Transport in isolated plasma membranes.

Transport in isolated plasma membranes. Plasma membrane vesicles constitute a simpler experimental system for studying transport compared to cells or intact tissue. The principal advantages of the vesicle approach are the elimination of metabolism as a complicating factor and the ability to control the composition of the solutions on both sides of the membrane. The major disadvantage is vesicle heterogeneity. However, techniques are available to avoid the kinetic artifacts that are due to the heterogeneity. Results of transport studies using membrane vesicles have conclusively shown that D-glucose and amino acids are co-transported with Na+ and that transport against a concentration gradient is driven by an electrochemical Na+ gradient. As a result of coupling between Na+ and the nonelectrolytes, estimates of the kinetic parameters of transport, Km and Vmax, require that the load on the Na+ gradient be taken into account. This has rarely been done. Although electrolyte transport is a major function of the plasma membrane, knowledge of the mechanisms involved is limited. Future investigations employing specific ionophores should contribute much to our understanding of the mechanisms underlying ATP-independent ion transport. Examples of the application of membrane vesicles for studying transport-related aspects of diseases are discussed.


Action of prostaglandins, endoperoxides, and thromboxanes on the lamb ductus arteriosus. Prostaglandin (PG) E2, the PG endoperoxides PGG2 and PGH2, and enzymatically generated PGI2 and thromboxane A2 (TXA2) were tested in vitro on circular strips of ductus arteriosus from mature fetal lambs. Both PGE2 and the PG endoperoxides produced a dose-dependent relaxation of the ductus at low PO2 (7-11 torr), and their action was reduced or abolished at high PO2 (410-660 torr). PGE2, however, was more potent than the endoperoxides. The reaction mixture containing PGI2 relaxed the hypoxic ductus, but this response was not due to PGI2 but to two, more stable and as yet unidentified, compounds, one of which is most certainly PGE2. TXA2 was inactive on the vessel at low and high PO2. These results confirm that PGE2 is the most effective PG acting on the ductus and provide further support to the hypothesis that this PG is responsible for patency of the vessel during fetal life. PGE2 action, however, may be complemented by that of another endoperoxide derivative formed in the PGI2 synthetic reaction which remains to be identified.


Heart rate response to isocapnic hypoxia in conscious man. We studied the effect of acute progressive hypoxia on heart rate in conscious healthy subjects. The PCO2 was held constant (+/- 1.5 mmHg) midway between the resting end-tidal and mixed-venous levels. Hypoxia was induced by having the subject rebreathe from a small bag so that the PO2 fell at a rate related to the subject's oxygen consumption. Arterial oxygen saturation (Sao2) was measured continuously during the procedure with an ear oximeter. We found that heart rate (HR) was best fitted to an inverse linear relation to arterial oxygen saturation and a power relation to PO2. The range of deltaHR/deltaSao2 was 0.62-1.46 beats/min per 1% fall in Sao2 (mean +/- SE = 0.98 +/- 0.06). There was no relationship between heart rate and ventilatory responses to hypoxia.


Effect of dihomo-gamma-linolenic acid on the canine pulmonary vascular bed. The effect of dihomo-gamma-linolenic acid (DGLA), the precursor of the monoenoic prostaglandins (PG), F1alpha and E1, on the pulmonary vascular bed of the intact dog was studied under conditions of controlled pulmonary blood flow. DGLA increased pulmonary vascular resistance in a dose-related manner by constricting intrapulmonary veins and upstream segments, presumably pulmonary arteries. Intrapulmonary injection of DGLA also increased transpulmonary injection of DGLA also increased transpulmonary airway pressure, presumably by increasing airway resistance and decreasing lung compliance or both. The vasoconstrictor response, however, was independent of changes in transpulmonary pressure since similar pressor responses were obtained in ventilated and nonventilated lungs. Further, the response was not dependent on factors or elements in whole blood, since the increase in pulmonary vascular resistance occurred during perfusion with saline or dextran and was enhanced in these media. Conversion of DGLA to PGs by a lung cyclo-oxygenase appears to mediate the response, since it was blocked by indomethacin and dose not occur with injection of nonprecursor long-chain fatty acids. These data suggest that the response to DGLA is due to formation of vasoactive products in the monoenoic PG pathway.


Relation of mitral valve opening and closure to left atrial and ventricular pressures in the intact dog. The exact times of mitral valve opening and closure were determined in dogs under varying hemodynamic conditions in 143 cardiac cycles (five experiments). Radiopaque markers had been sutured to the cusps and the valve annulus 7-124 wk before the studies. Valve opening and closure times were correlated with simultaneously obtained high-fidelity intracardiac pressures. Closure of the mitral valve was completed 5-105 ms after the atrial-ventricular pressure crossover; the time interval between the onset of ventricular systole and the instance of complete valve closure varied less (10-40 ms). These observations suggest that in the intact heart alpha, rapid mitral cusp closure at the end of diastole is initiated and completed by ventricular systole alone, and beta, the ventricular isovolumic contraction period might be shorter than assumed. Opening of the valve during ventricular relaxation was characterized by 1) initial separation of the markers placed on the free edges of the cusps, of variable duration, apparently due to alterations in ventricular geometry, and 2) a rapid opening motion which clearly preceded the diastolic pressure crossover by 5-60 ms. This finding would suggest that ventricular isovolumic relaxation might be shorter than generally accepted, although the mechanism of early opening is not adequately explained by the data.


Role of the descending pressor pathway in the conscious and pentobarbital-anesthetized dog. Resting cardiovascular parameters and the responses to bilateral carotid occlusions (BCO) were monitored in pentobarbital-anesthetized and conscious dogs before and after placing lesions in the dorsolateral funiculi at C7-C8 and after spinal transections at C7. Pre- and postlesion blood pressure (BP) and heart rate (HR) responses to exercise were also monitored. The lesions significantly attenuated the responses to BCO and decreased resting BP in anesthetized dogs. Yet neither resting HR in anesthetized or conscious dogs nor the resting BP in conscious dogs was affected by the lesions. Subsequent spinal transections significantly decreased resting HR and BP and the responses to BCO but did not affect the BP response to BCO in anesthetized dogs as compared with corresponding postlesion parameters. BP responses to exercise were significantly attenuated by the lesions, but HR responses were not affected. Since stimulation and BP studies indicated that the descending pressor pathway had been ablated, the data suggest that the pathway mediates BP and HR responses to BCO in pentobarbital-anesthetized and conscious dogs. It does not maintain resting HR in anesthetized or conscious animals, and the resting BP in conscious dogs. This pathway is important for BP responses to exercise but is not necessary for HR responses. Finally, other spinal pathways are involved in cardiovascular control.


Effect of regional myocardial ischemia on cardiac pump performance during exercise. The effect of brief periods of regional ischemia upon left ventricular pump performance was studied in nine dogs standing quietly at rest and during running exercise on a treadmill. Transient occlusions of the left circumflex coronary artery resulted in increase in heart rate at rest (+30 beats/min) but not during exercise. Other changes due to occlusion were similar at rest and during exercise and included decreases in stroke volume (-25% standing, -23% running); in dP/dt max, the maximum first derivative of the left ventricular pressure (-20% standing or running); and in left ventricular peak systolic pressure (-13% standing, -21% running); and rises in left ventricular end-diastolic pressure (+4.5 mmHg standing, +6.3 mmHg running). Cardiac output was unchanged by occlusions at rest but fell (-18%) during occlusions while the dogs were running. Propranolol reduced absolute levels of cardiac performance during exercise occlusions but had no effect at rest. Inotropic agents with ischemia had some effects at rest but did not alter exercise hemodynamics. It is concluded that integrated left ventricular function during ischemia is not impaired by exercise, probably because of beta-adrenergic stimulation of nonischemic myocardium.


Quantitation of anterior descending vs. circumflex venous drainage in the canine great cardiac vein and coronary sinus. The fractions of left anterior descending (LAD) and circumflex (LC) inflow drainage into the canine great cardiac vein (GCV) and coronary sinus (CS) have been quantitated by use of a right heart bypass preparation in which GCV outflow was isolated from the remainder of CS outflow. Following direct LAD injection of indocyanine green dye (ICG), 63 +/- 8% (SD) of the total amount of dye recovered appeared in GCV outflow and the remainder in CS outflow. CS recovery of ICG was decreased appreciably by ligation of epicardial venous connections between the LAD and LC beds, but was not affected by selective reductions of LAD or LC inflow. Only 3 +/- 3% of ICG injected into the LC was recovered in GVC outflow under basal conditions, and these low values were not affected measurably by selective reductions of LAD or LC inflow. CS drainage of LAD inflow could be augmented by selective increments of GCV pressure exceeding 7-10 mmHg. Increments of LC drainage in GCV outflow required CS pressures that exceeded GCV pressures by greater than 10 mmHg.


Direct effect of physiological doses of arginine vasopressin on the arterial wall in vivo. Effects of topically applied arginine vasopressin (VPA), in 100-150 muU/ml blood concentration, on external diameter (D) and on dynamic elastic modulus (E*) of surgically denervated common carotid (CC) and femoral (FA) arteries have been studied before and after hypophysectomy in anesthetized dogs. Changes in D, E*, and flow resistance (R) of the CC and FA vascular beds before and after removal of the pituitary were also determined. It was found that VPA elicited a substantial (max 21-26%) decrease in E* and a smaller (max 5.9-6.9%) reduction in D of FA independent of the presence of absence of the pituitary gland. The VPA effect developed more rapidly after hypophysectomy than before. In the CC, VPA did not significantly affect values of E*. During the 1-h period after hypophysectomy, statistically significant decreases in E*-CC, E*-FA, and D-CC were observed, but D-FA did not change, although arterial pressure as well as R-FA and R-CC were diminished. These results give further support to physiological vascular actions of VPA and a possible role in short-term circulatory control. In large arteries, the effects of VPA also seem to be regionally differentiated.


Antagonism of histamine edema formation by catecholamines. Histamine (4 microgram base/min) infused into the brachial artery for 60 min greatly increases skin lymph flow and lymph protein concentration in forelimbs perfused either naturally or at constant inflow. In contrast, the simultaneous intrabrachial infusion of histamine and norepinephrine (4 microgram base/min of each) or histamine and isoproterenol (4 microgram base/min and 3 microgram/min, respectively) for 60 min failed to alter lymph protein concentration in forelimbs perfused either naturally or at constant inflow. The edema in forelimbs naturally perfused by histamine (4 microgram base/min) infused into the brachial artery for 60 min was also greatly reduced by the simultaneous histamine-catecholamine infusions. Phentolamine (in concentrations that antagonize the vascular actions of norepinephrine) failed to prevent the antagonism of histamine protein efflux by norepinephrine. Hence, it must be concluded that catecholamines antagonize the protein efflux by locally infused histamine, independent of alpha-adrenergic receptor activity.


Reflux mechanisms involved in cardiac arrhythmias induced by hypothalamic stimulation. Electrical stimulation of widespread areas in the CNS has been shown to cause cardiac arrhythmias, which occur most frequently after cessation of stimulation. To determine the reflex and autonomic mechanism responsible for the poststimulation arrhythmias, we anesthetized cats with chloralose, and recorded arterial pressure, ECG, and cardiac vagal nerve activity. Stimulation of the hypothalamus consistently caused increases in blood pressure and heart rate during stimulation and caused arrhythmias, accompanied by vagal hyperactivity, immediately following stimulation. The arrhythmias were mediated solely by the vagus nerves because vagotomy or propantheline administration prevented them, whereas propranolol did not. Administration of either phentolamine or spinal cord transection prevented both the rise in blood pressure during stimulation and the poststimulation arrhythmias, but sectioning the carotid sinus and aortic depressor nerves had no preventative effect. However, when this denervation was combined with sectioning of vagal afferents, bursts of vagal activity (used as an index of cardiac rhythm disturbances) were prevented in three of six animals. Subsequent administration of phentolamine prevented the bursts in the remaining animals. It is concluded that poststimulation arrhythmias are elicited by the rise in blood pressure occurring during stimulation causing a sudden surge in parasympathetic outflow to the heart. The reflexogenic areas involved appear to be stretch receptors innervated by afferent vagal fibers.


Continuous measurement of renal blood flow changes to renal nerve stimulation and intra-arterial drug administration in the rat. A method is described for continuous measurement of renal blood flow in the anesthetized rat without dissection of the renal artery. Blood flow and arterial pressure were measured in an extracorporeal flow circuit between the carotid artery and an aortic pouch from which the left renal artery was the only outlet. Injection into the flow circuit allowed delivery of drugs directly into the arterial blood supply of the kidney. Electrical stimulation of undamaged periarterial renal kidney. Electrical stimulation of undamaged periarterial renal nerves was possible since the renal artery remained undisturbed. Extracorporeal autoperfusion of the rat kidney produced renal flow and resistance measurements that did not differ from those obtained with a flow probe placed directly on the renal artery. Renal nerve stimulation was found to cause renal vasoconstriction due to activation of alpha-adrenergic receptors by norepinephrine released from postganglionic sympathetic neurons. Renal vascular responses to a variety of intra-arterial vasoactive agents were also determined. The method described here allows the evaluation of renal vascular control in the variety of disease states for which suitable rat models have been developed.


Ethics in the practice of psychiatry--origins, functions, models, and enforcement. The author points out that psychiatric ethics are coming under increasing critical scrutiny by psychiatrists and by those outside of the profession. On the basis of an examination of APA Ethics Committee records, he concludes that criticism of psychiatry's inability to police itself might have some merit but cautions that the critics might be expecting too much from a code of ethics, which has the job of sensitizing psychiatrists to an ethical way of professional life and helping sort out ethical conflicts. After outlining a number of the problems in the self-policing aspect of ethics, the author makes suggestions for their resolution.


The psychiatrist's responsibility and the public trust. The author notes that psychiatrists alone can change the public image of their profession. This image has been tarnished in the past by grandiose claims of psychiatry's ability to right social wrongs, by the perceived lack of scientific underpinnings of the discipline, and by internal disputes within the profession. If they are to gain the public trust, psychiatrists must determine for themselves what unique skills they have to offer and how they can best provide their knowledge and expertise to the public in an effective and credible way.


Organic impairment in polydrug users: risk factors. In a national collaborative study to assess the neuropsychological status of 151 polydrug users, the Halstead-Reitan Neuropsychological Battery showed deficits in 37% two to three weeks after they entered treatment and in 34% at three-month follow-up. Comparative rates for a group of psychiatric patients were 26% and 27%, and for nonpatients, 8% and 4%. Extensive and intensive use of CNS depressants and opiates correlated positively with neuropsychological deficit. Older, less-educated subjects with adverse medical or developmental histories were more likely to show polydrug-related organic impairment. Although there is some evidence that such impairment is reversible, the condition appears to be of at least intermediate duration and may be long lasting.


Psychiatric consultation in a Vietnamese refugee camp. The authors provided psychiatric consultation to medical personnel in charge of a Vietnamese refugee camp in California. Although the emergency measures that supplied clothing, food, shelter, and medical care to nearly 60,000 refugees were immensely successful, consultation was requested to assist with selected mental health problems. The recommendations of the consulting team were related to the stage of camp development: early recommendations concerned easing adaptation to the camp setting; later efforts included creating a psychiatric crisis clinic and carrying out a mental health survey on a random sample of refugees. The results of this survey helped camp directors to understand how refugees responded to the camp experience.


Nocturnal penile tumescence and diagnosis in diabetic impotence. The authors conducted a study of nocturnal penile tumescence (NPT) in 35 diabetic men, aged 33 to 70, who complained of impotence and in 35 age-matched control subjects. EEGs and other measurements showed that the diabetic men as a group exhibited significant reductions in the total amount of NPT and in the amount and frequency of full erection, thus suggesting that impotence in this cohort was organogenic. Although NPT monitoring represents an advance over the less precise traditional procedures for the differential diagnosis of impotence, the authors stress the need for more research in this area.


Prediction research and the emergency commitment of dangerous mentally ill persons: a reconsideration. The author suggests that research on the prediction of violent behavior does not support the unqualified conclusion that the accurate predictions of violence is impossible under all circumstances or that psychiatrists, psychologists, and others will invariably overpredict its occurrence by several orders of magnitude. Further, he suggests that there are theoretical reasons why one could expect that one set of circumstances--those which typically apply in the short-term emergency commitment of mentally ill persons predicted to be imminently violent--may be exempt from the systematic inaccuracy found in the current research.


Emergency psychiatry and the assaultive patient. The authors studied 62 emergency room patients with violent ideation or action to assess the feasibility of predicting assaultive behavior. Although the likelihood of future violent acts could not be predicted accurately, they discovered that a significant majority of violent patients were potentially treatable within the setting of emergency psychiatry practice.


Psychiatrists' familiarity with legal statutes governing emergency involuntary hospitalization. The authors sent out questionnaires regarding knowledge of statutes governing emergency involuntary hospitalization to psychiatrists in Connecticut and in the District of Columbia. Fifty-five percent (N=207) of the sample in Connecticut and 25% (N=87) of the sample in the District of Columbia responded. The responses showed that few of these psychiatrists had a thorough familiarity with the legal statutes governing emergency involuntary hospitalization in their jurisdictions. The most frequently cited criterion was the dangerousness standard, especially in the District of Columbia sample, and many psychiatrists were not sure of the nature of their statutory authority when their patients met the criteria for emergency hospitalization.


Referrals from a psychiatric emergency room: relationship of compliance to demographic and interview variables. In a sample of 89 patients referred from a psychiatric service emergency room, the author investigated variables related to completion and noncompletion of the physician's recommendation. The completers (58%) tended to be older and more educated, to be diagnosed as depressed, and to demonstrate congruence in terms of their request and their perception of the physician's goal. Implications for referral include ensuring that the interview with the patient includes a communication, empathy, and mutual influence. Using the negotiated approach and criteria derived during the clinical interview, certain patients can be identified as being "at risk" for noncompletion and as requiring special efforts.


The assessment of depression: a model for quality review of emergency psychiatry. Of 167 patients appearing in the psychiatric emergency room of a metropolitan hospital, 68 were categorized as significantly depressed by the Zung depression scale but only 43 were considered depressed by clinician-interviewers. The use of a patient self-rating scale, which in this setting permitted the identification of deficiencies in the recognition of depression, is applicable to quality review in a variety of clinical settings.


High academic achievement in psychotic students. The authors studied 21 schizophrenic and borderline college students who achieved B+ or higher grade averages and underwent psychotherapy while in college. High academic achievement was found to provide relief from feelings of worthlessness and ineffectuality resulting from poor relationships with parents, siblings, and peers. Psychotherapy and the permissive yet supportive college atmosphere reinforced the students' self-esteem.


Effects of smoking on free recall and organization. Habitual smokers smoked either nicotine-free cigarettes or cigarettes containing a known amount of nicotine and then engaged in a free-recall task. Nicotine subjects recalled significantly fewer words on a 75-item list during three successive trials of immediate recall than did nicotine-free subjects. Contrary to expectations, the superiority of the nonnicotine group persisted over two days. The two groups displayed comparable organizational activity (indexed by category clustering).


Eye blink and schizophrenia: psychosis or tardive dyskinesia? The author found abnormalities of ocular movement including abnormal blink rate and blink reflex to glabellar tap in 34 of 44 medication-free schizophrenic patients. It is not clear whether these signs represent part of the pathology of schizophrenia or result from withdrawal of neuroleptic treatment.


A rating scale for emotional blunting.

A rating scale for emotional blunting. Although emotional blunting has always been considered a core symptom of schizophrenia, it has been excluded from recently developed sets of diagnostic criteria because of its alleged unreliability. The authors describe a brief rating scale for emotional blunting that is highly reliable, predicts short-term treatment response, and discriminates between patients with affective disorder and schizophrenia. They suggest that this scale will permit restoration of the important criterion of emotional blunting to modern diagnostic systems, including that proposed for DSM-III.


Splitting and identification with the aggressor in assaultive borderline patients. The author describes two borderline inpatients with assaultive behavior that was not controllable by medication. These patients use splitting to separate all-good and all-bad part representations, which hinders tolerance of ambivalence and control of aggression. Drive breakthroughs may reinforce their feelings of omnipotence. Interpreting episodes to the patient in this psychodynamic framework can aid their integration of self-representations and their ability to control aggression. Therapists and staff should resist the temptation to "give in" to such patients in order to forestall explosions.


Male urethritis in King County, Washington, 1974-75: I. Incidence. In a population-based study in King County, Washington an attempt was made to enumerate all cases of male urethritis receiving primary care from all sources over a defined period of time. The incidence of gonococcal (GCU) and non-gonococcal urethritis (NGU) was estimated to be 1,143 and 2,541 cases per year per 100,000 men age 15 and over, respectively, during 1974-1975. Private practitioners treat 62 per cent of GCU and 64 per cent of NGU estimated to occur in the county. They report only 3 per cent GCU which they see to the Health Department. Only 60 per cent of the general practitioners, 33 per cent of the internists and 65 per cent of the urologists had treated males with GCU in the three months prior to being questioned but these three groups of specialists see 95 per cent of all male urethritis treated by private practioners. The proportion of all male urethritis cases which were GCU ranged from .16 in a university student health clinic to .35 in private practices and Health Department clinics.


Male urethritis in King County, Washington, 1974-75: II. Diagnosis and treatment. Methods of diagnosing and treating male urethritis in King County. Washington in 1974-1975 were examined by a retrospective postal survey, records review, and prospective surveillance. A Gram-stained smear of urethral discharge was used by 86 per cent of private physicians and urethral culture by 56 per cent. Only 5 per cent made a diagnosis without laboratory aid. U.S. Public thealth Service gonorrhea treatment regimens were used by 54-81 per cent of private physicians. Over 90 per cent of physicians not in private practice (emergency room and housestaff) used smear, cultures, and recommended treatment regimens. The Gram-stained smear appeared to be under-utilized and the urethral culture overutilized. There is a significant need for physician education regarding recommended treatment for male gonorrhea.


Evaluation of drug dependence treatment in VA hospitals. A nationwide evaluation of 49 Veterans Administration drug dependence treatment programs was conducted using a sample of 1,655 patients. Their status with reference to seven treatment goals was assessed at admission and again 11 months later to determine change. The following changes were observed: a large decrease in heroin use; a small increase in the use of alcohol; moderate decline of involvement in the drug culture; moderate increase in psychological well-being; and small increase in economic independence. No statistically significant changes were observed in frequency of arrests, in establishment of stable living arrangements, in drug related medical problems, or in the ability to relate to other people. Implications of the findings for further research and program modification are discussed.


A factor analytic approach to an effective lung function screening protocol. Interrelationships among simple indices of pulmonary function derived from flow-volume curves and single-breath nitrogen tests were investigated by performing a factor analysis on data obtained from a generally healthy office working population (n = 388). Smoking was regarded as the major contribution to pulmonary dysfunction. Delta nitrogen was largely independent of all other measures, while flow-volume measures, including flow rates at high and low lung volumes, were highly intercorrelated but largely independent of closing volume and closing capacity. Delta nitrogen was most affected by smoking in both sexes. Scores derived from flow-volume measurements and from closing-volume measurements were significantly different, on the average, between male smokers and male nonsmokers. The methods and results presented prove to be useful in the design of protocols for effective pulmonary testing of large populations.


Community medicine in the British National Health Service. Reorganization of the British National Health Service in April 1974 brought about the creation of Community Medicine as a service specialty. Development of thinking on the type of role British community medicine should play is briefly described. The functions of the community medicine specialist at the different tiers of the health service are described in the context of the objectives of reorganization and the problems facing the specialty today are discussed.


Evaluation of the localization auditory screening test in children 6-18 months of age. The present paper is a report of a project to develop an automated auditory screening test for infants six to 18 months of age. The first year of the project was devoted to developing equipment and test procedures; the second year was concerned with testing the effectiveness of the equipment and procedures on an actual population of six to 18 month old infants. Two-hundred and fifty infants were screened auditorily as part of a county health department child development clinic. The pass/fail results of the screening test were evaluated in terms of physical and developmental examination following the screening and by means of a case review of the child's previous history. The results indicate that the procedure under investigation can be used to differentiate the normal hearing infant from the infant with possible hearing problems. It is shown by the test environment in which this study was conducted that the procedure reported can be successfully incorporated into a public health program, i.e., child development clinics or EPSDT programs.


Validity of 24-hour dietary recalls by adolescent females. The validity of 24-hour dietary recalls was tested by comparing the recalled intake of adolescent girls to their actual intake during a metabolic study. Diet recalls were a valid bases of estimating the dietary intake of energy, protein, calcium, and zinc by this group of adolescents. However, the majority of the subjects were unable to recall their food intake with enough accuracy so that their intake of vitamins A and C, thiamin, riboflavin, niacin, and iron could be calculated within the range of two-thirds to four-thirds of their actual intake.


The relative effects of lesser curvature vagotomy and esophageal vagotomy on the acid secretory effect of proximal gastric vagotomy. Proximal gastric vagotomy is an operation consisting of division of all vagal fibers to the acid-secreting portion of the stomach. These fibers are usually divided along the lesser curvature of the stomach; however, because of a high rate of duodenal ulcer recurrence in some series, it has become apparent that it is important to divide the vagal fibers to the stomach leaving the main vagal trunks along the distal 5 cm of esophagus in order to achieve both adequate control of acid secretion and also a lower duodenal ulcer recurrence rate. The data presented in this study of ten mongrel dogs suggest that, in the dog, division of the vagal fibers along the lesser curvature is more important in reducing acid secretion than is esophageal vagotomy; but the data also emphasize the contribution of the vagal fibers along the distal esophagus since a marked reduction in 2 DG-stimulated acid secretion can only be achieved by dividing the vagal fibers around the distal esophagus as well as those along the lesser curvature.


Presidential address: Gastrointestinal cancer. Surgical survey of abdominal tragedy. The experience with gastrointestinal cancer at a single large hospital has been utilized as a "micro-model" of the experience for the country at large, and the reasons why such a comparison might be possible have been presented. The incidence of various lesions has been discussed in terms of sex, age, and race. The changing incidence of various lesions has been pointed out. Survival results have been determined for total series, for various types of operative procedures, and for various extents of disease. The comparability of survival results from this one institution and those collected from the literature have been discussed. The emergence of newer diagnostic and therapeutic measures has been highlighted. The importance of education of both patients and physicians is emphasized repeatedly by the late stage at which so many of the patients with any gastrointestinal cancer present to and are diagnosed by the physician. The educational task for all of medicine is apparent and must be faced in some effective fashion.


The effect of feeding on the bile salt-independent canalicular secretion in dogs. Mongrel dogs were prepared by cholecystectomy, ligation of the lesser pancreatic duct, and insertion of cannulas into the duodenum and stomach. During experiments on the unanesthetized dogs, bile was collected by cannulating the common bile duct through the duodenal cannula. Canalicular secretion was estimated by the biliary clearance of erythritol. Feeding caused increased bile flow, increased erythritol clearance, decreased bile salt concentration, and unchanged bile salt output. These findings indicate that feeding stimulated the bile salt-independent canalicular secretion.


The relationship of cholecystectomy and taurocholic acid feeding to bile composition and hepatocyte function in prairie dogs. The prairie dog was used as a model for human gallstone formation. Stones formed in the gallbladder of all animals on a lithogenic diet. Hepatic bile was nonlithogenic, whereas gallbladder bile promoted cholesterol precipitation. Addition of taurocholate to the diet reduced the number of stones and lithogenicity. Cholecystectomy resulted in an increased bile flow and reduced secretion of cholesterol in the animals on a high cholesterol diet. Reduction of cholesterol and bile acid synthesis by negative feedback was demonstrated in isolated hepatocyte culture. The shift of bile salt production to chenodeoxycholates on a high cholesterol intake was demonstrated both in vivo and in vitro. A theory of gallstone formation is presented which hypothesizes a defect in hepatocyte storage of cholesterol rather than bile acid synthesis as the primary effect, relegating the problems to one of a disease of lipid metabolism.


Urease as a contributing factor in ulcerative lesions of the colon. Urease immunization protects animals against the development of uremic colitis. This indicates that ammonia formed by bacterial urease is the causative factor in the breakdown of the colonic mucosa.


A shunt equation for estimating the splenic component of portal hypertension. A modification of the Berggren pulmonary shunt equation was derived for use in estimating the splenic component of portal hypertension. It was used in nine patients, six of whom had sufficient decreases in portal flow and pressure after splenic artery and coronary vein ligation. The average decrease in portal flow was 52%.


Betatron therapy for unresectable pancreatic cancer. A preliminary report. Nineteen-patients with unresectable but localized ductal adenocarcinoma of the pancreas were treated with definitive high dose, small volume, external beam radiotherapy with promising results. The tumors of all patients were histologically confirmed. A 45 MV betatron was used for radiation of the clipped tumor volume plus a 1 to 2 cm margin. Fourteen patients were treated by a three-field technic with the use of opposed lateral photon beams and an anterior mixed photon-electron beam. Five patients were treated with either a four-field "box" or a three-field photon technic. Tumor doses of 5,900 to 6,700 r were delivered over seven to nine weeks. The twelve month survival rate after diagnosis is 53%. This compares favorably with published reports of patients treated surgically for cure. Four patients are alive 18.5 to 64 months after diagnosis. Three are clinically free of disease. Treatment has been well tolerated and there have been no significant radiation complications.


Surgical management of patients with primary operable colorectal cancer and synchronous liver metastases. The surgical results were analyzed of 217 patients who had undergone operation for primary colorectal cancer and were found to have synchronous liver metastases. It is recommended that patients with primary operable colorectal cancer who are found to have synchronous liver metastases should at least have an adequate resection of the primary lesion. If the primary lesion is resectable for cure and there are solitary hepatic metastases, then excision of these seems warranted. In most cases this can be accomplished by simple excision or wedge resection, although occasional cases may require lobectomy.


Vagal influence on duodenal motor activity. The extrinsic and intrinsic vagal influences on duodenal motor activity in the anesthetized dog were investigated. The following conclusions were derived: (1) Electrical vagal stimulation elicits gastroduodenal contractions. (2) Poststimulatory contractions, or off responses, occur on the antrum and duodenum after termination of the stimulus. (3) Denervation of the extrinsic vagal nerves to the duodenum does not eliminate the above responses. (4) Denervation of the extrinsic nerves to the duodenum and transection of the gastroduodenal junction eliminates stimulated duodenal motility and the occurrence of off responses on the duodenum. Antral responses remain unaffected. (5) The motor innervation to the duodenum is composed of extrinsic vagal fibers and intrinsic vagal nerves which appear to first enter the antrum and then cross the gastroduodenal junction to innervate the duodenum.


Mitogenicity of cell wall fractions of Micropolyspora faeni. Cell wall fractions of Micropolyspora faeni, a causative agent of hypersensitivity pneumonitis, have been shown to possess non-specific mitogenic activity for guinea pig, rabbit and human lymphocytes. This activity was unrelated to endotoxin and may not be associated with peptidoglycan. This lymphocyte proliferation stimulus may play a role in the adjuvant properties of this organism and in the etiology of hypersensitivity lung disease.


Age of onset in a group of severely asthmatic children. The age of onset was examined retrospectively in a group of severely asthmatic children. The mean age of onset for males was 29.4 +/- 22.7 months and 30.4 +/- 27.0 months for females. The distribution of age of onset between males and females  was remarkably similar (X2 = 1.77, 0.6 less than p less than 0.9). The mean age of onset was similar in children not receiving corticosteroids compared to those children who were taking high and low dose corticosteroids. An early or late age of onset appeared independent of corticosteroid requirements. Children with five or more positive immediate skin tests had earlier age of onset and higher total serum IgE levels than those with four or fewer positive skin tests.


Standardization of early drug trials in allergic diseases (with reference to a trial of doxantrazole). Attention is drawn to the long wait in certain countries before new drugs and other therapies for the treatment of allergic diseases are approved by the local authorities, even when good trials with them have been carried out elsewhere. The main reasons for requiring repeat trials are poor original case selection and inadequate parameters of assessment, particularly environmental variations. The results of re-assessing patients who in the previous year had undergone a trial of a new chromoglycate-like drug (Doxantrazole) are used to illustrate the need to introduce standard basic requirements when new drugs and other agents are being tested in allergic diseases. A suggested format of such requirements is presented.


A study of trends in upper arm soft tissue sarcomas in the state of Connecticut following the introduction of alum-absorbed allergenic extract. The incidence of soft tissue sarcomas of the upper and lower limbs in Connecticut from 1935-1974 was studied to determine whether the incidence of upper limb sarcoma changed following the introduction of alum-adsorbed allergenic extracts in 1963. No changes were observed that could be related to the use of alum.


Theophylline bioavailability following oral administration of six sustained-release preparations. Plasma theophylline concentrations were measured in 20 adults after single doses of 8 mg/kg of each of six oral sustained-release theophylline preparations. Twelve subjects were studied in a multiple-dose trial. Comparison of bioavailability and absorption characteristics suggest usefulness of some of these products for the purpose of maintaining safe and effective blood levels with twice-a-day dosing.


Giant hemangiomatosis of the liver in infancy. Two case reports of hepatic hemangiomatosis in infancy are presented to clarify the definition of this condition and to review its therapy by administration of corticosteroids and by ligation of the hepatic artery.


Pathophysiology and management of congenital posterolateral diaphragmatic hernias. Twelve neonates with foramen of Bochdalek hernias were treated at the Medical College of Virginia Hospitals during a recent four-year period. The clinical material supports the concept that bilateral hypoplastic lungs are fundamental in producing a morbid vicious cycle with respiratory and metabolic acidosis, pulmonary hypertension, right to left ductal and parenchymal shunting, worsening of the hypoxia and acidosis, and progression of the cycle. Management of Bochdalek hernias before, during and after operation has been guided by pathophysiologic approach.


Limb salvage by extended profunda femoris revascularization. We would conclude from the present series that if there is an adequate descending branch of the profunda femoris artery (20--25 cm), revascularization of the profunda femoris artery is worthwhile even in the presence of marked distal ischemia. In the present series, early success was manifested by relief of rest pain, healing of ischemic ulcers, and elevated ankle pressures (86%). The late success rate was 73%. If the proximal revascularization fails, distal bypasses can be added at a later procedure.


Surgical management of hyperparathyroidism. Twenty-one patients with hyperpara-thyroidism were studied to determine the outcome of surgical treatment by a variety of surgeons using a variety of techniques. Primary surgical treatment was excision of an adenoma in 11 patients with primary hyperparathyroidism. One patient (7%) had a true recurrence. One patient (7%) had persistent disease. Subtotal parathyroidectomies were carried out in three patients with primary diffuse hyperplasia and in five patients with chronic renal disease. Total parathyroidectomy and autotransplantation were performed in two more recent patients with chronic renal disease. Permanent hypoparathyroidism was not seen postoperatively in patients with primary hyperparathyroidism. Identification of all four glands at the operating table is essential. The low recurrence rate following selective excision of diseases parathyroid glands in this series suggests that the approach can be undertaken safely in most instances.


Response of first order vestibular neurons to lidocaine hydrochloride. Changes in behavior and electrical activity of primary vestibular neurons were observed following injection of lidocaine hydrochloride into the middle ear of cats. After injection the cats exhibited head and ocular nystagmus, head and neck deviation and pupillary changes. Mean preinjection resting discharge rate for first order vestibular neurons was 46.0 spikes/sec. Two hours after lidocaine application the resting rate decreased to a mean of 22.2 spikes/sec and then recovered to 43.0 spikes/sec four hours, and 47.4 spikes/sec six hours after the experimental treatment. The increment sensitivity (increased rate of firing) of horizontal canal neurons to constant angular acceleration for the control period was 2.0 extra spikes/sec/deg/sec2; two hours after the application, 70% of the recorded neurons were unresponsive to angular acceleration or tilt. The sensitivity after four hours was 1.0 spikes/sec/deg/sec2 and 2.1 spikes/sec/deg/sec2 at six hours. The distinct depression of sensitivity by lidocaine at four hours compared to normal mean resting rate at this time suggests these functions may be governed by two modes of action in the receptor or first order afferents.


Effects of combinations of sodium salicylate and noise on the auditory threshold. Thirty-nine monaural chinchillas were used to study the interaction between sodium salicylate and various TTS-producing noise paradigms. Five animals were included in each of the following three groups: 1) sodium salicylate (400 mg/kg) plus 2-4 kHz, 95 dB SPL noise band for one hour; 2) sodium salicylate (400 mg/kg) plus 4 kHz CF octave band noise at 80 dB SPL for 96 hours; and 3) sodium salicylate (400 mg/kg) plus 50 impulses having 50 musec A-duration and 158 dB peak SPL, presented at one per minute. The remaining 24 animals served as various controls in groups exposed to sodium salicylate or the noise paradigm alone. Thresholds were estimated before, during, and after exposure using the AER technique and cochleagrams were mapped for each cochlea 30 days after exposure. When sodium salicylate was combined with the various noise conditions, the maximum TTS values obtained from the combination studies were comparable to those obtained from the single agent producing the greatest TTS at a particular frequency. No consistent alteration in either magnitude or time course of posttreatment threshold shift was found following the combination treatments as compared to the individual agents alone. Hazard to the auditory system resulting from a combination of sodium salicylate and noise was concluded to be no greater than the hazard presented by either agent alone. This result is also substantiated in the histological results.


Bilateral acoustic neurinomas. A case report. A patient presented with complaint of increasing hearing loss in the right ear of five months duration. There was no hearing in the left ear as a result of previous surgery for resection of an acoustic neurinoma. Polytomes demonstrated a progressive erosion of the right auditory meatus. The patient underwent a right suboccipital craniotomy resulting in preservation hearing at a 30 dB level with 100% discrimination scores.


Current concepts of tumor immunology. III. Immune therapy. It is apparent that development of consistently effective methods of immunotherapy must await a more thorough understanding of the immune response to cancer. However, even those forms of immunotherapy which have been developed to date indicate a tremendous potential. It appears that immunotherapy may be most useful as an adjuvant to established forms of treatment. Surgery, radiation therapy and/or chemotherapy are used to remove all of the gross tumor, with immune therapy then employed to destroy the small foci of tumor which remain. As methods are developed which are effective in counteracting the immunosuppression of tumors, other means of immunotherapy may be found which are capable of destroying tumor cells while not affecting the adjacent normal tissue. Thus, the future of immune therapy holds great promise. As more is learned about the immune response to cancer, advances in therapy will certainly follow.


Treatment of infantile subglottic hemangioma with radioactive gold grain. Congenital subglottic hemangioma causes life-threatening airway obstruction during the first few months of life. The mortality rate of recognized and untreated cases justifies active treatment, and although radiation therapy is currently most favored, it carries a risk of inducing malignant change in the thyroid gland later in life. A series of 11 patients with laryngeal hemangiomata is reported, conventional radiotherapy was utilized in the first seven patients, and placement of a radioactive gold grain directly into the lesion was used in the last four patients. This technique offers maximal tumor dose with minimal thyroid gland irradiation compared to treatment by conventional radiotherapy, and its successful use in these four patients is reported as worthy of further trial.


Treatment of sudden deafness with sodium salts of triiodobenzoic acid derivatives. Twenty-two patients with a complaint of sudden deafness were treated with one of the other of two radiopaque contrast media, 9 received daily doses of sodium iothalamate, and 13 were treated similarly with methylglucamine and sodium iodamide. Four of the first group and eight of the second demonstrated either an effective partial or a complete recovery of hearing function. Those failing to respond were generally classified completely deaf at admission and report nausea, vomiting, or vertigo at onset of sudden deafness. Treatment with sodium salts of triiodobenzoic acid derivatives, i.e., sodium iothalamate and methylglucamine and sodium iodamide, is judged to be effective in selected cases of sudden deafness.


Studies on cochlear aqueduct patency.

Studies on cochlear aqueduct patency. Complex studies on the patency of the cochlear aqueduct were made on 250 human cadavers and the temporal bones isolated from these, using a chemical method (staining reaction) as well as filling the aqueduct with fluid plastics and exposing the whole of its course under an operating microscope. The disappearance of patency in the cochlear aqueduct is a progressive phenomenon reflecting the biological process of aging in the organism. Patency of the cochlear aqueduct renders possible a two-way spread of infection from the cerebrospinal fluid (CSF) to the inner ear and vice versa, CSF otorrhea, and sudden sensorineural hearing inpairment through rises in CSF pressure, while in the absence of patency, the accumulation of harmful products of metabolism in the perilymph has a deleterious effect on the sensory elements of the inner ear.


Critical look at the classification of maxillary sinus carcinomata. All the classification systems at present adopted for use in carcinoma of the maxillary sinus suffer from both intrinsic inaccuracies and an apparent failure to relate T (extent of primary tumor) categories to clinical experience of the spread of these tumors. Three systems are critically analyzed in relation to personal experience of 86 patients with sinus carcinoma and a fourth system proposed as a compromise solution to this problem.


Drug-related gustatory disorders.

Drug-related gustatory disorders. A review is presented of the drug-induced gustatory disorders that have so far been identified. Metallic, bitter or salty phantogeusias and hypogeusias, often of a dissociated type, which may even develop into ageusia have been described as side effects of treatment with some drugs. Personal observations on six cases are reported in which the gustatory disorders are attributed to the ingestion of phenylbutazone, carbamazepine, chlormezanone and baclofen. After interruption of the treatments these disorders were reversible within weeks to months.


Thermal injury from mastoid bone burrs.

Thermal injury from mastoid bone burrs. Marked bone temperature elevations and surface osteocyte necrosis result from cutting and diamond burrs used without continuous irrigation. Temperatures generated may injure the facial nerve even if drilling is carried on a millimeter or more distant from the nerve. Irrigation of the operative field maintains bone and nerve temperatures at safe levels. Cutting and diamond burrs obey different thermodynamic principles. Higher rotation speeds, greater applied pressures and greater surface contact area increase cutting burr thermogenesis. Burrs with more flutes generate greater heat than those with fewer flutes. Diamond burr heat generation varies in proportion to applied pressure, but is independent of rotation speed or contact area. Haversian canal blood vessels do not contribute to postoperative revascularization. Rather, vessels immigrate from adjacent intact periosteum, penetrating arterioles and exposed marrow. Surface thermal injury does not impede revascularization and should not impair the healing of tissue grafts applied to the bone surface.


Blocking effect of radio-contrast media on cochlear depression. Furosemide injection was used in a total of 72 guinea pigs to induce a cochlear dysfunction similar to the sensorineural loss associated with sudden deafness. Prior to the furosemide administration, 38 of the animals were treated with an injection of one of four radio-contrast media; methylglucamine and sodium diatrizoate, sodium iothalamate, methylglucamine and sodium iodamide, and methylglucamine diatrizoate. In the control animals, receiving only furosemide, both endocochlear potential (EP) and cochlear microphonic (CM) underwent a severe depression followed by recovery and subsequent secondary depression. Animals pretreated with one of the first three compounds showed little or no EP or CM depression. Animals pretreated with the fourth, methylglucamine diatrizoate, showed the same EP depression as the control animals. The possible mechanism for this blocking effect and its significance in the clinical treatment of sudden deafness is discussed.


Endobronchial foreign body removal using the bronchofiberscope. In certain situations the flexible fiberoptic bronchoscope can augment rigid bronchoscopy in foreign body removal. A case of a successful fiberoptic removal of an endobronchial foreign body (dental bridge) is presented. A wire basket, inserted through the channel of the bronchofiberscope, was used to capture the object.


Airway management in the premature infant. Out of 262 premature newborn patients admitted with a diagnosis of respiratory distress, it was necessary to treat 70 with a ventilator. Of these 70, 25 eventually underwent tracheostomy. Indications for tracheostomy were that of an infant needing prolonged endotracheal intubation greater than one week. The procedure itself was easily performed and an overall complication rate of 7% was the result. Of the patients who underwent tracheostomy, 8% had significant complications. There was no death attributable to the treatment regime. We feel, therefore, that a combination approach starting with the endotracheal tube and progressing to tracheostomy when necessary, provided the best care for premature infants requiring intensive airway management.


Primary malignant lymphoma of the paranasal sinuses. A case of primary malignant lymphoma of the paranasal sinuses is presented. The patient, a 56-year-old white man, had a good response to Cobalt 60 therapy. A review of the literature reveals that this rare tumor occurs predominantly in older individuals, and that the chance for survival is good when radiation has been administered.


Nasopulmonary reflex: evaluation in the nonparalyzed and paralyzed anesthetized dog. Changes in the pulmonary mechanics of various animal species have been observed after nasal stimulation. In reviewing the methods of these studies, we found that often the respiratory patterns were not controlled well enough to permit conclusions about changes in resistance and compliance. To further investigate the possibility of a nasopulmonary reflex, we first studied the effect of nasal stimulation on pulmonary mechanics in anesthetized, nonparalyzed dogs. In this group of 35 dogs, we found that nasal stimulation typically produced marked changes in breathing patterns, a large transient increase (150%) in pulmonary airflow resistance, but no change in compliance. This response to nasal stimulation could be abolished by interrupting either the trigeminal or the vagus nerves. In an additional group of 12 anesthetized, paralyzed dogs, we found no changes in resistance or compliance after nasal stimulation. From this observation, we concluded that nasal stimulation produced changes in breathing patterns that led to alterations in resistance without an actual change in the intrinsic behavior of the pulmonary airways.


Comparison of five types of tracheostomy tubes in the intubated trachea. Recognition of excessive cuff pressures as a major factor in tracheal injury has prompted the development of different types of cuffed tracheostomy tubes. Five brands of tracheostomy tubes in common use are evaluated in anesthetized adult dogs. Visualization of interior tracheal conditions during spontaneous and positive pressure ventilation permits a dynamic study of the trachea and its interaction with each tube under varying conditions. The relationship between intracuff pressure and cuff-tracheal wall pressure is analyzed for each tube. The selection of the proper tracheostomy tube for a given clinical situation should consider: 1) the cuff pressure necessary to achieve minimal leak, 2) torque transmission, 3) the ability to monitor cuff-tracheal wall pressure as a function of intracuff pressure, and 4) simplicity of design for optimum clinical use. Tracheal expansion occurs in a predictable fashion in response to different stimuli. However, the role of expansion in the susceptibility of this organ to injury remains to be investigated.


Idiopathic midface lesions.

Idiopathic midface lesions. The term lethal midline granuloma has been used to describe a number of lesions which may present in the midface. A list of possible causes is presented and three entities, midline malignant reticulosis, Wegener's granulomatosis, and malignant lymphoma, are discussed in detail. The pathological descriptions, the clinical features, and the recommended forms of therapy are outlined. It is important to vigorously pursue the patient with a midline facial lesion until a definite tissue diagnosis is obtained. Multiple biopsies may be necessary because necrosis and nonspecific inflammatory changes may be present in various of these disease entities. The correct diagnosis is essential because the treatment of these disease is different: Wegener's granulomatosis responds best to chemotherapy, whereas midline malignant reticulosis and lymphoma should be treated with radiation therapy if they are localized. Despite conceptual variations, most recent authors believe that these three diseases exist as separate entities, and furthermore, recommend that the term lethal midline granuloma be either dropped from the lexicon or relegated to a descriptive clinical term to be used only until a more definitive diagnosis can be made.


Glutathione reductase in the red blood cells. Glutathione reductase plays an important role in protecting hemoglobin, red cell enzymes, and biological cell membranes against oxidative damage by increasing the level of reduced glutathone (GSSGR) in the process of aerobic glycolysis. The enzyme deficiency may result in mild to moderately severe hemolytic anemia upon exposure to certain drugs or chemicals. However, hereditary deficiency of the enzyme is extremely rare. Recent studies on glutathione reductase in the red cell have shown more insight in the understanding of red cell metabolism and interactions with other enzymes, especially glucose-6-phosphate dehydrogenase (G-6-PD). Glutathione reducatase in serum may be a source of error in any clinical laboratory test in which an enzyme activity is determined indirectly by measuring the change in reduced nicotinamide-adenine dinucleotide (NADH) or reduced nicotinamide adenine dinucleotide phosphate (NADPH) absorbance. Glutathione reductase levels are reduced in banked blood when citrate-phosphate-dextrose (CPD) is used as a preservative. Reviewed is the role of glutathione reductase in the metabolism of the red cell and its clinical implication and usefulness.


Otosclerotic inner ear syndrome.

Otosclerotic inner ear syndrome. Twenty-seven patients with clinical otosclerosis and vertigo underwent stapedectomies. Vertigo improved or completely disappeared in 23 patients, indicating that stapedectomy is probably helpful. Three case reports are also presented as well as photomicrographs of temporal bones showing the focus in contact with the pars superior.


[Estimation of the heritability of hair and eye color]. In the present study the colours of hair are regarded as quantitative genetic characters. The genetic and environmental components of the total variance of the characters are determined by analysis of correlation between parents and children as well as full sibs. An analysis of correlation coefficients makes it possible to further examine whether autosomal dominance or X-linked factors play a role in the inheritance. The correlations between the parents make the genetical analysis more complicated as they raise the correlation coefficients between relatives. The estimates of heritability are 0.61 (hair-colour) and 0.80 (eye-colour). These estimates may serve as preliminary values of orientation. Only further studies will show whether the relative large part of the variance caused by the environment can be confirmed. Dominance may play a small role in the case of hair colour, while it seems to be absent in eye-colour. There are no sufficient indications of X-linked factors for both characters.


Current status of carcinoembryonic antigen (CEA) and CEA-S assays in the evaluation of neoplasm of the gastrointestinal tract. Carcinoembryonic antigen (CEA) is heterogeneous and may represent a set of closely related glycoproteins which have been referred to as isomeric species of CEA. The existence of other CEA-related glycoproteins such as non-specific cross reacting antigen (NCA) has been recognized. Recently a highly purified homogeneous isomeric species of CEA described as CEA-S has shown differences in diagnostic results upon analysis of clinical sera as well as quantitative immunochemical differences. In a blind study of 308 sera, the CEA and CEA-S assays were compared. A significant difference in false positive results was observed between the CEA and CEA-S assay results. In contrast to the low but significant evidence of elevated CEA in sera of random normal persons and patients with liver disease or inflammatory disease of the gastrointestinal tract or lung, none of the sera had elevated concentrations of CEA-S. Among patients with tumor of the gastrointestinal tract that were considered surgically resectable, 46 percent were elevated using the CEA-S assay and only 34.7 percent were elevated above 5 nanograms per ml by the CEA assay. The CEA assays detect CEA-related molecules produced by lung, breast and other tumors; the CEA-S assay appears equally sensitive to CEA of gastrointestinal origin but detects only a small subgroup of breast, lung and, rarely, other types of tumors.


Detergent altered alkaline phosphatase patterns of liver disease. A procedure has been described whereby the high molecular weight alkaline phosphatase (slow-moving) isoenzyme may be studied by means of polyacrylamide gel electrophoresis. By treatment of sera containing this isoenzyme with some detergents of the nonionic Triton octylphenoxyethanol series, the high molecular weight alkaline phosphatase isoenzyme is altered so that its electrophoretic mobility more closely resembles that of the usual alklaine phosphatase isoenzymes. The high molecular weight isoenzyme is thought to be associated with phosphatidyl choline and/or liproproteins. The detergent action is to dissociated the alkaline phosphatase from its lipid carrier. It is thought that these lipid-alkaline phosphatase complexes are associated with liver cell fragments. The detergent altered slow-moving alkaline phosphatase may migrate as a single band, from two to four new bands, or as several new bands. Liver, bone and intestinal alkaline phosphatase isoenzymes are unaffected by detergent action.


Serum protein polymorphisms Hp, Tf, Gc, Gm, Inv and Pt in Bantu speaking South African Negroids. Frequencies for the genetic polymorphisms of the serum proteins Hp, Tf, Gc, Gm, Inv and Pt are presented for 7 ethnically defined Negroid sub-samples (total n = 918--977, depending on genetic marker system) from South Africa. The results are discussed in terms of previous findings and of factors assumed to have an influence on the distribution of the particular genetic markers. The present report on the Pt ("post transferrin" approximately C3) polymorphism represents the first report on the genetic variation of the corresponding beta1A-globulin fraction within African Negroids (PtA = 0.009, PtB = 0.966, PtC = 0.025).


Chemotherapy of human tumor xenografts in genetically athymic mice. A series of studies were undertaken to evaluate the chemotherapeutic response to various antineoplastic drugs of human breast (MX-1) or colon (CX-2) tumor xenografts growing in genetically athymic (nude) mice. Fragments (2mm3) of either tumor type were implanted subcutaneously into the subaxillary region of NIH Swiss nude mice, and single drug therapy was started when tumors became palpable and were growing progressively. 5-Fluorouracil (5-FU) administered on a Q7DX3 schedule starting on Day 21 post tumor implantation elicited significant retardation in the growth rate of CX-2 tumor. A single treatment with methyl-CCNU induced temporary tumor regression. Against MX-1 tumor, both cyclophosphamide and melphalan induced tumor regressions with no recurrence. 5-FU slowed but did not arrest growth of MX-1 tumor. These tumor systems grown in nude mice appear to be suitable models for in vivo screening of anticancer agents that would prove clinically active.


The weak binding reaction between folate and human serum proteins. New evidence is presented that folic acid in serum binds with low affinity to major serum proteins. This low affinity binding is distinct from the high affinity binding by folate binding protein (FBP), a minor protein which is known to occur in serum with great quantitative variability. These conclusions are based on results obtained by equilibrium dialysis of serum containing only negligible amounts of FBP. At 4 degrees, the equilibrium value of the ratio of bound to free folate was approximately 0.81 and remained the same even wth up to 1,000 times greater than normal folic acid concentrations; however, with higher concentrations the ratio decreased progressively. These results are predicted by the laws of mass equilibria for a binding system with low ligand concentration vis-a-vis high binder concentration and low affinity between the reactants. A rough estimate of the mean affinity constant K governing this weak folate interaction with serum proteins yielded a value of 1.12 +/- 0.13 X 10(3) M-1.


Estimation of bilirubin binding capacity of neonatal serum. One aim of laboratory investigation in neonatal jaundice is to prevent kernicterus from occurring. Estimation of the bilirubin-binding capacity of serum is an important measurement along with total bilirubin determination. With a convenient Sephadex competitive binding procedure, KERNLUTE provides information on whether or not the neonate's bilirubin-binding capacity has been exceeded. Further testing of those serum specimens which show some residual binding capacity estimates whether the remaining bilirubin capacity is equivalent to 0 to 2.5 mg, 2.5 to 5 mg or more than 5 mg bilirubin per dl. References on the utility of this method and other more cumbersome procedures are given.


[Role of cobalt in the biosynthesis of the components of the gentamicin complex]. It was shown that addition of cobalt ions or vitamin B12 to the fermentation medium resulted in an increase in the level of gentamicin accumulation, the relative content of the most methylated components C1 and C2 in the gentamicin complex being increased, while the content of the least methylated components CIa and "minors" decreased. Addition of sulphodimezine lowered the gentamicin biosynthesis rate and the relative content of gentamicins C1 and C2. It was supposed that cobalt stimulated the B12-dependent synthesis of methionine, being the source of the methyl groups for biosynthesis of the methylated components of gentamicin complex.


[Selection of active strains of Actinomadura carminata, the producer of carminomycin]. The method of selecting active strains among a definite group of analogue-resistant mutants was used on the basis of studying the dependence of the carminomycin-producing organism growth and the antibiotic synthesis level on some metabolites. As a result, gamma-ray induced mutants 4 times more active than the parent strains were obtained.


[Study of the antibiotic parvulomycin. The isolation of alpha,alpha-trehalose and L-glutamic acid]. A non-reducing disugar and amino acid were isolated in the studies on the structure of parvulomycin. The acid hydrolysis of the disugar revealed the presence of 2 moles of D-glucose. Acetylation of the disugar resulted in formation of octa-O-acetyl-alpha,alpha-tregalose, saponification of which resulted in formation of alpha,alpha-tregalose. Its physical parameters, i.e. melting point of the mixed sample, optical rotation, IR-spectrum coincided with those of the authentic alpha,alpha-tregalose. The isolated amino acid proved to be L-glutamic acid on thin-layer chromatography with witness and comparison of the physico-chemical properties of their hydrochlorides.


[Biosynthesis of the labelled rifamycin B in the presence of different 14C- and 3H-precursors]. The results of studying the effectiveness of incorporation of the label from different 14C- and 3H-precursors into the molecule of rifamicin B during its biosynthesis are presented. The regularities of the label incorporation into the antibiotic composition as dependent on the time of the precursor addition were investigated. A radiochemically pure preparation of 14C-rifamicin B with specific radioactivity of 3 mcurie/mg was obtained.


[Development of a method for the quantitative assessment of microorganism sensitivity to antibiotics using discs. The study of different nutrient media for determining microorganism sensitivity to antibiotics]. Five nutrient media used for determination of microbial sensitivity to antibiotics, i.e. beaf-peptone agar, Hottinger pancreatic beaf infusion agar, sprat hydrolysate nutrient agar of the Dagestan Research Institute of Nutrient Media, Muller-Chintone agar from Bulgaria and "Oxoid" agar for determination of microbial sensitivity were studied comparatively. The media were compared with respect to the growth density with the use of different test-cultures and the clearance of the inhibition growth zones around the discs containing different antibiotics. The best results were obtained with the use of sprat hydrolysate nutrient agar. Further studies on the medium standardization are necessary.


[Properties and stability of tetracycline hydrochloride tablets]. Three forms of tetracycline hydrochloride tablets were studied, i.e. the uncoated tablets, sugar-coated tablets and the tablets with gastric-soluble film coating. The film-coated tablets had advantages in the main quality characteristics, i.e. the contents of the tetracycline-like admixtures, the active substance and solubility rate. The possibility of the tablet stability investigation by the method of "accelerated aging" was shown.


[Effect of triterpene glycosides on the plasma membrane permeability for amino acids in Saccharomyces carlsbergensis yeast cells]. The effect of triterpen glycosides, such as cauloside C from Caulophyllum robustum, stichoposide A from Stichopus japonicus S and the asaponine from Thea sinensis L on permeability of the plasmic membranes for amino acids was studied. It was shown that the glycosides induced higher levels of liberation of the amino acids with positively or negatively charged molecules from the cells of Saccharomyces carlsbergensis as compared to the control. The transport of the non-polar amino acids was least affected by addition of the saponines to the incubation medium.


[Formation by the mycoheptin producer of aureothin and pentaene 2814-P, separated into its mycopentene-1 and mycopentene-2 components]. Aureothin, a toxic substance of non-polyenic nature and pentaen 2814-P were isolated from the metabolites of Streptoverticillium mycoheptinicum, the organism producing mycoheptaine. The above substances are also produced by other actinomycetes. The pentaenic antibiotic was divided into 2 components. Physico-chemical and biological properties, as well as the composition of the substances were studied.


[Effect of therapeutic doses of tetracycline on the structure of the thymus gland in rabbits]. Tetracycline was administered orally to Shinshilla male rabbits (female, 2.0-2.5 kg) twice a day, the daily dose being 20 mg. The thymus was examined in 1 and 2 weeks by the methods of histo- and cytometry. The reliability of the differences and correlation were estimated by the non-parametric methods. Out of 15 parameters studied 8 showed reliable differences and 5 were statistically connected. The following structural shifts were found: (1) increased numbers of activated and middle thymocytes evident of intraorgan cell activation, (2) higher mitotic index and lower index of the cell degradation resulting in the cortex hypertrophia by the end of the week, (3) increased cell density in the medullar substance which is the morphological criterion of intraorgan migration of the cells from the cortex into the medullar substance. Thus, the morphometric analysis of the thymus proved to be much more informative than the usual morphological analysis. The observed structural change in the thymus is considered to be a result of the antigenic effect of tetracycline on the host.


[Comparative study of the cardiotoxicity of the anthracycline antibiotics, rubomycin, carminomycin and dihydrocarminomycin, in experiments on white mice]. The experiments on albino mice treated with rubomycin, carminomycin or dihydrocarminomycin on its 5-fold intravenous administration in doses equal to similar portions of LD50 of the respective antibiotic on its use in a single dose showed that all the 3 antibiotics induced changes in the myocardium close by their character. The heart affections were evident from swelling of separate muscle fibers, degeneration of the myofibrils, homogenization, vacuolization and resorption of the sarcoplasma, pathological changes in the nuclei, atrophy of some muscle fibers. Rubomycin had the highest cardiotoxic effect. Then followed dihydrocarminomycin and carminomycin. All the antibiotics studied in the experiments with mice had mainly an inhibitory effect on the lymphoid hemopoiesis. The effect of carminomycin was the highest. The animal death during the injections and immediately after administrations of the antibiotics must be due to their suppressing effect on hemopoiesis. The deaths at more remoted periods must be due to the cardiotoxic effect of the antibiotics.


Reliable sampling method for analysis of the ecology of the human alimentary tract. An intubation method has been developed that allows removal of a sample of human intestinal fluid within a short period of time, that avoids contamination, and that minimizes exposure of the sample to air. Preliminary results obtained with this method have shown that the stomach and duodenum are essentially sterile and that the bacterial population present in the remainder of the small intestine is similar to that described by previous workers except that Veillonella species were encountered frequently and Haemophilus species were also detected in the lower jejunum and ileum of some individuals.


Upper boundary of the biosphere.

Upper boundary of the biosphere. By using meterological rockets fitted with specially designed analyzers, samples for microbiological investigation have been taken. The analyzer design prevented extraneous microorganisms from penetrating into the analyzer. Before being used, the analyzers were sterilized with high gamma-ray doses. For the first time microorganisms have been detected in the mesosphere at an altitude of 48 to 77 km. The microorganisms are microscopic fungi having black conidia or spores (Circinella muscae, Aspergillus niger, Papulaspora anomala) and one species forming green conidia (Penicillium notatum). Colonies of Mycobacterium luteum and Micrococcus albus have also grown. Five of the six species have synthesized pigments. The presence of pigmented microbial forms leads us to believe that natural selection is occurring in the mesosphere because cells possessing chromogenous pigments (carotenoids, melanins) are more resistant to ultraviolet-ray action. A greater number of microorganisms have been registered in the mesosphere during dust storms than in the absence of strong winds.


Wild rice as fermentation substrate for mycotoxin production. Many cereal grains have been studied for their suitability as substrates for the fermentative production of mycotoxins. However, except for aflatoxin, wild rice has not been investigated. Hence, five mold cultures known to produce the mycotoxins ochratoxin-A, penicillic acid, patulin, vomitoxin, and zearalenone were grown on wild rice under varying conditions of moisture and temperature to determine whether this grain would serve as a suitable substrate for toxin production. Under appropriate fermentation conditions, good yields of ochratoxin-A and moderate amounts of patulin were obtained, but only small amounts of penicillic acid, vomitoxin, and zearalenone were elaborated. An extract from a sample of naturally molded wild rice contained 0.8 microgram of patulin per g of rice. The predominating mold was identified as Aspergillus clavatus. Under identical cultural conditions, this isolate and a known patulin-producing strain of A. clavatus yielded approximately equivalent amounts of the mycotoxin.


Toxicity of Bacillus thuringiensis spores to the tobacco hornworm, Manduca sexta. Toxicity of Bacillus thuringiensis spores to the tobacco hornworm, Manduca sexta, is described. The numbers of larvae killed were in relation to spore dry weight. At a surface application of 6.8 ng/cm2, there was an 85 percent survival, but less than 50 percent survived at 68.2 ng/cm2. Striking similarity of spores to parasporal crystals is revealed by slope of mortality curves, inhibition of stadial growth, and 50 percent lethal dose values based on protein content.


Nitrogen fixation (acetylene reduction) associated with roots of winter wheat and sorghum in Nebraska. Root segments and root-soil cores (6.5-cm diameter) from fields and nurseries of winter wheat and sorghum were tested for N2 fixation by using the acetylene reduction assay. Wheat samples (approximately 1,200) from 109 sites generally had low or no activity (0 to 3.1 nmol of C2H4 produced per h per g [dry weight] of root segments), even after 24 h of incubation. However, a commercial field of Scout 66, located in western Nebraska, exhibited appreciable activity (290 nmol of C2H4 produced per h per g [dry weight] of root segments). Of 400 sorghum lines and crosses, grain sorghums (i.e., CK-60A, Wheatland A, B517, and NP-16) generally exhibited higher nitrogenase activity than forage sorghums or winter wheats. CK-60A, a male sterile grain sorghum, was sampled at four locations and had the most consistent activity of 24 to 1,100 nmol of C2H4 produced per h per core. The maximum rate extrapolated to 2.5 g of N per hectare per day. Numerous N2-fixing bacterial isolates were obtained from wheat and sorghum roots that exhibited high nitrogenase activity. Most isolates were members of the Enterobacteriacae, i.e., Klebsiella pneumoniae, Enterobacter cloacae, and Erwinia herbicola.


Adhesion of bacteria to epithelial cell surfaces within the reticulo-rumen of cattle. Blocks of tissue were removed from various locations in the bovine digestive tract and fixed and processed for transmission and scanning electron microscopy by techniques that retained adherent bacteria. The distribution of bacteria on the surface of epithelial cells was examined by scanning electron microscopy. This showed intermittent colonization of the epithelia with the formation of occasional microcolonies of morphologically similar bacterial cells. Transmission electron microscopy of ruthenium red-stained material showed the presence of both the glycocalyx of the bovine epithelial cells and fibrous carbohydrate coats surrounding adherent bacteria. The carbohydrate coats appeared to mediate the attachment of bacteria to the epithelium, to food particles, and to each other so that microcolonies were formed. Careful examination of the bacterial colonization of keratinized cells in the process of being sloughed from the surface of the stratified squamous epithelium of the rumen showed that these dead cells were digested by adherent bacteria of a limited number of morphological types. The spatial relationship of this mixed, adherent, microbial population to living and dead epithelial cells and to food particles indicates that digestive processes of some importance may be accomplished by this stationary component of the microbial flora of the digestive tract.


Immunolatex localization by scanning electron microscopy of intestinal bacteria from cockroaches. An immunolatex reagent was developed from antiserum to Fusobacterium varium, an obligate anaerobe isolated from the colon of the cockroach, Periplaneta americana L. A new technique that enabled the preparation of a highly efficient immunolatex conjugate was used to localize the bacterium with scanning electron microscopy, in situ, in the mixed gut contents.


Analysis of bacterial fermentation products by isotachophoresis. Various carboxylic acids from bacterial fermentation could easily be separated by isotachophoresis. The analyses were performed on an LKB 2127 Tachophor, and under the conditions used the minimum amount of sample that could be quantitatively estimated was approximately 0.1 nmol. The reproducibility of the method was good (ca. 5 percent). The time of analysis using a 23-cm column was 12 min. No pretreatment of the samples was required.


Dulcitol-malonate-phenylalanine agar for the identification of Salmonella and other Enterobacteriaceae. An agar medium combining dulcitol fermentation, malonate utilization, and phenylalanine deamination was evaluated with 229 isolates representing 19 genera. All reactions agreed with those obtained on conventional media.


Isolation of bacteriophage 14-lysogenized Salmonella from the freshwater aquarium snail Apullaria. A naturally occurring Salmonella mikawasima serologically converted by phage 14 (6,7,14:y:e,n,z15) has been isolated for the first time. An S. tennessee variant seroconverted by phage 14 (6,7,14:z29:-) was also isolated. The source of these salmonellae was the common freshwater aquarium snail Ampullaria. Phage 14 prepared from these serovariants was lytic for S. bovis-morbificans (6,8:r:1,5) and for S. hadar (6,8:z10:e,n,x).


New sensitive biological assay for 12,13-epoxytrichothecenes. Germination of Nicotiana sylvestris pollen is inhibited by T2 toxin and diacetoxyscirpenol. Based on this observation, an assay method was developed that is capable of detecting nanogram quantities of these toxins.


Pectolytic enzymes in Rhizobium.

Pectolytic enzymes in Rhizobium. A sensitive pectin agar plate assay was used to demonstrate low levels of pectolytic enzymes in infective and noninfective strains of Rhizobium. The possible relation of this characteristic to legume infection is discussed.


Catechol and phenol degradation by a methanogenic population of bacteria. An anaerobic population of bacteria became acclimated to catechol and phenol in 32 and 18 days, respectively. Evidence from carbon balance measurements indicates that the aromatic ring is cleaved and that the products are stoichiometrically fermentable to methane and carbon dioxide.


Vibrio anguillarum and larval mortality in a California coastal shellfish hatchery. Vibrio anguillarum was isolated as a pathogen in the commercial culture of oyster spat at Pigeon Point, Calif. A water-soluble, heat-stable exotoxin extracted from cultures of the vibrio inhibited larval swimming and contributed to larval mortality. Although the vibrio was insensitive to penicillin in standard plate testing, this antibiotic proved useful in preventing mass larval mortalities in the hatchery.


Relative significance of exogenous  and de novo synthesized fatty acids in the formation of rumen microbial lipids in vitro. Mixed rumen microorganisms (MRM) or suspensions of rumen Holotrich protozoa obtained from a sheep were incubated anaerobically with [1-(14)C]linoleic acid, [U-(14)C]glucose, or [1-(14)C]acetate. With MRM, the total amount of fatty acids present did not change after incubation. An increase in fatty acids esterified into sterolesters (SE) and polar lipids at the expense of free fatty acids was observed. This effect was intensified by the addition of fermentable carbohydrate to the incubations. Radioactivity from [1-(14)C]linoleic acid was incorporated into SE and polar lipids with both MRM and Holotrich protozoa. With MRM the order of incorporation of radioactivity was as follows: SE > phosphatidylethanolamine > phosphatidylcholine. With Holotrich protozoa, the order of incorporation was phosphatidylcholine > phosphatidylethanolamine > SE. With MRM the radioactivity remaining in the free fatty acids and that incorporated into SE was mainly associated with saturated fatty acids, but a considerable part of the radioactivity in the polar lipids was associated with dienoic fatty acids. This effect of hydrogenation prior to incorporation was also noted with Holotrich protozoa but to a much lesser extent. Small amounts of radioactivity from [U-(14)C]glucose and [1-(14)C]acetate were incorporated into rumen microbial lipids. With protozoa incubated with [U-(14)C]glucose, the major part of incorporated radioactivity was present in the glycerol moiety of the lipids. From the amounts of lipid classes present, their radioactivity, and fatty acid composition, estimates were made of the amounts of higher fatty acids directly incorporated into microbial lipids and the amounts synthesized de novo from glucose or acetate. It is concluded that the amounts directly incorporated may be greater than the amounts synthesized de novo.


Role of heterotrophic nutrition in growth of the alga Scenedesmus obliquus in high-rate oxidation ponds. The green alga Scenedesmus obliquus readily adapted to heterotrophic growth in the dark, utilizing glucose as the sole carbon source. Heterotrophic cells differed significantly from photoautotrophic cells with respect to several physiological properties such as the rate of photoassimilation of CO2, rate of incorporation of glucose, and chlorophyll a concentration. Oxidation pond cells shared features common to both photoautotrophic and heterotrophic cells. Approximately 15 percent of oxidation pond algal carbon was derived from glucose assimilated directly without first being oxidized by bacteria. Bacteria seem to play a minor role in biological oxygen demand reduction in high-rate oxidation ponds, and their role is probably confined to degradation of biopolymers, thus producing substrates for algal consumption.


Phototrophic purple and green bacteria in a sewage treatment plant. In all purification stages of a biological sewage treatment plant, phototrophic bacteria were detected by the method of viable cell counts. The predominant species identified belonged to the genus Rhodopseudomonas of purple nonsulfur bacteria. The number of phototrophic bacteria was highest in wastewater containing sludge. In activated sludge, an average of 10(5) viable cells/ml was found; the number depended upon concentration of sludge rather than on seasonal changes in light conditions in the course of a year. Bacteriochlorophyll a was extracted from activated sludge. Relative to the viable counts of phototrophic bacteria, the content of bacteriochlorophyll a was 5- to 10-fold higher than that of three representative pure cultures. By incubation of activated and digester sludge under different environmental conditions, it was shown that phototrophic bacteria can complete with other bacteria only under anaerobic conditions in the light.


Discrimination between oral streptococci by pyrolysis gas-liquid chromatography. Washed organisms, including strains of Streptococcus mitior, S. mutans, and S. sanguis, were examined by curie-point pyrolysis gas-liquid chromatography. A linear discriminant function based upon three items from the output data was adequate for segregating the strains according to species. Strains with intermediate properties were also encountered. Sources of variability in cultures were evaluated, chromatographic performance was maintained throughout the investigation, and matching performance from a duplicate pair of chromatographic columns was demonstrated.


Structural factors influencing the biodegradation of imides. Comparative studies on the biodegradability of amides and imides are presented. Low-molecular-weight imides of varying chain lengths (4, 6, 7, 8, 18, and 20 carbons) were biodegrable. N-alkyl substitution of amides and imides resulted in non-biodegrable derivatives when the amide portion was greater than two carbons in length. N-alkyl-substituted derivatives of acetamide or diacetamide, however, were biodegrable. Several soil isolates, including Aspergillus niger and species of Flavobacterium and Alcaligenes, were capable of growth with imides as sole N or C sources.


Regional variations of cutaneous propionibacteria. Propionibacterium acnes, P. avidum, and P. granulosum were quantitatively measured in 50 young adults. The scalp, forehead, external auditory canal, alae nasi, anterior nares, groin, rectum, and antecubital and popliteal fossa were sampled. These represent various cutaneous microenvironments, differing in moisture, density of sweat, sebaceous glands, and extent of anaerobiosis. These studies show that the propionibacteria are ubiquitous on the skin, with P. acnes predominant in both prevalence and population, especially in areas rich in sebum. P. granulosum recovery paralled that of P. acnes, but the density was significantly lower. P. avidum was found mainly in moist areas and the retum, suggesting an intestinal reservoir.


Distribution of autotrophic nitrifying bacteria in a polluted river (the Passaic). The abundance of nitrifying bacteria, determined by most-probable-number procedures, within habitats of the Passaic River was as follows: rooted aquatic plants greater than algae approximately equal to rocks greater than sediments greater than greater than water. On the average, NH4+ oxidizers were 540-fold more abundant in the topmost 1 cm of sediment than in the water, and NO2- oxidizers were 250-fold more abundant. The population densities in this surface sediment at two nearby stations, one with a predominantly mineral stream bed and the other an organic ooze, did not differ significantly. Large numbers of nitrifiers were present to a depth of about 5 cm in a mineral sediment core.


Compact liquid nitrogen storage system yielding high recoveries of gram-negative anaerobes. A simple and compact system suitable for the preservation of fragile gram negative anaerobes and other bacteria in liquid N2 has been developed. Polypropylene straws used as specimen containers can be used easily within glove bags of anaerobic chambers, and their small size greatly increases the number of cultures which can be stored. Ancillary equipment and methods developed are described. The overall system was tested, using Streptococcus mutans, Fusobacterium nucleatum, and Selenomonas sputigena. Various basal suspending fluids and cryoprotective supplements were studied. With fast rates of freezing and thawing, survival recoveries of the test microorganisms ranged from 80 to 100 percent of the input colony-forming units in a complex medium broth base without cryoprotective agent addition, and they consistently were 100 percent when 0.4 mM polyvinylpyrrolidine was used. Overall, cryoprotection by polyvinyl pyrrolidine was superior to that from glycerol or dimethyl sulfoxide, the latter yielding recoveries similar to or less than those obtained with no cryoprotectant additive. All microorganisms were recoverable after storage for 1 year.


Usefulness of the stomacher in a microbiological regulatory laboratory. The relative efficiency of the Waring blender, the Stomacher 400, and the Stomacher 3500 for preparing food samples for microbiological analysis was studied. Comparative aerobic plate count (APC) values were determined on 671 samples, representing 30 categories of foods. Of the 26 categories of nonfatty foods, the blender gave significantly higher geometric mean APC values than those given by the Stomacher 400 and the Stomacher 3500 in 65 and 69 percent of the categories, respectively. In a comparison of the two Stomacher models, the Stomacher 400 gave significantly higher geometric mean APC values than these given by the Stomacher 3500 in 73 percent of the food categories. Addition of Tween 80 to four categories of fatty foods at concentrations of 0.5, 1.0, and 2.0 percent did not raise the APC values given by either model of stomacher to the levels given by the Waring blender. Overall, the efficiency of both models of Stomacher, relative to the blender and to each other, was specific and depended upon the particular food being analyzed.


Differentiation of Salmonella Serotypes by pyrolysis-gas-liquid chromatography of cell fragments. Pyrolysis-gas-liquid chromatography (PGLC) of whole cells and cell fragments was used to differentiate 10 Salmonella serotypes. Lyophilized samples (200 microgram) of whole cells, cell walls, flagella, and deoxyribonucleic acid from each serotype were analyzed in duplicate by PGLC. Pyrochromatograms recorded as pyrolytic elution patterns represented thermal fragmentation products of the samples. Mathematical expressions of percent similarity and percent conformity were calculated for all possible pair combinations of the 10 serotypes. Stepwise discriminant analysis of the PGLC data showed that 100 percent correct classification of the 10 serotypes was possible from the flagella or deoxyribonucleic acid pyrochromatograms. The classification matrix of the whole-cell data showed a 90 percent correct classification. PGLC of cell fragments may provide useful information for taxonomic studies of Salmonella and other microorganisms.


Physiological and biochemical contributions to the taxonomy of the genus Prototheca. II. Starch hydrolysis and base composition of DNA. 16 strains of the genus Prototheca do not produce extracellular amylolytic enzymes. The base composition of their DNA shows rather continuous values from 62% to 78% GC (guanine + cytosine). Their assignment to four species and their possible relationship with Chlorella protothecoides are discussed.


Turnover of lipids in Mycobacterium smegmatis CDC 46 and Mycobacterium phlei ATCC 354. The rates of breakdown and renewal of individual lipids in cultures of Mycobacterium smegmatis CDC 46 and Mycobacterium phlei ATCC 354 were investigated by means of a pulse labelling technique using palmitate-1-14C. The results indicated that in growing cultures of both strains phospholipids were broken down, and cardiolipin had a very rapid turnover. In chase experiments, almost 45% and 40% of the radioactivity of this component were lost respectively from M. smegmatis and M phlei during one generation time of the cell. The other two major components, phosphatidyl ethanolamine and phosphatidylinositol mannosides showed relatively low turnover. The loss of radioactivity from phosphatidylinositol mannosides was greater in M. phlei than in M. smegmatis but the loss of radioactivity from phosphatidyl ethanolamine was higher in M. smegmatis. The pattern of loss of radioactivity from lipids was almost the same in both strains, the difference being only in the extent of loss. The differences in the cellular localization of the phospholipids indicate their different roles within the cell. Results obtained with the glyceride fraction indicated a very rapid turnover of triglycerides in both strains.


Physiological and biochemical contributions to the taxonomy of the genus Chlorella. XI. DNA hybridization. 1. DNA homology was studied in 12 Chlorella species. The DNA of 88 strains was hybridized with 3H-labelled DNA from C. fusca var. vacuolata 211-8 b and from C. vulgaris 211-8m. The results indicate that the genus Chlorella is a heterogenous taxon which consists of several groups of species. 2. The "C. fusca group" comprises C. fusca var. vacuolata, C. fusca var. rubescens, and C. zofingiensis. Within this group, C. zofingiensis appears to be more closely related to C. fusca var. vacuolata than is C. fusca var. rubescens. C. fusca var. fusca does not belong to this group of taxa. 3. The "C. vulgaris group" consists of C. vulgaris, C. sorokiniana, and C. saccharophila. There are several strains which seem to assume a position intermediate between C. vulgaris and C. saccharophila. C. protothecoides is not related to this group of species. 4. Several groups of strains of C. sorokiniana with different base compositions (guanine + cytosine content) of their DNA appear to belong to the same taxon. 5. In addition to C. fusca var. fusca and C. protothecoides, also C. luteoviridis, C. minutissima, C. kessleri, and C. homosphaera seem to have so little relationship with the other species that their assignment to the genus Chlorella appears questionable.