of the cristae and condensation of the matrix. The smooth endoplasmic reticulum remains unchanged, the extent of the Golgi-complex is reduced. Easy removal of the lipid deposits.


Human tear pH. Diurnal variations.

Human tear pH. Diurnal variations. Using a closed chamber microelectrode system, the tear pH levels of sixteen subjects were monitored throughout the waking extent of five days. In addition to the absolute mean pH differences found among subjects, diurnal patterns of pH change could be identified for the majority. The amplitudes and periods of these cycloid patterns, however, were distinctive to each individual. Also, tear pH levels following periods of prolonged eye closure were found to be notably more acid than those associated with the waking hours.


Reactivity and ionization constants of the lysine residues in apovitellenin i of emu egg yolk low-density lipoprotein by competitive labelling. Competitive labelling with[14C]acetic anhydride over a range of pH values has been used to explore the surface topography of the apovitellenin I moiety in emu egg yolk low-density lipoprotein. The reaction of the lysine  xi-amino groups with acetic anhydride has been related to pH in a set of titration curves; from these, the reactivities relative to alanine and the ionization constants of all but the amino terminal lysines have been determined. All lysines have near normal pKa values around 10, and lower than normal reactivities (except the amino terminal lysine). At pH values above 10, the titration curves show breaks where the epsilon-amino groups become much more reactive, except for lysine 71 which in this regard behaves like a normally ionizing lysine in not showing a discontinuity. Most of the basic residues in this apoprotein may occur clustered at the surface of the molecule. This accounts best for the observed low reactivities and pKa values. The amino terminal lysine residue is presumably completely exposed to the aqueous environment.


Properties and specificity of a second metal chelator-sensitive proteinase in the keratinolytic larvae of the webbing clothes moth. The properties of a second metal chelator-sensitive proteinase (metalloproteinase 2) from the larvae of the webbing clothes moth, Tineola bisselliella, have been studied. The pH optimum for casein digestion was 9-4 and the enzyme showed high stability between pH 8 and 11, but very poor stability at acid pH. The proteinase was inhibited by EDTA, but not by an EDTA-calcium complex. EDTA inhibition could be reversed by addition of a slight excess of calcium or zinc ions. The cleavage specificity of metalloproteinase 2 against the A and B chains of S-carboxymethyl insulin was almost identical to that found previously for metalloproteinase 1.


Aminopeptidases in webbing clothes moth larvae. properties and specificities of enzymes of highest electrophoretic mobility. The group of aminopeptidase bands from Tineola bisselliella larvae with highest electrophoretic mobility in polyacrylamide gels were purified further and partially separated by ion exchange chromatography. Three aminopeptidase bands were present in this material and were very similar with respect to their pH optima (7-7), their molecular weight of 94,000, their responses to metal ions and enzyme inhibitors and in their substrate specificity requirements. Kinetic constants were obtained for the hydrolysis of 17 different alpha-aminoacyl-beta-naphthylamides by these aminopeptidases, the most favoured substrates being the derivatives of alanine, methionine, proline, leucine, glycine, glutamic acid, lysine and arginine. The enzymes also hydrolyse amino acid amides, dipeptides, dipeptide amides, tripeptides and oligopeptides at the N-terminal end. These enzymes differ from the other aminopeptides in T. bisselliella in being able to hydrolyse bonds involving proline.


Anti-leukaemia activity as a bystander effect of graft-versus-host reactions. The production of graft-versus-host (GVH) reactions in (PVGc X Wistar) F1 hybrids by the transfer of PVGc spleen cells resulted in significant resistance of these recipients to a subsequent challenge with the PVGc leukaemia. Protection was markedly dependent on dose and timing of allogeneic cell transfer and was abrogated by irradiation of the cells prior to transfer. GVH activity was shown to be a prerequisite for induction of the protective effect but was equally effective when produced by the transfer of Wistar spleen cells in place of PVGc cells. These points, plus the fact that invitro investigations of possible immune mechanisms failed to demonstrate cytotoxic immunity in treated rats, suggested a nonspecific "bystander" effect as the mechanism of protection. The implications of such a mechanism are discussed.


The effect of aflatoxins on the incorporation of RNA and protein precursors by isolated hepatocytes. Hepatocytes prepared by a simplified enzymatic technique were active in the incorporation of RNA and protein precursors into acid-insoluble material. The incorporation of RNA precursors was very markedly inhibited by low levels of aflatoxin B1 and G1 but not by aflatoxins B2 and G2. The activity of mixed function oxidases (MFO), the drug-metabolizing system of the endoplasmic reticulum, could be suppressed in these cells by SKF525A or stimulated by NADPH. SKF525A caused a reduction in the inhibition by aflatoxin B1 of the incorporation of RNA precursor into macromolecules. This finding suggests that a metabolite of aflatoxin B1 is the actual inhibitor of RNA synthesis in the cells. Measurement of lactate dehydrogenase activity showed these cells to be leaky on incubation at 37 degrees C and thus not suitable for studies of protein secretion.


Passive serum sickness in the mouse: the role of vasoactive amines in glomerular deposition of immune complexes. This paper reports a study of the importance of vasoactive amines in glomerular localization of passively administered immune complexes in the mouse. Two strains of mice were investigated, one sensitive and the other relatively resistant to the anaphylactogenic effect of intravenously administered immune complexes. The effect on glomerular deposition immune complexes in animals of both strains treated with either vasoactive amine depletors or antagonists leads to the conclusion that in this experimental system vasoactive amines do not play a major part in the glomerular localization of immune complexes.


Intensive care of the fetus in breech labour. A review of 186 cases of breech presentation with a corrected perinatal mortality rate of 0-54 per cent is presented. Details of paediatric follow-up are given. Careful selection of patients for vaginal delivery and the liberal use of Caesarean section are advocated. The importance of asphyxia as the main danger of breech delivery is emphasized and the use of fetal blood sampling as a practicable method of detecting early asphyxia is discussed.


The effects of grass and concentrate diets on the specific activities of some enzymes of hepatic carbohydrate metabolism in sheep. Feeding sheep a concentrate diet compared with grass diets increased the hepatic specific activities of the three glycolytic enzymes studied, and that of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and reduced the specific activity of D-fructose-I, 6-diphosphate I-phosphohydrolase (EC 3.1.3.11). The specific activities of phosphogluconate dehydrogenase (EC 1.1.1.43) and malate dehydrogenase (decarboxylating) (NADP) (EC 1.1.1.40) were unaffected by diet.


Light-induced glutamate transport in Halobacterium halobium envelope vesicles. II. Evidence that the driving force is a light-dependent sodium gradient. Illumination of cell envelope vesicles from H. halobium causes the development of protonmotive force and energizes the uphill transport of glutamate. Although the uncoupler, p-trifluoromethoxycarbonyl cyanide phenylhydrazone (FCCP), and the membrane-permeant cation, triphenylmethylphosphonium (TPMP+), are inhibitory to the effect of light, the time course and kinetics of the production of the energized state for transport, and its rate of decay after illumination, are inconsistent with the idea that glutamate accumulation is driven directly by the protonmotive force. Similarities between the light-induced transport and the Na+-gradient-induced transport of glutamate in these vesicles suggest that the energized state for the amino acid uptake in both cases consists of a transmembrane Na+ gradient (Na+out/Na+in greater than 1). Rapid efflux of 22Na from the envelope vesicles is induced by illumination. FCCP and TPMP+ inhibit the light-induced efflux of Na+ but accelerate the post-illumination relaxation of the Na+ gradient created, suggesting electrogenic antiport of Na+ with another cation, or electrogenic symport with an anion. The light-induced protonmotive force in the H. halobium cell envelope vesicles is thus coupled to Na+ efflux and thereby indirectly to glutamate uptake as well.


Physical studies of the nonhistone chromosomal proteins HMG-U and HMG-2. The nonhistone chromosomal proteins, HMG-1 and HMG-2, have a folded conformation, with a high alpha-helical content, over a wide pH range. At high and low pH values, the molecules unfold. Both molecules contain cysteine and tryptophan. The tryptophans appear to be buried in the folded form. HMG-1 shows aggregation at pH 5.7, as does HMG-2 at pH 9.0. The folded form is insensitive to high concentrations of salt, suggesting that charge-charge interaction plays no role in stabilizing the tertiary structure.


Synthesis and properties of carbonylbis(methionyl)insulin, a proinsulin analogue which is convertible to insulin by cyanogen bromide cleavage. The preparation and use of carbonylbis (L-methionine p-nitrophenyl ester) as a reversible cross-linking reagent for insulin are described. The reaction of 1 equiv of reagent with zinc insulin in dimethylformamide in the presence of triethylamine yields as one of the products NalphaA1, NepsilonB29-carbonylbis(methionyl)insulin, (CBM-insulin). The CBM-insulin was characterized by end group analysis and by the products formed on tryptic and chymotryptic cleavage. It possessed 91% of the immunological and 6.5% of the hormonal activity of insulin. Treatment of CBM-insulin with cyanogen bromide (CNBr) in 70% formic acid for 1 h resulted in nearly complete removal of the methionine bridge to yield insulin. A small amount of a side product was removed on DEAE-cellulose at pH 7.2 to give an overall recovery of insulin of 70-80%. Oxidative sulfitolyses of CBM-insulin gave the hexa(S-sulfonate) which was reduced with dithiothreitol to yield reduced CBM-insulin. The latter compound, containing 6 sulfhydryls, exhibited a pH-dependent circular dichroic spectrum. The form at pH 10 exhibited a spectrum typical of random coil which was converted to a form at pH 7.8 which was characterized by a negative extremum at 213 nm. The change in the spectrum at 213 nm with pH was characterized by an apparent pKa of 8.5. Studies on the reoxidation of reduced CBM-insulin were performed at pH values between 7.8 and 10 and at protein concentrations of 0.01-1 mg/ml. The best yields (ca. 85%) of the correctly paired disulfide bonds were obtained in reoxidations at pH 9.5-10 at protein concentration of 0.01-0.1 mg/ml. CBM-insulin, which had been isolated from reoxidation at high pH of the reduced CBM-insulin, was cleaved by CNBr to yield a fully active insulin in an overall yield of 60% from the reduced CBM-insulin.


DNA-dependent RNA polymerase from the thermophilic bacterium Caldariella acidophila. Purification and basic properties of the enzyme. A DNA-dependent RNA polymerase has been isolated from Caldariella acidophila, a thermophilic bacterium living in acidic hot springs at temperatures ranging from 63 to 89 degrees C. The enzyme was purified 180-fold and is composed of five different subunits having the following molecular weights: a = 127000, b = 120000, c = 72000, d = 65000, and e = 38000. The enzyme is activated by Mn2+ and Mg2+ and exhibits optimal activity in the presence of 0.5 mM Mn2+. The activity depends on ionic strength, with a maximum at 0.25 M KCl, and exhibits a pH optimum at 7.8 in the presence of Tris-HCl buffer. The enzyme shows a high degree of thermophilicity, its temperature optimum being 80 degrees C in the in vitro assay. The thermophilicity of C. acidophila RNA polymerase allows studies on enzyme-template interactions to be performed in a temperature range where many templates are close to their Tm.


Induction of stable protein-deoxyribonucleic acid adducts in Chinese hamster cell chromatin by ultraviolet light. Ultraviolet (uv)-light-mediated formation of protein-DNA adducts in Chinese hamster cell chromatin was investigated in an attempt to compare chromatin alterations induced in vitro with those observed in vivo. Three independent methods of analysis indicated stable protein-DNA associations: (1) a membrane filter assay which retained DNA on the filter in the presence of high salt-detergent; (2) a Sepharose 4B column assay in which protein eluted cincident with DNA; and (3) a CsCl density gradient equilibrium assay which showed both protein and DNA banding at densities other than their respective native densities. Treatment of the irradiated chromatin with DNase provided further evidence that protein-DNA and not protein-protein adducts were being observed in the column assay. There is a fluence-dependent response of protein-DNA adduct formation when the chromatin is irradiated at low ionic strength and is linear for protein over the range studied. When the chromatin is exposed to differing conditions of pH, ionic strength, or divalent metal ion concentration, the quantity of adduct formed upon uv irradiation varies. Susceptibility to adduct formation can be partially explained in terms of the condensation state of the chromatin and other factors such as rearrangement, denaturation, and dissociation of the chromatin components. Besides providing information on the biological significance of these types of uv-induced lesions, this technique may be useful as a probe of chromatin structure.


D-Mannitol dehydrogenase from Absidia glauca. Purification, metabolic role, and subunit interactions. When Absidia glauca was grown in minimal media with D-mannitol as the only source of carbon, an NAD+ specific D-mannitol dehydrogenase (EC 1.1.1.67) was induced. The crude extract also gave evidence of mannitol kinase, mannitol-1-phosphate dehydrogenase, phosphofructokinase, and L-iditol dehydrogenase activity. The heat labile purified preparation was judged enzymically homogeneous based on evidence derived from substrate specificity studies and activity staining, following disc gel electrophoresis. The enzymic monomer, with a weight of about 67000 daltons, slowly polymerizes when stored at -20 degrees C, giving a multiplicity of protein bands on electrophoresis distributed predominantly across a spectrum from dimer to pentamer, with enzymic activity resident predominantly in even multiples of the monomer. Depolymerization occurred rapidly (hours) when a frozen preparation was brought to and held between 4 and 20 degrees C. Aggregate fragmentation with sodium dodecyl sulfate showed a time-temperature dependence, terminating in a subunit component of 13000 daltons. pH optimum for polyol oxidation occurs at 9.6 (NaOH-glycine buffer) while ketose reduction proceeded most rapidly at pH 7.0-7.2 (phosphate buffer). A regulatory role is suggested for this enzyme based on dead-end inhibition by mannitol 1-phosphate, multiple enzyme forms, and its locus at the initiation site for mannitol utilization. The physiological relevance of low-temperature aggregation to regulatory control remains to be established.


Glutamine synthetase of Bacillus stearothermophilus. Regulation, site interactions, and functional information. The action of various feedback modifiers on Bacillus stearothermophilus glutamine synthetase has been investigated by initial velocity kinetics, using the Mn2+-stimulated biosynthetic assay at 55 degrees C. The most potent inhibitors, used singly, are AMP, L-glutamine, and L-alanine. Other modifiers of significance include glycine, CTP, L-histidine, glucosamine 6-phosphate, and GDP. Marked synergism of action is observed for AMP in the presence of L-glutamine, L-histidine, ADP, or glucosamine 6-phosphate (glucosamine-6-P), and for CTP with ADP or GDP. Inhibition by saturating levels of many modifiers is either less than 100%, or is not overcome by elevated substrate levels, or both. This argues for modifier binding sites separate from substrate sites, notably in the cases of AMP, L-glutamine, glycine, L-alanine, glucosamine-6-P, and CTP. Glycine and L-alanine are Vmax inhibitors, whereas L-glutamine, glucosamine-6-P, GDP, and CTP alter the binding of L-glutamate. ADP and L-histidine apparently can compete directly with MnATP, but AMP alters Mn-ATP binding from a separate site. The action of several modifiers requires or is enhanced by bound substrates. Considerable antagonistic interaction is observed in experiments with modifier pairs, but the most potent inhibitors show synergistic or cumulative (independent) interactions. One may interpret antagonistic effects as due to (a) overlapping modifier domains, or (b) separate but antagonistically interacting sites. Either interpretation leads to a scheme for modifier-substrate and modifier-modifier site interactions in which the thermophilic enzyme must maintain and stabilize a great deal of complex functional information under extreme environmental conditions.