Concentration of poliovirus from tap water using positively charged microporous filters. Microporous filters that are more electropositive than the negatively charged filters currently used for virus concentrations from water by filter adsorption-elution methods were evaluated for poliovirus recovery from tap water. Zeta Plus filters composed of diatomaceous earth-cellulose-"charge-modified" resin mixtures and having a net positive charge of up to pH 5 to 6 efficiently adsorbed poliovirus from tap water at ambient pH levels 7.0 to 7.5 without added multivalent cation salts. The adsorbed virus were eluted with glycine-NaOH, pH 9.5 to 11.5. Electropositive asbestos-cellulose filters efficiently adsorbed poliovirus from tap water without added multivalent cation salts between pH 3.5 and 9.0, and the absorbed viruses could be eluted with 3% beef extract, pH 9, but not with pH 9.5 to 11.5 glycine-NaOH. Under water quality conditions in which poliovirus recoveries from large volumes of water were less than 5% with conventional negatively charged filters and standard methods, recoveries with Zeta Plus filters averaged 64 and 22.5% for one- and two-stage concentration procedures, respectively. Electropositive filters appear to offer distinct advantages over conventional negatively charged filters for concentrating enteric viruses from water, and their behavior tends to confirm the importance of electrostatic forces in virus recovery from water by microporous filter adsorption-elution methods.


Development of a quantitative method for the detection of enteroviruses in soil. A method is described for efficiently concentrating enteroviruses from soil. Viruses were eluted from soil by mechanical agitation in high pH glycine buffer containing ethylenediaminetetraacetic acid. The eluted viruses were concentrated on a floc that formed de novo upon adjustment of the soil eluate to 0.06 M aluminum chloride and pH 3.5. Viruses not pelleted with the floc were concentrated by adsorption to and elution from membrane filters. This method yielded an average efficiency of 66% recovery from loamy sand soil for four enteroviruses. Virus recovery from soil was consistently high, with samples ranging in size from 25 to 500 g. The method was used successfully to isolate naturally occurring viruses from soil beneath a wastewater land treatment site. Recovery of enteroviruses by this method form different types of soil was dependent on percentage of clay, surface area, and cation exchange capacity. Recovery was not dependent on soil saturation pH or on percentage of organic matter. This method should prove useful for studying enterovirus migration and survival during the land application of domestic sewage.


Determinants in microbial colonization of the murine gastrointestinal tract: pH, temperature, and energy-yielding metabolism of Torulopsis pintolopesii. Torulopsis pintolopesii is an indigenous yeast that colonizes the secreting epithelia in the stomachs of mice and rats. A wild-type strain of this microbe was isolated and identified. To attempt to learn characteristics of the yeast that are advantageous to it in colonizing its natural habitat in vivo, we examined some aspects of its nutrition and energy-yielding metabolism and some environmental conditions that influence its growth in vitro. The yeast appeared to be limited in the compounds it can utilize as carbon and nitrogen sources. It grew best at 37 degrees C and did not grow at 23 or 43 degrees C. It grew optimally at neutral pH but could grow aerobically at pH values as low as 2.0 and anaerobically at pH values as low as 3.4. As assessed by measurements of growth rates and yield coefficients, it grew better aerobically than anaerobically. When grown aerobically, it had a cyanide-sensitive system for taking up O(2) and tested positively for cytochrome c oxidase activity. A petite mutant strain isolated from the wild-type strain had a growth rate and yield coefficient when incubated aerobically that were essentially the same as those of the wild-type parent grown anaerobically. Likewise similar to the wild-type parent grown anaerobically, the petite strain, though incubated aerobically, did not take up O(2). Yeast-free mice associated with either the wild-type or the petite mutant strain were colonized at essentially the same rates and to similar final population levels by both strains. The yeast's capacity to respire may be of little advantage to it in its natural environment. By contrast, its abilities to grow best at 37 degrees C and to grow at low pH values are undoubtedly advantageous characteristics in this respect. The limitations in its carbon and nitrogen nutrition are difficult to evaluate as ecological factors in its colonization of the natural habitat.


Measurement of the inactivation kinetics of poliovirus by ozone in a fast-flow mixer. Inactivation kinetics of poliovirus type 1 in ozone demand-free water was investigated by utilizing a fast-flow mixing apparatus. Ozonated water and a solution of ozone demand-free water containing a known quantity of poliovirus type 1 were introduced simultaneously into a mixing chamber, both at a constant rate. This mixture was then passed through a narrow tube of known length and diameter into a neutralizing solution. By altering the rate of introduction and/or tube length, different contact periods between ozone and virus could be determined with an accuracy of 0.01 s. Inactivation of the poliovirus occurred in two steps. During the first step, which lasted for 0.2 to 1.0 s, 95 to 99% of the virus was inactivated, depending on the ozone concentration (which ranged from 0.1 to 2.0 mg/liter). The second step apparently continued for several minutes; in this period the remainder of the virus was inactivated. An obvious dose-response relationship was demonstrated during the first step of the inactivation curve. The pH of the water slightly affected the viral inactivation rate, but these small differences seem to have no practical value.


Effect of environmental parameters on the biodegradation of oil sludge. A laboratory study was conducted with the aim of evaluating and optimizing the environmental parameters of "landfarming", i.e., the disposal by biodegradation in soil of oily sludges generated in the refining of crude oil and related operations. Oil sludge biodegradation was monitored by CO2 evolution and by periodic analysis of residual hydrocarbons. The parameters studied were soil moisture, pH, mineral nutrients, micronutrients, organic supplements, treatment rate, teratment frequency, and incubation temperature. Oil sludge biodegradation was optimal at a soil water-holding capacity of 30 to 90%, a pH of 7.5 to 7.8, C:N and C:P ratios of 60:1 and 800:1, respectively, and a temperature of 20 degrees C or above. Addition of micronutrients and organic supplements was not beneficial; sewage sludge interfered with hydrocarbon biodegradation. Breakdown of the saturated hydrocarbon (alkane and cycloalkane) fraction was the highest at low application rates, but higher application rates favored the biodegradation of the aromatic and asphaltic fractions. An application rate of 5% (wt/wt) oil sludge hydrocarbon to the soil (100,000 liters/hectare) achieved a good compromise between high biodegradation rates and efficient land use and resulted in the best overall biodegradation rate of all hydrocarbon classes. Frequent small applications resulted in higher biodegradation than single large applications. Two 100,000-liter/hectare (255 barrels per acre) or four 50,000-liter/hectare oil sludge hydrocarbon applications per growing season seem appropriate for most temperate zone disposal sites.


Magnitude of pollution indicator organisms in rural potable water. A total of 460 water samples were randomly drawn from the potable water supply sources of rural communities in three counties of South Carolina. About 10% of the population, not incorporated in municipalities, was sampled. The samples were tested for total coliforms, Escherichia coli, and fecal streptococci. Significant levels of these pollution indicator organisms were detected in almost all the water supplies. Total coliforms were the most common, and only 7.5% of the water supplies were uncontaminated. E. coli, considered a reliable indicator of recent and dangerous pollution, was observed in 43% of the water supplies. Statistical analyses indicated that the bacterial populations, especially E. coli, were associated with the supply source depth and its distance from the septic tank. Total coliform counts were also weakly correlated to the pH of the water.


Partial reactivation of chlorine-treated echovirus. After treatment of a dispersed suspension of echovirus with HOCl, much of the lost plaque titer was restored if the treated virus was induced to aggregate by adjustment of the suspending medium to pH 4.5. This did not appear to be a repair of individual virions but rather a special kind of multiplicity-related increase in plaquing efficiency which occurred when the host cell received several of the damaged virions in a clump.


Graft patency in coronary bypass surgery. A total of 514 vein bypass grafts and 49 internal mammary (IMA) grafts in 328 patients were studied after operation. Forty-two vein bypass grafts were performed without the use of a pump oxygenator, with a patency rate of 52%. When a pump oxygenator was used, the patency rate for vein bypass grafts was 78%. Patency rates for IMA grafts were 70% and 86%, respectively. In a small group of patients, endarterectomy with vein bypass grafts resulted in a patency rate of 59% in the right coronary artery, 88% in the left anterior descending coronary artery, and 74% in the circumflex artery. Except for the right coronary artery, these results compare favorably with those from vein bypass graft patency without endarterectomy. On the basis of these findings, insertion of bypass grafts into the coronary arteries without the use of a pump oxygenator cannot be recommended, unless the technique employed can be shown to produce graft patency rates comparable to those resulting from grafts done with the use of a pump oxygenator. Endarterectomy to the left anterior descending and circumflex arteries would not appear to affect vein bypass graft patency.


Respiratory acidosis with the small Storz-Hopkins bronchoscopes: occurrence and management. Carbon dioxide retention in the Storz rigid ventilating bronchoscope with the Hopkins lens system was investigated in the laboratory. The 3.5, 4.0, and 5.0 30-cm Storz bronchoscopes with a 3.95-mm (outside diameter) telescope lens were used in 10 mongrel dogs weighing between 8 and 15 kg. Significant (p less than 0.01) accumulation of arterial carbon dioxide tension (PaCO2) (respiratory acidosis) was observed after 5 and 10 minutes of ventilation through the 3.5 and 4.0 bronchoscopes, but no significant increase in PaCO2 was noted with the 5.0 bronchoscope. There was no significant change in arterial oxygen tension under the same conditions. Manual compression of the upper anterior abdominal wall during expiration was applied during bronchoscopy in 6 children. Arterial blood samples were taken before insertion of the bronchoscope and 5 minutes later with and without abdominal compression during expiration. A significant increase (p less than 0.05) in PaCO2 and a decrease in pH were observed after 5 minutes of the bronchoscopic procedure without manual compression of the abdominal wall, while no significant changes in PaCO2 were observed with abdominal compression.


Takayasu's arteritis: surgical considerations. Takayasu's arteritis, the nonspecific occlusive disease originally believed to affect young Oriental women exclusively, was identified in a 19-year-old man who had severe involvement of the aortic arch and its branches. Successful surgical treatment was accomplished with a bypass from the ascending aorta to the supraceliac abdominal aorta using a tube graft. The right vertebral artery, as the single patent vessel reaching the head, was revascularized distal to the stenosis with a tube graft that extended from the aortic graft. Revascularization of ischemic organ systems and body areas may be accomplished in most patients with Takayasu's arteritis using the bypass concept and fabric conduit grafts.


ECG changes during long-term minoxidil therapy for severe hypertension. A prospective assessment of the effects of minoxidil on the ECG was carried out in a series of 80 patients with severe hypertension, representing an observation of 139.6 patient years. In combination with beta-adrenergic blocking agents and diuretics, minoxidil resulted in substantial reductions in blood pressure at rest and during exercise. Concomitant with the initiation of minoxidil treatment, ECG changes consisting of flattening or inversion of the T waves, ranging from slight to very marked, were observed in 90% of the patients. While these changes may be potentially disconcerting, the observations of this study show that they are not related to changes in heart rate or other clinical criteria associated with myocardial ischemia. They are not influenced by exercise or beta-blockade and they generally revert to control appearance during chronic treatment, at which time accompanying improvement in preexisting abnormalities of the T wave was frequently seen. During long-term treatment, a substantial reduction of increased QRS voltages was observed.


Lateral cerebral ventricular enlargement in chronic schizophrenia. To investigate if cerebral ventricular enlargement is associated with chronic schizophrenia, computerized tomography scans from 73 psychiatric patients were compared with 56 asymptomatic volunteers all less than 50 years old. Ventricular size was significantly greater in the subgroup of 58 chronic schizophrenic patients than in the controls. Of the chronic schizophrenic patients, 40% were outside the control range; 53% exceeded 2 SDs of the control mean. Neither duration of illness nor length of hospitalization correlated with ventricular size. The 44 chronic schizophrenic patients who had never been treated with electroshock therapy (EST) had larger ventricles than controls. A group of seven nonchronic schizophrenic patients also had enlarged ventricles; the eight patients who were either schizoaffective or nonschizophrenic did not differ from controls. This study shows that lateral cerebral ventricular enlargement is associated with chronic schizophrenia; it suggests that this is not a result of treatment.


NIMH clinical research branch collaborative program on the psychobiology of depression. This is a report on the history and implications of the collaborative effort that evolved from the 1969 National Institute of Mental Health conference on the psychobiology of depression. The major issues identified at that time were the need to (1) assess relative validities of current systems of nosology and (2) retest critical biological hypotheses concerning the etiology and nature of the depressive disorders. Research was required that would be multidisciplinary and involve clinical settings treating diverse types of depression. The objectives and the nature of the biological and clinical collaborative programs that were designed to address these problems are described. These unique programs, initiated in the early 1970s, currently span research on nosology, genetics, neurochemistry, neuroendocrinology, and psychosocial factors. Although these studies are still in the early stages, they have resulted in significant methodologic developments in diagnosis, descriptive psychopathology, and biological measurements.


Clinical usefulness of sodium amobarbital interviewing. We report a double-blind, randomized, placebo-controlled study utilizing a within-subjects design on 20 hospitalized, psychiatric patients who participated in sodium amobarbital interviews to determine if the drug has a specific effect in eliciting clinically useful information. The patients selected had difficulty communicating with their primary therapists during the postadmission, diagnostic interviews. Two raters completed a Hamilton Depression Scale, a New Haven Schizophrenia Index, and a Brief Psychiatric Rating Scale after each interview. Although both the amobarbital and saline interviews were moderately useful in obtaining new information, we found no significant difference in the primary therapists' assessments of clinical usefulness. In addition, the drug interview did not uncover material that would aid in the differential diagnosis between depression and schizophrenia. There was, however, a significant negative correlation between the assessment of general usefulness and the time interval between admission and interviewing. We report our only exception, a case of catatonic schizophrenia, in which the patient responded specifically to the drug.


pH-induced platelet ultrastructural alterations. A possible mechanism for impaired platelet aggregation. After the observation that lavages with alkaline solutions exert a beneficial effect on the bleeding tendency and increase platelet aggregation in patients with intragastric hemorrhage, studies were undertaken to find a relationship between changes of pH and platelet morphology. Transmission and scanning electron microscopy showed that at a pH lower than 7.4, normal human platelets change their internal structure as well as their shape, becoming spheres deprived of pseudopodia. On the other hand, a pH higher than 7.4 induces transformation of platelet internal organelles similar to that caused by thrombin. At an alkaline pH, the platelets develop multiple pseudopodia that facilitate their attachment to each other. These findings may explain the increased platelet aggregation in alkaline medium demonstrated in one of our previous works.


The gastroesophageal scintiscan. Comparison of methods to demonstrate gastroesophageal reflux. The gastroesophageal (GE) scintiscan method of detecting GE reflux was evaluated in a series of 54 patients. Twenty-nine patients had evidence of GE reflux by fluoroscopically monitored barium swallow, esophagogastroscopy, or esophageal biopsy. The pH reflux study showed pathological reflux in four patients. The GE scintiscan demonstrated reflux in only four of these 29 patients. Although the GE scintiscan is a safe, noninvasive technique, in our experience, GE reflux was demonstrated only in those patients in whom reflux was easily verified by barium swallow, esophagoscopy, or esophageal mucosal biopsy. This study indicates that the GE scintiscan, like the pH reflux study, is too insersitive to be of consistent value in the diagnosis of GE reflux.


[Mediator stage in the functioning of the autonomic nervous system in pre- and postnatal ontogeny and the importance of its study for clinical application]. In 1972 a new definition "mediatory stage" in the development of the vegetative nervous system during ontogenesis was introduced and it was stated that at the prenatal stage in the heart, along the course of magistral vessels, and into other organs begin to sprout up at first nervous truncs lacking in mediators, and then, beginning from 8--9 weeks (in man), the mediators appear at first in cholinergic and then in adrenergic plexus (V. N. Shvalev et al., 1972). In clinical embryonal mortality is especially great during the stage of premediatory development, that corresponds to the period of placentation. The beginning of the mediatory stage (the 3d month of prenatal life) is characterized by an intensive differentiation of the neural apparatus in the heart and its innervated structures. At first cholinergic and then adrenergic nerve plexus are formed, nevertheless, by the time of birth these plexus, especially adrenergic ones are not yet fully differentiated. During the first year of life there is noted a rather high rate of so called "sudden death". During the following 3--4 years there is a new intensive rise in differentiation of the cardiac cholinergic and adrenergic nerve plexus and by the 6--8th year of age the density of these plexus reaches its maximum (about 10%). This index is constant up to 40 years of age, and then a noticeable decrease in the density of adrenergic plexus takes place, while that of cholinergic plexus remains rather constant. After 60 years of age an involution takes place at first in adrenergic and then in cholinergic plexus, and the "postmediatory" stage occurs. The problem of "sudden death" is closely connected with the changes described for the mediators in the cardiac and vascular plexuses (V. N. Shvalev, R. A. Stropus and E. K. Morozov, 1978).


Granulomatous angiitis of the CNS.

Granulomatous angiitis of the CNS. A 25-year-old man had granulomatous angiitis of the CNS. The disease began with symptoms of an upper respiratory tract infection; it had a relentless course, simulating viral encephalitis, with the patient dying some six months after the onset of symptoms. The lesions were confined to small intracranial arteries and veins, predominantly about the cerebellum. An unusual feature was the occurrence of a small (2 mm) aneurysm on a leptomeningeal artery deep in a cerebellar sulcus. There is a need for pooling data and pathological material by an international body to enable detailed expert analysis of large numbers of cases of cerebral angiitis that do not now constitute a homogeneous group.


Otitis media with effusion. Cytological and microbiological correlates. A quantitative cytologic study of 83 middle ear effusions from children with otitis media with effusion (OME) was made with the use of cytocentrifuge-prepared smears and was correlated with bacteriological findings. The identification of the types of cell present in an effusion was confirmed by electron microscopic findings. The most common cell types were neutrophilic leukocytes, lymphocytes, and macrophages, but a few eosinophils were found in only one case. The cytologic profile of an effusion correlated well with the bacterial culture results. Neutrophil counts were significantly higher in effusions that contained pathogenic bacteria (P less than .01) than in culture-negative effusions. Lymphocytes were the dominant cell type in culture-negative effusions. The results support the idea that bacteria are involved in the pathogenesis of many cases of OME.


Verapamil in the management of hypertensive patients. The hypotensive action of verapamil given orally was assessed in the outpatient management of patients with raised blood pressure, in a single blind study. Twenty-six hypertensive patients on no therapy, 14 on beta blockers and eight on methyldopa were more satisfactorily controlled when verapamil was substituted for the initial drug. There was a marked reduction in the number of side-effects, and the drug was given in many situations where beta blockade was contra-indicated.


Inborn errors of cobalamin metabolism: effect of cobalamin supplementation in culture on methylmalonyl CoA mutase activity in normal and mutant human fibroblasts. We have examined the effect of addition of hydroxocobalamin to growth medium on the activity of the adenosylcobalamin-requiring enzyme methylmalonyl CoA mutase in normal human fibroblasts and in mutant human fibroblasts derived from patients with inherited methylmalonicacidemia. The mutant cell lines were assigned to four distinct genetic complementation groups (cbl A, cbl B, cbl C, and cbl D), each deficient in some step in the synthesis of adenosylcobalamin from hydroxocobalamin. After control cells were grown in cobalamin-supplemented medium, mutase holoenzyme activitiy increased markedly in a time- and concentration-dependent fashion. Growth in cobalamin-supplemented medium had no effect on mutase activity in some mutant lines belonging to the cbl B group, while activity increased severalfold in other cbl B mutants and in all cbl A, cbl C, and cbl D mutants examined, although mutase activity was still less than 10% of control. Comparison of mutase holoenzyme activity and total propionate pathway activity suggests that enhancement of mutase activity in mutant cells after cobalamin supplementation to values 5--10% of control may be sufficient to overcome the inherited metabolic block and to restore total pathway activity to normal.


Interaction of cations with phosphate uptake by Saccharomyces cerevisiae. Effects of surface potential. The effect of bivalent cations on phosphate uptake by Saccharomyces cerevisiae was investigated. Phosphate uptake via the Na+-dependent transport system at pH 7.2 is stimulated by bivalent cations. The apparent affinity of phosphate for the transport mechanism is increased, but the apparent affinity for Na+ is decreased. Uptake of phosphate via the Na+-independent transport system is accompanied by a net proton influx of 2H+ and an efflux of 1 K+ for each phosphate ion taken up. At pH 4.5 phosphate uptake via the Na+-independent system is stimulated by bivalent cations, whereas at pH 7.2 uptake is inhibited. The effect of bivalent cations on phosphate uptake can be ascribed to a decrease in the surface potential.


Characterization of prolactin binding by membrane preparations from rat liver. Binding sites for prolactin were identified in a plasma-membrane-enriched fraction isolated from livers of mature female rats. 125I-labelled sheep prolactin prepared by the lactoperoxidase procedure retained the same molecular integrity and binding affinity as the native hormone at physiological pH. The receptors bound prolactin from different species, whereas non-lactogenic hormones were not bound. The binding of 125I-labelled sheep prolactin was activated equally by bivalent and univalent cations, bivalent cations exerting their maximal effect at much lower concentrations. The association of 125I-labelled sheep prolactin with the receptor was a time- and temperature-dependent process. Partial dissociation was detected. The binding of 125I-labelled sheep prolactin was strongly influenced by pH, with an optimum observed at pH 6.5. Receptor activity was destroyed by Pronase and phospholipase C, whereas neuraminidase increased binding. Treatment of the membranes by ribonuclease and deoxyribonuclease did not affect the binding. Binding of 125I-labelled sheep prolactin was inhibited by p-chloromercuribenzoic acid, dithiothreitol and by brief exposure to high temperatures. Scatchard analysis of the binding of 125I-labelled sheep prolactin to receptors indicated that prolactin has a high affinity for its receptor. Binding of prolactin to liver membranes showed some properties different from those observed with mammary cells. Binding by these tissues differed in pH optimum, in effects of ions, and in response to neuraminidase.


Choline kinase and ethanolamine kinase activity in the cytosol of nerve endings from rat forebrain. Both choline kinase and ethanolamine kinase are present in the cytosol of nerve endings prepared from rat brain are the products of their action, phosphocholine (84 nmol/g fresh wt. of brain) and phosphoethanolamine (190 nmol/g fresh wt. of brain). In contrast with the enzymes from the cytosol of whole brain, both are as equally active at pH 7.5 as 9.0. Determination of kinase activity in membrane-containing tissue samples at pH9 gives low values because of the activity of alkaline phosphatase. Choline kinase, but not ethanolamine kinase, requires Mg2+ in excess of that required for the formation of the MgATP complex and is inhibited by an excess of free ATP. The Km for choline is 2.6mM and for ethanolamine is 2.2mM. The differing requirements for ATP and Mg2+ and the inhibition of choline kinase, but not ethanolamine kinase, by hemicholinium-3 suggest either the presence of two separate enzymes or two different active sites on the same enzyme.


Spectroscopic and kinetic properties of the transient intermediate acceptor in reaction centers of Rhodopseudomonas sphaeroides. The photoreductive trapping of the transient, intermediate acceptor, I-, in purified reaction centers of Rhodopseudomonas sphaeroides R-26 was investigated for different external conditions. The optical spectrum of I- was found to be similar to that reported for other systems by Shuvalov and Klimov ((1976) Biochim. Biophys. Acta 400, 587--599) and Tiede et al. (P.M. Tiede, R.C. Prince, G.H. Reed and P.L. Dutton (1976) FEBS Lett. 65, 301--304). The optical changes of I- showed characteristics of both bacteriopheophytin (e.g. bleaching at 762, 542 nm and red shift at 400 nm) and bacteriochlorophyll (bleaching at 802 and 590 nm). Two types of EPR signals of I- were observed: one was a narrow singlet at g = 2.0035, deltaH = 13.5 G, the other a doublet with a splitting of 60 G centered around g = 2.00, which was only seen after short illumination times in reaction centers reconstituted with menaquinone. The optical and EPR kinetics of I- on illumination in the presence of reduced cytochrome c and dithionite strongly support the following three-step scheme in which the doublet EPR signal is due to the unstable state DI-Q-Fe2+ and the singlet EPR signal is due to DI-Q2-Fe2+. : formula: (see text), where D is the primary donor (BChl)2+. The above model was supported by the following observations: (1) During the first illumination, sigmoidal kinetics of the formation of I- was observed. This is a direct consequence of the three-sequential reactions. (2) During the second and subsequent illuminations first-order  (exponential) kinetics were observed for the formation of I-. This is due to the dark decay, k4, to the state DIQ2-Fe2+ formed after the first illumination. (3) Removal of the quinone resulted in first-order kinetics. In this case, only the first step, k1, is operative. (4) The observation of the doublet signal in reaction centers containing menaquinone but not ubiquinone is explained by the longer lifetime of the doublet species I-(Q-Fe2%) in reaction centers containing menaquinone. The value of tau2 was determined from kinetic measurements to be 0.01 s for ubiquinone and 4 s for menaquinone (T = 20 degrees C). The temperature and pH dependence of the dark electron transfer reaction I-(Q-Fe2+) yields I(Q2-Fe2+) was studied in detail. The activation energy for this process was found to be 0.42 eV for reaction centers containing ubiquinone and 0.67 eV for reaction centers with menaquinone. The activation energy and the doublet splitting were used to calculate the rate of electron transfer from I- to MQ-Fe2+ using Hopfield's theory for thermally activated electron tunneling. The calculated rate agrees well with the experimentally determined rate which provides support for electron tunneling as the mechanism for electron transfer in this reaction. Using the EPR doublet splitting and the activation energy for electron transfer, the tunneling matrix element was calculated to be 10(-3) eV. From this value the distance between I- and MQ- was estimated to be 7.5--10 A.


Properties of ATP-driven reverse electron flow in chloroplasts. 1. The reverse reactions induced by coupled ATP hydrolysis were studied in spinach chloroplasts by measurements of the ATP-induced increase in chlorophyll fluorescence reflecting reverse electron flow, and of the ATP-induced decrease in 9-aminoacridine fluorescence, representing formation of the transthylakoidal proton gradient (deltapH). ATP-induced reverse electron flow was kinetically analysed into three phases, of which only the second and third one were paralleled by corresponding phases in deltapH formation. The rapid first phase and formation of a deltapH occur also in the absence of the electron transfer mediator phenazine methosulfate. 2. The rate and extent of the reverse reactions were measured at temperatures in the range from 0 to 30 degrees C. The rate of formation of delta pH and of reverse electron flow were faster at high temperatures, but the maximal extent of delta pH and chlorophyll fluorescence increase were observed at the lowest temperature. Considering rate and extent of the ATP-stimulated reactions, a temperature optimum around 15 degrees C was found. Light activation of the ATPase occurred throughout the range studied. At 0 degrees C and in the presence of inorganic phosphate the activated state for ATPase was maintained for more than 10 min. 3. The ATP-induced rise in chlorophyll fluorescence yield was found to be of similar magnitude as the rise induced by 3-(3,4-dichlorophenyl)-1,1-dimethyl-urea (DCMU), when both were measured with an extremely weak measuring beam. It is concluded, that both effects, although derived via distinctly different pathways, are limited by the same electron donating or electron accepting pool.


Quantitative estimation of the photosynthetic proton binding inside the thylakoids by correlating internal acidification to external alkalinisation and to oxygen evolution in chloroplasts. The external alkalinisation delta pHe, or the rate of oxygen evolution vO2, of a suspension of envelope-free chlorplasts was correlated with their internal acidification, estimated from the transmembrane delta pHei. Knowing the external buffer value, the concentration of the total protons moved Hi was calculated from the delta pHe, measured with a glass electrode ([Hi] was also obtained from vO2), and the free proton concentration [Hi+] was determined from delta pHei, measured with 9-aminoacridine. This gives a ratio gamma i = theta [Hi]/theta [Hi+], which is independent of the thylakoids internal volume. Within a large pHi range, scanned by varying the light intensity, gamma i was kept reasonably constant; it was hardly sensitive to pHi. This apparent invariability implies a continuous change of the internal buffer value beta i with pHi, since beta i/gamma i = -2.3.....10pHi, a relationship which inlcudes neither the total concentration of protonizable groups [Ai] nor pKi. As gamma i approximately Ki[Ai]/(Ki + [Hi+i]2, to keep gamma i constant when pHi drops, pKi and [Ai] must increase. This may be achieved by a progressive unmasking of anionic functions, initially inaccessible in the membrane. The relative slowness of this process may explain why gamma i calculated from the initial kinetics was sometimes smaller in high than in low light, where it always equalled that measured from the steady-state amplitude at all intensities. A small deficit of [Hi+] deduced from what could have been expected from delta pHe may reflect a limited binding of protons in the membrane itself, about 1 H+ for 30--130 chlorophylls (gamma i could be between 70 and 240, more frequently around 100); these numbers varied depending on the samples, but were constant for a given preparation.


The plastoquinone pool as possible hydrogen pump in photosynthesis. The function of the plastoquinone pool as a possible pump for vectorial hydrogen (H+ + e-) transport across the thylakoid membrane has been investigated in isolated spinach chloroplasts. Measurements of three different optical changes reflecting the redox reactions of the plastoquinone, the external H+ uptake and the internal H+ release led to the following conclusions: (1) A stoichiometric coupling of 1 : 1 : 1 between the external H+ uptake, the electron translocation through the plastoquinone pool and the internal H+ release (corrected for H+ release due to H2O oxidation) is valid (pHout = 8, excitation with repetitive flash groups). (2) The rate of electron release from the plastoquinone pool and the rate of proton release into the inner thylakoid space due to far-red illumination are identical over a range of a more than 10-fold variation. These results support the assumption that the protons taken up by the reduced plastoquinone pool are translocated together with the electrons through the pool from the outside to the inside of the membrane. Therefore, the plastoquinone pool might act as a pump for a vectorial hydrogen (H+ + e-) transport. The molecular mechanism is discussed. The differences between this hydrogen pump of chloroplasts and the proton pump of Halobacteria are outlined.


Potassium-stimulated ATPase activity and hydrogen transport in gastric microsomal vesicles. The Mg2+-dependent, K+-stimulated ATPase of microsomes from pig gastric mucosa has been studied in relation to observed active H+ transport into vesicular space. Uptake of fluorescent dyes (acridine orange and 9-aminoacridine) was used to monitor the generated pH gradient. Freeze-fracture electron microscopy showed that the vesicular gastric microsomes have an asymmetric distribution of intramembraneous particles (P-face was particulate; E-face was relatively smooth. Valinomycin stimulated both dye uptake and K+-ATPase (valinomycin-stimulated K+-ATPase); stimulation by valinomycin was due to increased K+ entry to some intravesicular activating site, which in turn depends upon the accompanying anion. Using the valinomycin-stimulated K+-ATPase and H+ accumulation as an index, the sequence for anion permeation was NO-3 greater than Br- greater than Cl- greater than I- greater than acetate approximately isethionate. When permeability to both K+ and H+ was increased (e.g using valinomycin plus a protonophore or nigericin), stimulation of K+-ATPase was much less dependent on the anion and the observed dissipation of the vesicular pH gradient was consistent with an 'uncoupling' of ATP hydrolysis from H+ accumulation. Thiocyanate interacts with valinomycin inhibiting the typical action of the K+ ionophore. But stimulation of ATPase activity was seen by adding 10 mM SCN- to membranes preincubated with valinomycin. From the relative activation of the valinomycin-stimulated K+-ATPase, it appears that SCN- is a very permeant anion which can be placed before NO-3 in the sequence of permeation. Valinomycin-stimulated ATPase and H+ uptake showed similar dependent correlations, including: dependence on [ATP] and [K+], pH optima, temperature activation, and selective inhibition by SH- or NH2-group reagents. These results are consistent with a pump-leak model for the gastric microsomal K+-ATPase which was simulated using Nernst-Planck conditions for passive pathways and simple kinetics for the pump. The pump is a K+/H+ exchange pump requiring K+ at an internal site. Rate of K+ entry would depend on permeability to K+ as well as the counterion, either (1) the anion to accompany K+ or (2) the H+ efflux path as an exchange ion. The former leads to net accumulation of H+ and anion, while the latter results in non-productive stimulation of ATP hydrolysis.


The influence of charge on phosphatidic acid bilayer membranes. A complete titration of phosphatidic acid bilayer membranes was possible for the first time by the introduction of a new anaologue, 1,2-dihexadecyl-sn-glycerol-3-phosphoric acid, which has the advantage of a high chemical stability at extreme pH values. The synthesis of the phosphatidic acid is described and the phase transition behaviour in aqueous dispersions is compared with that of three ester phosphatidic acids; 1,2-dimyristoyl-sn-glycerol-3-phosphoric acid, 1,3-dimyristoylglycerol-2-phosphoric acid and 1,2-dipalmitoyl-sn-glycerol-3-phosphoric acid. The phase transition temperatures (Tt) of aqueous phosphatidic acid dispersions at different degrees of dissociation were measured using fluorescence spectroscopy and 90 degrees light scattering. The Tt values are comparable to the melting points of the solid phosphatidic acids in the fully protonated states, but large differences exist for the charged states. The Tt vs. pH diagrams of the four phosphatidic acids are quite similar and of a characteristic shape. Increasing ionisation results in a maximum value for the transition temperatures at pH 3.5 (pK1). The regions between the first and the second pK of the phosphatidic acids are characterised by only small variations in the transition temperatures (extended plateau) in spite of the large changes occurring in the surface charge of the membranes. The slope of the plateau is very shallow with increasing ionisation. A further decrease in the H+ concentration results in an abrupt change of the transition temperature. The slope of the Tt vs. pH diagram beyond pK2 becomes very steep. This is the result of reduced hydrocarbon interaction energy, which was demonstrated by differential scanning calorimetry (Blume, A. and Eibl, H., unpublished data).


Active K+ transport in Mycoplasms mycoides var. Capri. Relationships between K+ distribution, electrical potential and ATPase activity. The addition of 5 . 10(-5) M or less of dicyclohexylcarbodiimide to Mycoplasma mycoides var. Capri preferentially influences K+ influx rather than efflux and reduces by 30--40% the activity of the membrane-bound Mg2+- ATPase. Adding valinomycin to metabolizing cells does not markedly affect K+ distribution but induces a rapid and complete loss of intracellular K+ in non-metabolizing cells. Uncoupling agents such as dinitrophenol, carbonylcyanide p-trifluoromethoxyphenylhydrazone, dissipate the K+ concentration gradient only when combined with valinomycin. Variations in the merocyanine fluorescence intensity indicate that a transmembrane electrical potential (delta psi) is generated on cell energization. This delta psi, not affected by valinomycin or uncouplers when used alone, is collapsed by a mixture of both. No change in fluorescence intensity can be detected when glucose is added to dicyclohexylcarbodiimide treated organisms. These experiments suggest that the membrane-bound Mg-ATPase activity control K+ distribution in these organisms through the generation of a transmembrane electrical potential difference.


Enzymatic reduction and methylation of folate following pH-dependent, carrier-mediated transport in rat jejunum. Intestinal transport of [3H] folate was studied using everted sacs of rat jejunum. The proximal small intestine transports folate against a concentration gradient by a system which is saturable, pH-dependent, energy-dependent, sodium-dependent, sensitive to temperature, and appears to be a common transport system for folate compounds. Chromatographic analysis of folate compounds in the serosal compartment after a 60 min incubation with folate in the mucosal medium in sodium phosohate buffer indicated that metabolism of folate to 5-methyltetrahydrofolate was extensive at pH 6.0 and negligible at pH 7.5. The percent conversion of folate to 5-methyltetrahydrofolate at pH 6.0 was reduced by increasing the concentration of folate in the mucosal medium, thus indicating saturation of the reduction and methylation process. These findings indicate that folate transport in rat jejunum occurs by an energy-dependent, carried-mediated system and that both folate transport and intestinal conversion of folate to 5-methyltetrahydrofolate are pH-dependent.


Phosphatidylseryl-transfer RNA in Bacillus licheniformis 749/C. Phosphatidylserine has been found in extracts of Bacillus licheniformis made under alkaline conditions but not under neutral or acidic ones and was derived from the tRNA fraction. In tRNA preparations kept below neutrality during purification, phosphatidylserine was the only phospholipid released when the pH was raised to 9.0. The amount of bound phosphatidylserine could be increased by incubating tRNA from B. licheniformis or Escherichia coli with CTP and phosphatidic acid in the presence of an S-30 extract from either organism. The tRNA carrying phosphatidylserine has been separated from the bulk of the tRNA by DEAE-Sephadex chromatography in the presence of a detergent. On deaminoacylation of this material and rechromatography on DEAE-Sephadex, a number of peaks were found, indicating that this behavior is not confined to a single isoaccepting species.


Polynucleotides XLVII. Synthesis and properties of poly(2-methylthio- and 2-ethylthioadenylic acid). Formation of non-Watson-Crick type complexes. Poly(2-methyl- and 2-ethylthioadenylic acid) were prepared by polymerization of corresponding diphosphates with Escherichia coli polynucleotide phosphorylase. These polynucleotides have relatively large hypochromicity of 30-35%. Acid titration of these polymers showed abrupt transition at pH 5.34-5.4, which may indicate that the introduction of alkylthio group at 2-position of adenine bases reduced their basicity. Thermal melting of these polymers showed no clear transition points at neutral pH, but in acidic media they have Tm values of 57 and 56 degrees C, somewhat lower than that of poly(A). Upon complex formation with poly(U), these poly(A) analogs showed only one poly(rs2A) . poly(U) type double-strand complexes, similar to that found in the case of poly(m2A) . poly(U).


Identification, partial purification and biochemical characterization of gamma-glutamyltranspeptidase present as a membrane component in skimmed milk and milk fat-globule membranes, and in mammary-tumour virus from the milk of infected mice. The enzyme gamma-glutamyltranspeptidase was reproducibly found to be associated with mouse milk particles; it is present in milk fat-globule membranes and mouse mammary-tumour virus of infected Swiss mice, also in particles from the milk of uninfected mice. The enzymatic activities observed range among the highest reported for mammalian tissues. The enzyme was partially purified from mouse mammary-tumour virus, and from milk fat-globule membranes. The molecule requires the presence of detergents to remain soluble, behaves as a high molecular weight component, properties characterizing integral membrane proteins. Kinetics, and the effect of competitors as well as of specific inhibitors show this enzyme to be identical to the well-known kidney gamma-glutamyltranspeptidase ((gamma-glutamyl)-peptide:amino-acid gamma-glutamyltransferase, EC 2.3.2.2). Other oncornaviruses budding from cultured cells originally expressing the enzyme in their plasma membrane also incorporate the enzyme in their structure.


Reversible inactivation of (Na+ + K+)-ATPase by use of a cleavable bifunctional reagent. 1. Purified (Na+ + K+)-ATPase, prepared from rabbit kidney outer medulla, is incubated with the bifunctional NH2-directed reagent dimethyl 3,3'-dithiobis-propionimidate. This results in a cross-link between the subunits of the enzyme and a simultaneous reduction of the (Na+ + K+)-ATPase and K+-stimulated p-nitrophenylphosphatase activities. 2. The most abundant cross-link product is a dimer of the two different subunits of the enzyme. 3. Reduction of the disulfide cross-link by dithioerythritol results in partial recovery of the original subunit structure of the enzyme and of the (Na+ + K+)-ATPase and K+-stimulated p-nitrophenylphosphatase activities. 4. These results suggest that a free mobility of the subunits of the (Na+ + K+)-ATPase system relative to each other is essential for proper functioning of both enzyme activities.


The inhibition of stearoyl-coenzyme A desaturase by phenyllactate and phenylpyruvate. The inhibition of oleic acid from stearic acid was studied using a rat liver microsomal preparation. Phenyllactic acid and phenylpyruvic acid are partially inhibitory. The may be related to the low oleic acid to stearic acid ratios found in brains of phenylketonuric persons and experimentally phenylketonuric rats.


Magnetic circular dichroism studies of bovine liver catalase. Absorption, circular dichroism (CD) and magnetic circular dichroism (MCD) spectra of beef liver catalase at pH 5.0 and 6.9, and its complexes with NaF, KCNO, NaCNS, NaN3 and NaCN, have been measured between 250 nm and 700 nm at room temperature. The pH 6.9 native catalase MCD shows the presence of several additional transitions not resolved in the absorption spectrum. While these bands can be seen in the spectra of all the derivatives, with the exception of the cyanide, their relative intensities changes considerably between complexes. Of special interest in the MCD of ferric hemes is the signal intensity at about 400 nm and 620 nm. The data indicate that the MCD intensity at 620 nm increases as the high spin iron porphyrin fraction increases, reaching a maximum with the fluoride complex. The 430 nm band intensity increases as the proportion of low spin iron increases, reaching a maximum with the cyanide complex. The MCD spectra also indicate clearly the existence of spin mixtures in the complexes with CNO-, CNS-, and N3-, where both the 430 nm and 620 nm bands have appreciable intensity. It is significant that despite almost identical absorption spectra the CNS- complex has higher fraction of low spin iron than either the CNO- or the N3- species. The differences between the pH 5 and 6.9 MCD spectra of the native catalase suggest that the environment of the heme centre is sensitive to protonation.


Purification and preliminary characterization of two asclepains from the latex of Asclepias syriaca L. (milkweed). Two groups of asclepains have been isolated from Asclepias syriaca L. (milk-weed) latex and a representative of each has been purified. Asclepains A3 and B5 are homogeneous proteins with molecular weights of 23 000 and 21 000, respectively. Both require a reducing and chelating agent for maximum activity and hydrolyze ester, amide and peptide bonds. The optimum pH for hydrolysis of casein is 7.5 to 8.5 for asclepain A3 and 7.0 to 7.5 for asclepain B5. Both enzymes are autolytic when active and are inhibited by p-chloromercuribenzoate, iodoacetic acid and sodium tetrathionate. Asclepains A3 and B5 each contain one titratable SH group per molecule and no bound carbohydrate. Each of the two enzymes has leucine as the N-terminal amino acid. There are notable differences in their amino acid compositions.


The destruction of serine and threonine thiohydantoins during the sequence determination of peptides by 4-N,N-dimethylaminoazobenzene 4'-isothiocyanate. 1. A mechanism for the destruction of serine and threonine thiohydantoins during protein sequence analysis by the Edman-type degradation is proposed. The mechanism begins with the dehydration of serine and threonine side chains (at the cyclization stage) which occurs mainly in anhydrous acid solution. The dehydrated derivatives finally polymerize by way of the reactive methylene group (enamine) to form polymers with various molecular weights. In aqueous acid solution, the dehydrated thiohydantoins of serine and threonine undergo hydration (according to the Markovnikov rule) and ring fission, which leads to the irreversible breakdown of thiohydantoin ring. The serine derivative shows a much greater tendency to undergo these side-reactions than the threonine derivative. 2. In the presence of oxygen, the alkaline hydrolysis of amino acid thiohydantoins goes through an oxidation-deamination reaction at the C-N bond of the thiohydantoin ring and leads to the formation of thiourea derivative and keto acids. This reaction mechanism accounts for the low recoveries of amino acid obtained from the alkaline hydrolysis of amino acid thiohydantoins.


NADP-specific isocitrate dehydrogenase of Escherichia coli. IV. Purification by chromatography on Affi-Gel Blue. Affinity chromatography on Affi-Gel Blue has been used to purify the NADP-specific isocitrate dehydrogenase (EC 1.1.1.42) from Escherichia coli. The protocol permits rapid purification of the enzyme in milligram quantities with a yield of 50% and is carried out almost entirely at room temperature. The preparation was judged to be homogeneous by non-denaturing electrophoresis at pH 7.5 and denaturing electrophoresis in the presence of sodium dodecyl sulfate. The subunit molecular weight of 53 000, determined by sodium dodecyl sulfate gel electrophoresis, is in reasonable agreement with the value of 46 900 estimated from the amino acid composition data.


Conformational studies of oxytocin analogues with different ring sizes. I. Circular dichroism. Circular dichroic spectra were measured for three analogues of deamino-oxytocin of different ring sizes where the disulfide group of oxytocin is replaced by the (CH2)n group. Their backbone rings are composed of different numbers of atoms, i.e., they are nineteen, twenty and twenty-one for [1,6-aminopimelic acid]oxytocin (n = 1), [1,6-aminosuberic acid]oxytocin (n = 2) and [1,6-aminoazelaic acid]oxytocin (n = 3), respectively. The pH dependence of the circular dichroism spectra indicates that the conformation of [1,6-aminoazelaic acid]oxytocin is different from those of others and the temperature dependency reveals that the conformation of [1,6-aminopimelic acid]oxytocin is most rigid. [1,6-Aminosuberic acid]oxytocin is biologically most active among three derivatives and their biological activities are related to the conformation and internal motions of the peptide hormone analogues.


The effect of different zooxanthellae on the growth of experimentally reinfected hosts. 1. A method is given enabling the differential effects of different strains of zooxanthellae on host growth to be assessed. This technique uses the increase in the number of tentacles as the measure of growth. 2. Aposymbiotic polyps of the anemone Aiptasia pulchella reinfected with strains of Symbiodinium microadriaticum isolated from the anemone Aiptasia pulchella and the scyphozoan Cassiopea xamachana grow as well as normal Aiptasia polyps. 3. Aposymbiotic Aiptasia polyps reinfected with zooxanthellae from the gastropod Melibe pilosa and the clam Tridacna maxima grew no better than polyps lacking zooxanthellae. 4. These results lead to the conclusion that strains of zooxanthellae differ in their ability to enhance growth of Aiptasia polyps under the experimental conditions and that these differences may have important ecological consequences.


[Some properties of the catalytic subunit of adenosine 3':5'-monophosphate-dependent protein kinase from pigeon breast muscle]. A method for the preparation of a homogenous catalytic subunit of adenosine 3':5'-monophosphate-dependent protein kinase from pigeon breast muscle was developed. The molecular weight of the enzyme as determined by electrophoresis in the presence of sodium dodecyl sulfate was found to be 42000. The pH optimum of the catalytic subunit was around 8.0. The active site of the catalytic subunit was studied using some derivatives of ATP, containing different reactive groups in the triphosphate chain of the molecule. It may be assumed that the pH optimum of the enzyme inactivation by adenosine 5'-chloromethylpyrophosphonate and the protective effect of ATP suggest covalent binding of the imidazole ring in the enzyme active site. The kinetic mechanism of the protein kinase reaction was studied using the initial rate experiments and reaction product inhibition. The results obtained were consistent with a random Bi-Bi kinetic mechanism.


[Stability and specificity of extracellular protein inhibitor for trypsin from Actinomyces janthinus 118]. Some properties of protein inhibitor for trypsin (TI) from Act. janthinus 118 were studied. It was shown that TI has an antitrypsin activity within a wide pH range with a maximum at about 9,5. At 4 degrees and 20 degrees C TI is stable for 24 hours within the pH range of 6,0--11,0. At 100 degrees C TI is more stable in the slightly acid region of pH than at neutral or alkaline conditions. Trypsin and chymotrypsin inactivate the inhibitor for 8 hours. TI inhibits trypsin, fibrinolysin, subtilisin, pronase and terrilytin, but have no effect on chymotrypsin, thrombin, papain and pepsin. The dissociation constants for the trypsin-inhibitor complex were found to be 1,7.10-8 M, 4,1.10-9 M and 2,4.10-10 M, with casein, p-nitroanilide benzoylarginine and tosylarginine methyl ester used as substrates, respectively. The corresponding dissociation rate constants for the subtilisin-inhibitor complex were equal to 1.10-9 M and 4.10-10 M with casein and carbobenzoxy-L-alanyl-L-alanyl-L-leucin p-nitroanilide used as substrates, respectively.


[Light-dependent oxygen consumption by isolated pea chloroplasts under NADP+ photoreduction]. The exchange of oxygen during NADP+ photoreduction by isolated pea chloroplasts was studied. It was found that NADP+ oxidation is accompanied by oxygen photoreduction preceeding at a high rate. A possibility for calculation of the ration between the pseudocyclic electron transport and the total electron transport based on oxygen exchange in the presence of NaN3 and catalase, was established. It was found that the pseudocyclic transport can make up to 30% or more of total electron transport.


Urease bound to chitin with glutaraldehyde. The enzyme urease (urea amidohydrolase, EC 3.5.1.5) prepared from Cajanus indicus, has been immobilized with glutaraldehyde-treated chitin as the solid support. The immobilized enzyme was characterized by determining the pH profiles and optimum temperature. Effect of glutaraldehyde concentration on the binding of enzyme to chitin was studied. The storage stability of the chitin-urease system was determined.


[Effect of presynaptic neurotoxins from the bee and cobra venoms on spontaneous mediator secretion from the motor nerve endings in mice]. Phospholipases A2 (PhLA) from the bee and cobra venoms induced the three-phasic changes in miniature end-plate potential (MEPP) frequency recorded in the mouse diaphragm muscle: an initial fall of transmitter release followed by a transient increase before the final complete blockade. The removal of Ca2+ from the perfusing solution (below 10(-9) M) prevented the presynaptic effect of both PhLA. If all PhLA molecules were washed out by the Ca-free solution, the subsequent exposure to a standard solution (2 mM Ca2+) action provoked an increase in MEPP frequency. The agents capable of increasing the axoplasmic Ca concentration (ions K+, hypertonic sucrose and uncoupler--TTFB) caused the usual increase of MEPP frequency in the muscles treated with PhLA. Apparently, the presynaptic biockade induced by PhLA cannot be due to the depletion of transmitter stores.


[Effect of the agents of general anesthesia on mice after microwave irradiation]. The general anesthetics chloroform, ftorotan, sodium thiopental and sodium hydroxybutyrate (500 mg/kg) decrease the survival of mice irradiated by microwaves. Ether, ftorotan, hexenal and sodium thiopental administered immediately after irradiation reduce the time of ensuing side position by 8-14%, its duration being decreased by 15-48% as compared to the control.


Effects of oxygen tension and pH on the ultrasonic absorption properties of sickle cells. The ultrasonic absorption of sickle cells was compared with the morphological changes in these cells are produced by varying oxygen tensions and pH. A decrease in oxygen tension caused increased sickling and an increase in the ultrasonic absorption of the blood. An increase in pH reduced the ultrasonic absorption and decreased sickling. These findings indicate that ultrasonic absorption can detect changes in the aggregation state of sickle cell hemoglobin in situ within the red cell and that the method may be used to monitor the degree of sickling under various environmental conditions.


[Therapy of alcoholics from the viewpoint of physicians and psychotherapists]. In the treatment of alcoholism, the medical profession takes an important position which until now has not gained the necessary appreciation. The prognosis of alcoholism is more favourable than generally assumed, especially if it is possible to early recognize patients with alcohol problems. Besides pharmaco-therapeutic possibilities it is important that doctors also consider psychotherapeutic methods, especially psychotherapeutical procedures according to ROGERS and TAUSCH, as well as various methods of behavioural modification.


Action of histamine receptor agonists and antagonists on the rat uterus. 1 Histamine and a series of compounds acting selectively on H1- and H2-receptors were tested on the isolated oestrous uterus of the rat. 2 Histamine had a dose-dependent inhibitory effect on the contractions elicited by acetylcholine. This action was unaffected by H1-blockers but was competitively inhibited by H2-blockers. The H1-selective agonist, 2-(2-aminoethyl)thiazole was ineffective at doses 100 times greater than those of histamine. Conversely, all the H2-agonists showed activity in the order of potency: N'-methylhistamine greater than histamine greater than N'-N'-dimethylhistamine greater than 5-methylhistamine greater than 5-methyl-N'-methylhistamine. Among the non-imidazole compounds, dimaprit had an activity identical to that of histamine, but all the dimaprit-like compounds showed negligible activity. 3 The data obtained suggest that in the rat uterus, (a) the activation of H2-receptors is responsible for the inhibitory effect of histamine and its analogues; (b) the integrity of the histamine molecule seems to be less crucial than that of the dimaprit molecule for the maintenance of the H2-activity, since changes in its structure modify but do not abolish the biological activity as they do in the case of dimaprit; (c) the order of activity of the various H2-receptor agonists is different from that observed in other tissues.


Comparison of rat brain membrane antigens by complement mediated release of neurotransmitters from brain fractions using antisera against S-100 and Thy-1. Synaptosomal, glial and neuronal fractions were prepared from rat brain and incubated to accumulate radioactively labelled neurotransmitters. Treatment of these fractions with antisera and complement showed that anti-(Thy-1) serum gave good release (50-75% of total uptake) of all neurotransmitters tested from synaptosomal and glial fractions. GABA and glutamate were released from neuronal perikarya, but not norepinephrine or serotonin. Anti-(S-100) serum gave no significant release of any neurotransmitter tested from any of the fractions, although all of them had previously been shown to contain this protein. These results are compatible with the membrane nature of Thy-1 and the mainly soluble nature of S-100 protein. They permit a selection for membrane antigens and neurotransmitters from different brain fractions. Antigenic differences between neuronal and glial plasma membranes were suggested by results with antiserum raised against bulk-isolated rat neuronal perikarya.


Regional distribution of monoamines in the cerebral cortex and subcortical structures of the rhesus monkey: concentrations and in vivo synthesis rates. Endogenous monoamine concentrations and turnover rates vary markedly in different regions of neocortex as well as in various subcortical structures of young adult rhesus monkeys. Monoamine levels and synthesis rates in amygdala, hippocampus, neostriatum, thalamus and brain stem are generally similar to comparable measures previously reported in a variety of species. However, extending and confirming the results of an earlier study, cortical monoamines exhibit topographically specific patterns of distribution. Thus, dopamine concentration is highest in the prefrontal and temporal neocortex; it decreases along the fronto-occipital axis and only trace amounts are detectable in the visual cortex. The distribution of norepinephrine is similar to that of dopamine except that the highest concentrations of norepinephrine are found in somatosensory cortex instead of prefrontal cortex. The pattern of distribution of serotonin is more uniform. However, the distribution of its metabolite, 5-hydroxyindoleacetic acid, is complementary to that of dopamine: the concentration is lowest in prefrontal cortex and highest in posterior regions of the telencephalon. Synthesis of catecholamines as measured by DOPA accumulation in monkeys treated with an aromatic amino acid decarboxylase inhibitor, NSD 1015, generally parallels the distribution of the catecholamines while indoleamine synthesis, as measured by 5-HTP accumulation, is similar to the distribution of 5-hydroxyindoleacetic acid. It may be significant that synthesis rates for the catecholamines are especially high in various areas of association cortex.


Lactate administration and fatty liver and kidney syndrome development in biotin-deficient chicks. Two experiments were carried out to determine whether administration of lactate to biotin-deficient chicks induced fatty liver and kidney syndrome (FLKS). 2. The results suggest that increased serum lactate concentrations are a consequence of the syndrome rather than a contributory factor in its incidence. 3. The increase in liver lipids of birds affected by FLKS was not associated with an increase in the specific activity of the hepatic lipogenic enzyme acetyl CoA carboxylase accept when birds developed FLKS spontaneously in experiment 2. 4. Some biotin-deficient chicks did not show physical symptoms of deficiency although mean liver biotin concentrations were low (0.31 microgram/g liver).


Hydroxyl content of solution-precipitated calcium phosphates. A method is described for determination of the titratable hydroxide ion in calcium phosphate precipitates. The procedure requires accurate analysis of the other titratable species in the crystal lattice but is unaffected by the presence of other lattice constituents or impurities. The method was applied to precipitates that had been previously analyzed by solution thermodynamic techniques, and the results were consistent with the earlier observations. The hydroxide content of the precipitates increased with crystal maturity and with increasing pH of the precipitation medium. The hydroxide content of the amorphous phase and the immediate postamorphous-crystalline transformation phase was shown to be nearly zero. After 3 to 4 days' maturation, the hydroxide content of precipitates prepared at pH values of 7, 8, and 9 was shown to increase to approximately 23, 40, 56% of that required for pure hydroxyapatite.


Energy transduction in the mitochondrionlike bacterium Paracoccus denitrificans during carbon- or sulphate-limited aerobic growth in continuous culture. Paracoccus denitrificans was grown in carbon-limited aerobic continuous culture (critical dilution rate (Dc) = 0.48 h-1). The molar growth yield for carbon (succinate or malate) was constant at about 60 over a broad dilution range (growth rate) from 0.10 to 0.48 h-1. Measurements of the stoichiometry of proton translocation associated with the oxidation of endogenous substrates yielded a ratio of protons ejected from the cell per atom of oxygen consumed(leads to H+:O) of 8.55 which decreased to 5.85 in the presence of piericidin A (PA), a specific inhibitor of NADH dehydrogenase (EC 1.6.99.3). With starved cells, the observed leads to H+:O associated with the oxidation of added succinate in the presence of PA was 5.61. These observed leads to H:O's represent an underestimation since no correction was made for proton backflow during the short interval of respiratory activity. Aerobic growth of Pc. denitrificans in the chemostat becomes sulphate limited at entering concentrations of sulphate less than 300 is microM. Neither the maximum specific growth rate (measured at Dc) nor the observed molar growth yield for succinate decreased under sulphate limitation. The NADH oxidase in electron transport particles prepared from sulphate-limited cells was completely inhibited by PA. The stoichiometry of proton translocation associated with malate oxidation was similarly unaffected by sulphate limitation. It is concluded that (a) the respiratory chain of aerobic, heterotrophically grown Pc. denitrificans possesses three sites of energy conservation, including site III, (b) the number of protons ejected during the transfer of one pair of reducing equivalents along a region of the electron transport chain equivalent to a single energy-coupling site is 3, and (c) that sulphate limitation does not lead to a loss of proton translocation associated with the cytochrome-independent region of the respiratory chain.


Purification of UDP-N-acetylenolpyruvoylglucosamine reductase from Escherichia coli by affinity chromatography, its subunit structure and the absence of flavin as the prosthetic group. The enzyme UDP-N-acetylenolpyruvoylglucosamine reductase (EC 1.1.1.158) was purified to homogeneity from Escherichia coli by affinity chromatography on a NADP-agarose column. The evidence suggests that the enzyme (molecular weight 35,000) is composed of two nonidentical subunits of molecular weight 21,500 and 13,500, respectively. The absorption spectrum of the purified enzyme shows no absorption band around 450 nm and thus does not support the previous suggestions that the enzyme is a flavoprotein. However, the A280: A260 ratio gives a value of 0.86 which suggests the presence of tightly bound nucleotide. A quantitative transfer of tritium from 1,4-[4-3H]NADPH to UDP-N-acetylenolpyruvoylglucosamine to form UDP-N-E13H]acetylmuramic acid was also observed, which clearly shows that the enzyme is not a flavoprotein.


The effects of nitrilotriacetic acid on solubilities of zinc, copper, manganese, and iron in the stomach of sheep. Four sheep, each prepared with a rumen fistula and reetrant cannula in the proximal duodenum, were used to study the effects of ruminal administration of nitrilotriacetic acid on solubilities of zinc, copper, manganese, and iron in rumen and duodenal digesta. The sheep received a pelleted diet and were dosed with 0, 300, 600 and 1200 microgram of nitrilotriacetic acid per gram of diet via the rumen fistula. Higher concentration of soluble zinc, manganese, and iron but not copper, were found in the rumen of the sheep when they were dosed with nitrilotriacetic acid. The concentrations increased with increasing dose of the acid. However, only the solubilty of iron was increased in the duodenal digesta. Concentrations of soluble zinc and manganese in the rumen increased, whereas copper decreased, during the first 2 h after feeding. The pattern was reversed thereafter. Changes in the concentrations of soluble during 6 h afther feeding were comparatively small. It is concluded that the solubilty of iron in the stomach of sheep is increased by ruminal administration of nitrilotriacetic acid.


Production of an extracellular ribonuclease by Pseudomonas maltophilia. As part of a screening program for pseudomonad enzymes having an industrial interest, we selected ribonuclease (RNase) producing strains. Of the 150 pseudomonads screened, 6 were found to produce an extracellular RNase activity when grown on solid medium. In broth culture, the RNase activity from these six species remained bound to the cells unless gelatin was added to the medium. Gelatin was essential for the release of RNase in the broth culture, but the pH of the medium, addition of potential inducers such as nucleic acids, or addition of cations did not affect this release. However, gelatin did not appear to induce the synthesis of the enzyme. Strain B-88, identified as Pseudomonas maltophilia, was selected for further study of the enzyme. The extracellular RNase isolated from B-88 broth cultures could be separated in two fractions on the basis of the molecular weight by the ultrafiltration technique. The low molecular weight fraction reacts optimally at temperatures between 55 and 60 degrees C and optimal pH values varying from 7.4 to 9.5. At neutral or alkaline pH, the enzyme was stable at temperatures below 37 degrees C but was inactivated at 55 degrees C. The RNase was inhibited by mercury and cobalt and stimulated by magnesium.


Prolactin-secreting adenoma as part of the multiple endocrine neoplasia--type I (MEN-I) syndrome. Two patients presented with the galactorrhea-amenorrhea syndrome. One patient had previously had parathyroid hyperplasia and the other an insulinoma. Preoperative evaluation of each patient revealed hyperprolactinemia and radiological evidence of an abnormal sella turcica. Pituitary adenomas were identified and removed at surgery. Immunostaining techniques confirmed the presence of prolactin-containing cells in both tumors. We propose that prolactin-secreting tumors be considered as part of the MEN-I syndrome, and that patients presenting with the galactorrhea-amenorrhea syndrome be screened and followed sequentially for evidence of other endocrine neoplasia.


Mediastinal carcinoid with an incomplete Sipple's syndrome. A 60-year-old woman with a mediastinal carcinoid of probable thymic origin, widely metastatic, had associated parathyroid hyperplasia, medullary carcinoma of thyroid with amyloid stroma and an adrenal neuroma. This case demonstrated features of an incomplete Sipple's syndrome (MEA II). The relationship of these diverse tumors is consistent with the concept of the APUD system and neurocristopathies.


Effects of dietary vitamin B6 on the in vitro inactivation of rat tyrosine aminotransferase in host liver and Morris hepatomas. Control rats or rats bearing Morris hepatoma 5123C (intact), 5123C (adrenalectomized), 7794A, 7800, 8999, 9121, or 9618A were fed a purified diet either deficient or adequate for vitamin B6. The concentration of pyridoxal phosphate in the plasma, host livers, and hepatomas was determined, as well as the in vitro rate of inactivation of induced tyrosine aminotransferase in homogenates of host livers and hepatomas. The results demonstrated the presence of a cysteine-independent inactivating system for tyrosine aminotransferase in hepatomas 5123C (adrenalectomized), 7800, 8999, and 9121. Only in hepatoma 9121 was there a dramatic influence of the dietary vitamin B6 on the rate of cysteine-independent inactivation. A cysteine-dependent inactivating system for the enzyme was present in all host livers and hepatomas. The rate of this in vitro inactivation for both host livers and hepatomas apparently was a function of the concentration of pyridoxal phosphate, but inactivation of tyrosine aminotransferase occurred at a significantly lower concentration of pyridoxal phosphate in the hepatomas than in the host livers.


Effect of smoking on benzo(a)pyrene metabolism by human placental microsomes. Placentas were collected at term from a series of 21 women. Thirteen were smokers, and eight were nonsmokers. Microsomes were prepared and used in the following studies of benzo(a)pyrene metabolism: aryl hydrocarbon, hydroxylase, epoxide hydrase, high-pressure liquid chromatographic analysis of benzo(a)pyrene metabolites, and DNA binding. DNA-binding adducts were further characterized by Sephadex LH-20 chromatography. Aryl hydrocarbon hydroxylase activity was much higher in smokers than in nonsmokers. Epoxide hydrase activity with styrene oxide as the substrate showed no difference between smokers and nonsmokers. High-pressure liquid chromatographic analysis showed much greater formation of dihydrodiols, quinones, and phenols by microsomes from smokers. The amount of benzo(a)pyrene-7,8-dihydrodiol was almost equal to the amount of phenols produced by the microsomes of the smokers. Sephadex LH-20 analysis of DNA binding resulted in only one major benzo(a)pyrene-DNA adduct when microsomes from smokers were used; this peak corresponds to benzo(a)pyrene 7,8-diol-9, 10-oxide bound to DNA nucleoside(s).


Increased specific binding of [3H]diazepam in rat brain following chronic diazepam administration. Male rats (175 g) were given 30 mg of diazepam in their food daily for 35 days. The animals became drowsy and ataxic from this high dose of drug. After the 35-day dosing, the rats were killed daily, and specific binding of [3H]diazepam and [3H]flunitrazepam was determined in synaptosomal preparations from these and corresponding control rats. On days 3, 4, 6, and 7 after the treatment period the specific binding and specific binding of [3H]diazepam was double that of the control binding and specific binding of [3H]flunitrazepam was 1.67 times that of control. The data indicate that very high doses of diazepam, given for long periods, cause increased specific binding of radiolabeled ligand to brain subfractions. The possible mechanisms and implications are discussed. When lower doses or shorter dosage regimens are used, increased binding is not observed.


The monitoring of lysosomal integrity by pH-stat and light scattering measurements. Light scattering measurements were used to monitor the integrity of isolated rat kidney lysosomes during prolonged incubation at 37 degrees C or following the addition of lysolecithin. The fall in extinction at 520 nm (E520) was shown to correlate very well with the fall in the particulate enzyme activity and the corresponding rise in the soluble enzyme activity. Measurements were also made of the release of H+ from the lysosomes into the suspending medium following treatment with lysolecithin. A good relationship was obtained between acidification of the medium and changes in the light scattering (E520) of the lysosomal suspension. The value of these techniques in following rapid changes in the integrity of lysosomes is discussed.


[Quantitative determination of the activity of acid peptidases of industrial origin]. Reagent ninhydrine-Cd++, reacts with free alpha and epsilon amino groups of proteins. Horse-heart apomyoglobin was subjected to exhaustive succinylation, rendering the product non reactive to ninhydrine. The succinylglobin was submitted to enzyme digestion at pH 2.0, 4.0, 4.7 and 6.0. The commercially available enzymes contain mainly pepsin-like and chymosin-like enzymes. The enzymatic digests of succinyl-globin contain new free alpha-amino groups reacting with ninhydrin. Enzymatic digestion was performed under various condition (ratio E/S, pH). The results were compared to those obtained with synthetic substrate: PRO-HIS-LEU-SER-PHE(NO2)-NLEU-ALA-LEU-OME. The price of the synthetic substrate used, was more than 100 times the cost of succinyl-globin, thus the use of this substrate is a valuable tool for the quantitative estimation of peptidase activity in commercially available (pepsin, chymosin-like) enzymes.


DEAE-cellulose chromatography of creatine kinase isoenzymes--effect of pH and serum. DEAE-cellulose chromatography (pH 7.0) of human heart extracts revealed the presence of three creatine kinase isoenzymes. The CK3 (skeletal muscle) isoenzyme was not retained on the column under these conditions. The CK2 (heart) and CK1 (brain) isoenzymes eluted at a conductivity of 5.5 +/- 0.6 m omega-1 and 11.4 +/- 1.2m omega-1, respectively. When DEAE-cellulose chromatography was performed at pH 8.0, CK2 eluted at a slightly higher conductivity, 6.5 m omega-1, whereas CK1 eluted as before 12.0 m omega-1. DEAE-cellulose chromatography of CK2 and CK1 isoenzymes in the presence of serum protein, and serum albumin had no significant effect on the elution of CK2 at pH 7.0 and 7.4, and on the elution of CK1 at pH 7.0 and 8.0 However, serum and serum albumin decreased the affinity of CK2 for DEAE-celluose at pH 8.0, and caused this isoenzyme to elute at a conductivity of 3.0-3.5 m omega-1. The decreased affinity of CK2 for DEAE-cellulose was not due to aggregation of CK2 with albumin or some other serum protein, but was related to the amount of albumin applied to the column.


Relations between reported alcohol consumption and certain biological variables in an "unselected" population. We looked for variables that could serve as indexes of alcohol consumption. We tested iron, gamma-glutamyltransferase, mean erythrocyte volume, mean erythrocyte hemoglobin concentration, urea, uric acid, and alanine aminotransferase, among others, in an unselected population of men, and by principal components analysis we singled out the correlation between the two enzymes and the two hematologic values. On the other hand, calculation of the coefficients of correlation between the total amount of alcohol consumed and the 10 variables studied, together with multiple regression analysis, showed that the three variables that correlated most significantly with alcohol consumption were gamma-glutamyltransferase, mean erythrocyte volume, and the use of tobacco. Critical evaluation of the results leads us to conclude that still more discriminative biological indices msut be sought and that the use of psychosocial data is also desirable.


Isoelectric focusing of serum proteins on modified thin cellulose-acetate membranes. We applied a method previously described for treating cellulsoe acetate gel strips with boron trifluoride in methanol, to make them suitable for isoelectric focusing of proteins, to dry cellulose acetate membranes. The results obtained are shown to compare very favorably. Both wide and narrow pH gradients may be used and coupled either to conventional protein staining or to specific identification by immunofixation. Focusing ordinarily takes 90--120 min, is technically easy, and is economical in the use of ampholytes.


A highly sensitive, simple determination of serum iron using chromazurol B. A highly sensitive, simple determination of serum iron and binding capacity is described. FeIII/FeII reacts with chromazurol B (CAB) and cetyltrimethyl ammonium bromide (CTMA), the resulting substance being a highly coloured ternary complex. Maximal absorbance of the complex occurs at pH 4.6--5.5 at 630 nm. Lambert Beer's law holds between 0 and 80 mumol Fe/l. Molar absorptivity is 1.68 X 10(5) 1 . mol-1 . cm-1 at 630 nm. Interference by other serum components is negligible even at high concentrations.


Subcellular localization of gamma-glutamyltransferase activity in guinea pig liver. Effect of phenobarbital on the enzyme activity levels. The localization of gamma-glutamyltransferase activity in guinea pig liver was studied after subcellular fractionation. The enzyme activity was essentially connected with plasma membranes whereas only low activity was found in the endoplasmic reticulum. A similar activity distribution was demonstrated for 5'-nucleotidase. Highest specific activity of gamma-glutamyltransferase was found in plasma membranes enriched in bile canaliculi. In this fraction the specific activity was 35 times greater than the specific activity of the total homogenate, a value similar to the relative specific activity of (Na+,K+)-ATPase. More than 90% of the total gamma-glutamyltransferase activity in guinea pig liver was connected with parenchymal cells and the enzyme seemed to have an outside orientation. Animals treated with phenobarbital showed moderate increased in gamma-glutamyltransferase activity in serum and liver, whereas high activities were found in most bile samples. No particular liver subfraction showed substantial accumulation of gamma-glutamyltransferase activity. The present findings do not support the suggested use of serum gamma-glutamyltransferase measurements as a direct index of "microsomal enzyme induction".


Selective effects of clofibrate on alkaline phosphatase isoenzymes in serum. Administration of clofibrate to both hyperlipidaemic patients and normolipidaemic subjects produced a significant decrease, averaging 22%, in serum alkaline phosphatase activity. Quantitative isoenzyme analysis showed that this change was entirely attributable to an average reduction of 39% in the activity of liver alkaline phosphatase, and that no significant change in the bone isoenzyme occurred. An accompanying fall in serum gamma-glutamyltransferase activity was seen in some subjects but this change was not statistically significant in the group as a whole.


Effects of equiblocking doses of nadolol and propranolol on left ventricular performance. Nadolol, a recently developed noncardioselective beta-adrenergic blocker, has the potential advantages of a longer oral half-life (t 1/2) than propranolol and, in animal studies, markedly fewer direct myocardial depressant effects. Neither the relative intravenous potency of nadolol and propranolol nor the comparative effects of the 2 drugs on left ventricular performance has been studied in man. We compared equiblocking intravenous doses of nadolol and propranolol in 10 subjects with ischemic wall-motion disorders. Nadolol was on the average 6.2 times as potent on a milligram-for-milligram basis. Both drugs decreased resting heart rate (p less than 0.02) and produced small rises in both mean pulmonary artery (p less than 0.03) and mean pulmonary artery wedge (p less than 0.03) pressures without significantly reducing the cardiac output. Both drugs also produced depression of the radionuclide ejection fraction (p less than 0.002). There were no significant differences between the effects of the 2 drugs on any of the aforementioned variables. Thus, the effects of nadolol on left ventricular performances are similar to those of propranolol. Because of its long oral t 1/2, nadolol may prove to be a clinically useful drug.


Successful replantation of a forefoot.

Successful replantation of a forefoot. This is a report of replantation of a completely severed forefoot which was divided through the distal tarsus by a samurai sword cut. Primary reconstruction of bone, vessels, nerve and skin was carried out 2 hours following amputation. Neither the long extensor nor the long flexor tendons were repaired. Three years following replantation the patient had an excellent result. His foot appeared normal and there was no evident atrophy of the soft tissue of the foot. No claw toe deformity developed. Protective sensation and temperature sense returned and the 2 point discrimination on plantar surface of the toe was 15 mm. The patient can walk and run with a normal gait.


Reduced beta-adrenoceptor sensitivity in the elderly. The effect of age on sensitivity to both isoproterenol and propranolol has been investigated in 27 male volunteers aged 21 to 73 yr. The dose of isoproterenol (given as a rapid intravenous injection) required to increase the resting heart rate by 25 bpm (I25) increased with age. The I25 was repeated during an intravenous infusion of propranolol and the dose ratio (I25 after propranolol divided by the control I25) determined. This was related to the concentration of free propranolol in plasma. It was found that the effectiveness of any given free concentration diminished progressively with age. These data are consistent with a diminished responsiveness of the beta-adrenoceptor to both agonist and antagonist drugs with advancing years.


Influence of prazosin on the development of antinuclear antibodies in hypertensive patients. A recent publication suggested that antinuclear antibodies (ANA) occur in up to one third of patients treated with the hypotensive drug prazosin (Minipress). We have examined a large group of hypertensive patients and found ANA in 9.7% of 145 patients taking prazosin and in 12.2% of hypertensive patients on any treatment regimen. Excluding patients taking methyldopa (a drug associated with a high ANA incidence), the ANA incidence fell to 6.9% in the prazosin-treated group and to 7.2% of all treated hypertensives. Of over 350 untreated hypertensive patients, 6.0% had ANA. Prazosin did not increase the ANA incidence in patients on or off various drug combinations including beta blockers and diuretics. Analysis of the data by age and sex confirmed our conclusion that prazosin therapy is not accompanied by an increase in ANA.


The effect of temperature of concurrently administered fluid on the onset of ipecac-induced emesis. The time of onset of ipecac-induced emesis is not significantly influenced by the temperature of concurrently administered fluid. The average time of emesis with syrup of ipecac administered with cold (10 degrees C) versus warm (40 degrees C) water was found to be 30:59 and 30:18 min, respectively. The difference in induction time is not statistically or clinically significant.


Postischemic brain oxygenation with barbiturate therapy in rats. We measured rat brain cortex PO2 (PtO2) with gold microelectrodes (tip diameter 5--10 micron) for up to 2 hours after 16 min of transient global brain ischemia with and without thiopental 90 mg/kg infused iv over 60 min beginning at 5 min postischemia. Seventeen rats were immobilized and mechanically ventilated on 1% halothane in oxygen with continuous monitoring of PtO2, ECG, end-expiratory CO2, rectal temperature, and arterial blood pressure. Global ischemia was induced by trimethaphan hypotension to an MAP of about 50 torr and a neck tourniquet inflated to 1500 torr. Postischemia, nine control rats (11 PtO2 measurements) were untreated and eight rats (8 PtO2 measurements) received thiopental 90 mg/kg. Preischemia, PtO2 values in both groups ranged from less than 5--70 torr with values of greatest frequency between 10 and 15 torr. Postischemia, PtO2 in control rats peaked at 45 +/- 8 (SEM) torr at 20 min. In thiopental treated rats, peak PtO2 was 24 +/- 6 torr at 10 min postischemia. Relative frequency histograms of PtO2 revealed that PtO2 in thiopental treated rats was lower (p less than 0.05) between 15 and 30 min postischemia. The magnitude of the decrease in PtO2 between 105 and 120 min postischemia appeared to correlate directly with the absolute preischemic value (i.e., the higher the preischemic PtO2, the greater the decrease in PtO2 postischemia). These results suggest that thiopental administered in large doses in early postischemia does not improve brain oxygenation secondary to a reduction in brain oxygen consumption. The relevance of the correlation between the magnitude of the fall in PtO2 postischemia and the magnitude of the preischemic value is discussed.


Multivariate symptom analysis related to response to lorazepam treatment. In an attempt to explain the marked inter-trial differences in the percentage of patients reported to benefit from the anxiolytic agent lorazepam, a statistical analysis was carried out on the scores from a psychiatric symptom questionnaire, completed before and after treatment with lorazepam, by 69 psychiatric out-patients. Most of the patients presented with a mixture of different syndromes, with moderate to severe symptoms, and had already failed to respond to other treatment. Forty of the patients showed a favourable response to lorazepam. Multivariate analysis of the strength of 6 symptom clusters versus therapeutic response in this diagnostically heterogeneous patient sample revealed that, in addition to anxiety, there was a high relationship also between phobic and psychosomatic symptom scores and response to lorazepam. The relationship between depression scores and response was only of borderline significance, and that for hysteria and obsessionality was poor. It is suggested that one explanation of inter-trial differences in the percentages of patients improved by lorazepam may be that they are due to differences in the diagnostic composition of the patient populations studied.


Effects of azatadine maleate on subjective appraisal and psychomotor functions relevant to driving performance. Studies were carried out in normal healthy male subjects to assess the effects on psychomotor functions and subjective ratings of performance after acute administration of azatadine maleate, a potent antihistamine with additional antiserotonin activity. In the first trial, 2 mg azatadine was compared with another new antihistamine Sch 12169 (2 mg) and placebo. In a second trial, higher doses of azatadine (4 mg and 8 mg) were compared with dexchlorpheniramine (4 mg) and placebo. Both trials were of a double-blind, randomized Latin square design and subjects were assessed using a battery of tests, after administration of each trial drug. The time and sequence of tests were standarized, with a 1-week interval between test sessions. The results showed that azatadine did not produce significant impairment of psychomotor function at either the standard 2 mg or the maximum recommended 4 mg per day dosage level. Permormance was only significantly impaired, compared with that after placebo, at the 8 mg dose level and was of a similar order to that observed after dexchlorpheniramine at the usual 4 mg dosage. It is suggested, therefore, that at the normal recommended dosage of 2 mg per day, azatadine is not likely to impair driving ability.


The pH of sclerosing agents: a determinant of pleural symphysis. The rate of success in producing pleural symphysis with intrapleural instillation of sclerosing agents has been variable. Differences in the designs of studies probably account for some of the variability, but the reasons for the remainder are not clear. Since a low pH of the pleural fluid is associated with pleural adhesions and loculations, the pH of the commonly used solutions of sclerosing agents was determined, both in their usual concentrations and when diluted with large quantities of exudative pleural fluid. The buffered solution of tetracycline hydrochloride had the most acidic pH (2.0) and showed little change when diluted by pleural fluid. A 0.5 percent solution of sodium hydroxide had the highest pH (13.0). The remainder of the sclerosing solutions showed a range of pH from 4.3 to 8.7. Experimental and clinical experience suggests that tetracycline consistently has the highest rate of success in producing pleural symphysis. It appears that when proper technique is employed, the pH of the solution of the sclerosing agent is an important determinant of the production of pleural symphysis.


Effects of acid aspiration on pulmonary alveolar epithelial membrane permeability. Employing a modification of the in vivo model of a liquid-filled canine lung, we measured the movement of substances of specific sizes (albumin, 69,000 daltons with a molecular radius of 35 A; and dextran with a molecular weight of 150,000 to 170,000 and an approximate molecular radius of 100 A) from the pulmonary capillary blood to the liquid-filled lung. A solution with a specific pH (1.5 to 4.5) was instilled into the left lung of the animals at a dosage of 3 to 5 ml/kg of body weight. For both albumin and dextran with a molecular weight of 150,000 to 170,000, the time for 50 percent equilibration between the specific substance in the blood and the same substance in the pulmonary liquid decreased significantly with instillation of pulmonary liquid with a pH of 1.5 and 2.5 but did not with a pH of 3.5 or above (P less than 0.05). In addition, since histamine has been implicated as a possible humoral mediator leading to increased permeability of alveolar membranes, the levels of histamine were measured in pulmonary liquids and blood in all groups. Levels of histamine in the pulmonary liquid (but not blood) were significantly higher in animals with instillation of liquids with a pH of 1.5 and 2.5 compared to all other groups.


[Reduction of myocardial reperfusion damage using hypocalcemic, hyperkalemic, alkaline blood during post-ischemic oxygen resaturation]. Postischemic myocardial depression is largely caused by a reperfusion injury which can be avoided almost completely by initial reoxygenation with hypocalcemic, hyperkalemic, alkalotic blood. Reperfusate modification resulted in postischemic myocardial performance comparable to that achieved in hearts receiving continuous coronary perfusion.


[Comparison of the value of various methods for diagnosis of gastroesophageal reflux]. At the University Surgical Clinic Köln-Lindenthal 5 different methods were employed for assessment of gastro-esophageal reflux between 1975 - 1976 in 283 patients with and without reflux disturbances. Clinically, a gastro-esophageal reflux was found in 72% of the cases. The measurement of intra-esophageal pH with reflux-provocation was the most sensitive, and at the same time the most specific (83, 3%). Significant differentiation betwen patients with and without reflux who had an axial hiatus-hernia could not be made, using x-ray examination and counting of the number of times swallowed in the acid-clearing test. The most suitable procedure in proving the presence of gastro-esophageal reflux was esophagoscopy, intra-esophageal pH-measurement and acid-clearing-and-pH determination, in all of which the results correlated with each other to a high degree.


Cyclic nucleotides and the regulation of canine gastric acid secretion. The response of the cyclic nucleotide system (cAMP, cGMP, adenylate cyclase, guanylate cyclase, and specific phosphodiesterases) to two gastric acid secretagogues, histamine and acetylcholine, and two secretory inhibitors, prostaglandin E2 and secretin, was studied in vivo and in vitro in canine gastric fundic mucosa. Histamine and acetylcholine in vivo failed to stimulate cAMP but significantly increased cGMP; in vitro they affected neither adenylate cyclase nor guanylate cyclase. Prostaglandin E2 and secretin, however, increased cAMP in vivo and significantly stimulated adenylate cyclase in vitro. Specific phosphodiesterases were unaffected by these compounds. The changes, while not specifically localized to the acid-producing cells, are consistent with the suggestion that the control of canine gastric acid secretion may be mediated by changes in mucosal cAMP and cGMP.


Physiological and morphological characteristics of progressive disruption of the canine gastric mucosal barrier. The investigation had two major goals: to define the progression of physiological changes associated with disruption of the gastric mucosal barrier to sodium and hydrogen and to identify the morphological correlates of the physiological alterations. Fluxes of ions and water were determined before and after treatment of oxyntic mucosa with graded concentrations of butyric acid using dogs with gastric pouches. Three phases of barrier disruption were characterized: I, acceleration of normal Na+/H+ exchange; II, neutralization of H+; III, exudation of interstitial fluid. Parallel studies assessed morphological damage associated with these phases. In Phase I, cellular bulging into the lumen and dilation of intercellular spaces were evident. Some cellular erosion and extreme intercellular dilation were prominent in Phase II. Phase III was represented by necrotic changes and desquamation. It is concluded that disruption of transport mechanisms occurs sequentially and is closely correlated with morphological signs of progressive damage.


Hormonal regulation of perinatal enzyme differentiation in the mammalian liver. The adaptation of newborn mannals to extrauterine life depends in large part on the maturation of biochemical and physiological functions during perinatal development. Hormones such as glucocorticoids, catecholamines and glucagon can stimulate enzyme induction during development; on the other hand, insulin has been shown to antagonize these stimulatory effects. Only the surface of the problem of hormonal regulation of enzyme differentiation during the perinatal period has been reached, especially as regards human development. Each enzyme presents unique problems of chemical regulation; the functional consequences of these factors are not exactly the same in each tissue and perhaps not in each species. The possibility of using inducing agents such as hormones, drugs and substrates to promote biochemical enzyme differentiation is a new and exciting aspect which needs to be explored further as a means of facilitating survival and ensuring optimal extrauterine development of the immaturely born human infant or the full-term infant with delayed-enzymic development. However, any intervention in the carefully programmed interplay of different hormones which regulate normal enzymic adaptation and development during the perinatal period should be undertaken only after careful consideration. The possibilities of long-term harm must be weighed against short-term benefits.