Histological changes in thyroid of rat under the acute exposure of O-chloro-benzylidine malononitrile. This communication describes histological changes in the thyroid gland of rats under the acute stress of O-chlorobenzylidene malononitrile (CS) 10 mg/kg and 20 mg/kg). It has been observed that CS, when injected, causes histological changes in the thyroid of varying degrees, depending on the dose used.


A triple test for screening biological activity of prostacyclin analogues. 6,9-Thiaprostacyclin was used as a representative of prostacyclin analogues with the aim to design the most convenient procedure for evaluation of vasodilator, anti-platelet and other biological properties within this new group of potential anti-thrombotic agents.


Uptake of tolbutamide by islets of Langerhans and other tissues. 3-H-tolbutamide was distributed in a volume exceeding the space occupied by 14-C-sucrose in islets as well as in liver, kidney, muscle, and fat. In contrast to previous reports, the findings suggest that tolbutamide is not restricted to the extracellular space of islets.


[S-acylderivatives of thiosalicylamides with antifungal activity. I]. A series of S-acylderivatives of N-ethylthiosalicylamide (substances I leads to XXIII) was prepared and tested for in vitro antifungal activity. The substances, not previously reported, were prepared by the reaction of 2-mercapto-N-ethylbenzamide with suitable acylating agents. The fungistatic activity of the prepared products was tested in vitro against Candida albicans and Trichophyton mentagrophytes. The results given in Table I show that the S-acylderivatives of N-ethylthiosalicylamides have interesting antifungal activity. From examination of the results  (Tables I and II) some information on the structure-activity relationship was obtained. 2-Acetylmercapto-N-ethylbenzamide (I) and 2-propionylmercapto-N-ethylbenzamide (III) proved the most active of the compounds tested.


Effects of aging on the kidney.

Effects of aging on the kidney. A host of abnormalities of renal structure and function accompany advancing age. This presentation briefly surveys the renal anatomical and functional changes associated with senescence. Four areas of renal functional change have been selected for in-depth discussion: a) renal blood flow; b) glomerular filtration rate; c) renal sodium handling; and d) renal concentrating ability. The methodologic considerations including population selection which confound the assessment of the effects of aging on renal function are discussed. In addition, the functional changes associated with senescence are discussed in the context of longitudinal studies and studies utilizing appropriate patient cohorts, including potential kidney transplant donors. The clinical implications of senescent changes with regard to adjusting "normative" standards to fit the age of the patient are addressed. Furthermore, the implications of age-related renal functional alterations in predisposing the elderly patient to a number of fluid and electrolyte derangements are considered.--Epstein, M. Effects of aging on the kidney.


Sperm velocity and percentage of motility in 100 normospermic specimens analyzed by the multiple exposure photography (MEP) method. Specimens from 100 fertile or potentially fertile men with sperm counts above 20 million/ml were analyzed objectively by the new multiple exposure photography method (MEP). The following results (means and standard deviations) were obtained: sperm concentration, 66 million/ml +/- 47; percentage of motile spermatozoa, 45% +/- 18; speed of motile spermatozoa, 30.3 micrometer/second +/- 5.5; abnormal forms, 29% +/- 10.2; average motility index (product of sperm speed and fraction of motile spermatozoa), 14 +/- 7.5. Low correlations were found between sperm concentration, percentage of motility, and spermatozoal velocity. Moderate correlations were found between percentage of motility and spermatozoal velocity. Negative moderate correlations were found between abnormal forms and spermatozoal velocity as well as between abnormal forms and percentage of motility. The differences between some of our results and those obtained by others are discussed.


A simple inexpensive method for objective assessment of human sperm movement characteristics. A simple, objective method is described for obtaining a well-defined and diversified set of human sperm movement characteristics. These data are suitable for comprehensive statistical comparisons of sperm suspensions. One-second time-exposure photomicrographs are taken with dark-field illumination of slides prepared from semen, cervical mucus, or artificial media. The negatives are projected as a filmstrip and are analyzed on a specially designed console. The filmstrips can be taken and developed in 15 minutes without darkroom facilities, and a complete analysis of the film requires an additional 15 to 20 minutes. Fifty spermatozoa are examined to determine percentage motility, and movement characteristics are recorded for 15 spermatozoa. With these data the swimming trajectories of individual spermatozoa can be reconstructed retrospectively. The sperm suspension as a whole is described in terms of percentage motility, mean swimming speed, percentage of progressive spermatozoa, mean swimming speed of progressive spermatozoa, percentage of straight-swimming spermatozoa, percentage of rolling spermatozoa, and percentage of yawing spermatozoa.


Experimental studies on phototoxic and photoallergic reaction by the skin window technique. The differentiation of photoallergic contact dermatitis by 3,3', 4', 5-tetra-chlorosalicylanilide from phototoxic contact dermatitis by 8-MOP in guinea pigs was studied by skin window technique. Based upon the results, it may be concluded that the photoallergic reaction can be differentiated from phototoxic reaction by the appearing pattern of basophilic leukocytes.


The pathogenesis of lesions in Behçet's disease. Current concepts incriminate a cell-mediated mechanism in the pathogenesis of mucocutaneous lesions of Behçet's disease. However, such a mechanism alone fails to explain various aspects of hyperreactivity response which is a unique feature manifested with lesions similar to those occurring spontaneously in the disease. 19 patients with active Behçet's disease have been investigated for possible mechanisms other than cell mediated which may be involved in the process. Beside cell-mediated mechanisms there were indications that polymorphonuclear leucocytes and mast cells may play part in the process. Further study of these two aspects may open a new approach in the understanding and possibly in the treatment of this disease.


Evaluation of treatment of lepromatous leprosy patients in the Netherlands. The results of treatment of the group of leprosy patients at the lepromatous side of the leprosy spectrum registered at the Department of Dermatology of the University of Amsterdam in the years 1950-1976 were studied. The average duration of treatment to obtain bacteriologically negative skin biopsies in patients who were untreated at the time of registration, was 5 years. A substantial number of patients suffered a relapse; the main reasons for these relapses were discontinuation of treatment and DDS treatment in low dosage.


Synthesis of vitellogenin, an attractive model for investigating hormone-induced gene activation. The estrogen-induced synthesis of vitellogenin in the frog Xenopus and the chicken is an attractive system for investigating the molecular events leading to the activation of a specific gene. In this review article the events occurring at the level of the protein and mRNA in the cytoplasm are discussed. The available data show that the induction of vitellogenin synthesis is due to the accumulation of the corresponding vitellogenin mRNA in the cytoplasm. This suggests that transcriptional or posttranscriptional events in the nucleus are activated by the hormone. The few experiments investigating the processes in the nuclear compartment are reviewed.


Cell membrane, a target for steroid hormones. As yet there is little supportive evidence for the existence of a hormone-membrane interaction mechanism in somatic target cells, but there is no reason to exclude the possibility, either alone in certain cells or associated with the classical receptor system, since a hormone may well have more than one molecular target in a given cell. The present paper reviews studies of target cell membrane interactions with steroids, modified to prevent cell entry, in order to determine whether the interactions observed in oocytes are of wider significance.


Levamisole in Crohn's disease--a double-blind controlled trial. The efficacy of levamisole in patients with mildly active Crohn's disease was evaluated in a double-blind, placebo-controlled trial by means of clinical, biochemical, and immunological parameters. No immunosuppressive drugs were prescribed during the study. 11 patients received levamisole for 3 months (2.5 mg/kg body weight on 2 consecutive days/week) and 10 patients received placebo. Although the 2 patients groups were not very well comparable, the overall results of this study did not show obvious changes in clinical, biochemical, or immunological parameters in either group. No side effects of levamisole were observed during the study.


Daily elimination of fecal neutral sterols by humans. A study was made of the daily elimination of fetal neutral sterols in 22 healthy subjects and the average proportions of the main sterols were established. These were cholesterol (9.5%), its main bacterial transformation products: coprostanol (65%) and coprostanone (less than 6%), cholestanol (less than 2%), delta7-cholestenol (less than 2%) and phytosterols (18%). Epicoprostanol, which indicates a secondary mean of bacterial degradation of cholesterol has been searched for systematically. In general, traces only are present but in one case it amounts to more than 8% of the neutral sterols. The bulk of neutral sterols eliminated daily is very variable for different subjects and also for a same subject, although the concentration of fecal sterols is relatively constant (6.37 +/- 0.36 mg of sterols/g of feces). For a given subject under controlled diet conditions, differences in fecal elimination of sterols can essentially be ascribed to irregularity of digestive transit.


Effect of cimetidine on serum gastrin and gastric emptying in man. Paired studies of gastric emptying were performed in 10 healthy volunteers using a scintiscanning technique which provides information about the rate of emptying of both the liquid and solid components of a standard meal. Emptying rates of both components were not significantly different after oral administration of 400 mg cimetidine than after placebo. The increase in serum gastrin concentration produced by the meal was significantly greater after cimetidine than after placebo but there was no relationship between individual cimetidine-placebo differences in gastrin response and the corresponding cimetidine-placebo differences in rates of emptying. A positive correlation was demonstrated between the gastric emptying rate of the liquid component and the 1 h cimetidine absorption, indicating that normal individual variation in gastric emptying is probably responsible for much of the normal individual variation in the rate of cimetidine absorption.


Stimulation of gastrin release and gastric secretion: effect of bombesin and a nonapeptide in fistula dogs with and without fundic vagotomy. Bombesin and a synthetic bombesin nonapeptide were studied by intravenous infusion at a dose of 0.5 microgram.kg-1.h-1 for 4 h in 7 dogs with esophagostomy and gastric fistula. In 3 of the dogs who had highly selective (fundic) vagotomy, mean integrated gastrin output over 4 h was double that in the 4 dogs with vagi intact during both nonapeptide (1,554 vs. 700 pg.ml-1.4 h-1) and bombesin infusion (2,442 vs. 1,440 pg.ml-1.4 h-1). Peak concentrations of serum gastrin reached during bombesin (490 +/- 100 vs. 320 +/- 90) were higher than those during nonapeptide infusion (270 +/- 40 vs. 160 +/- 28 pg/ml) in the vagotomized and intact dogs, respectively. The difference between vagotomized and vagally intact dogs suggests that the fundic vagotomy removed an inhibitor of gastrin release from the innervated antrum. Despite these differences in gastrin release, gastric acid output with the two peptides was the same (49--52 mEq/4 h) whether the fundus was denervated or innervated. This suggests that bombesin may stimulate gastric acid secretion by the release of an additional secretagogue which is not measured by the gastrin assay. Neither of the two inhibitors of gastrin release--antral acidification to pH 1.4 or less or atropine (100 microgram/kg)-- inhibited gastrin release by bombesin, even though the atropine reduced acid output by 80%. Bombesin is a potent gastric stimulus whose action is only partly explained by the measured gastrin release.


Possible role of endogenous prostaglandins in glucagon secretion by isolated guinea-pig islets. Previous studies have demonstrated that prostaglandins stimulate glucagon secretion in vitro and in vivo. The present work was aimed at investigating the influence of two inhibitors of prostaglandin synthesis, isopropyl-2 nicotinoyl-3 indole (L8027) and indomethacin, on basal and arginine- or noradrenaline-stimulated glucagon release from isolated guinea-pig islets incubated in the absence of glucose. L8027 (10(-4) and 10(-5) mol/l) did not alter basal glucagon release, blocked almost completely the glucagon response to arginine (10(-2) mol/l), had no effect on the glucagon release induced by noradrenaline (10(-4) mol/l), but reduced the stimulatory effect of a lower concentration of noradrenaline (5.10(-7) mol/l). The kinetic study of this inhibitory effect demonstrated that (1) it necessitates preincubation of the islets with L8027 for 30 minutes before the addition of arginine, (2) after a short preincubation period (30 minutes) in the presence of L8027, removal of the inhibitor at the time of arginine stimulation resulted in enhanced glucagon response, (3) on the contrary, after a prolonged incubation period (75 min) with arginine and L8027, the inhibitory effect remained transiently detectable after removal of L8027. Indomethacin similarly blocked arginine- and noradrenaline-induced glucagon secretion. These results suggest that an intra-insular synthesis of prostaglandins is involved in the A cell response to arginine and noradrenaline.


The influence of amyloid deposits on the islet volume in maturity onset diabetes mellitus. Pancreatic islet volumes of patients with and without maturity onset diabetes mellitus were estimated. The islet volume of the diabetic patients was 1.01 +/- 0.12 cm3 (SEM) and that of the non-diabetic patients 1.60 +/- 0.16 cm3 with considerable overlap between the two groups. Islet amyloidosis was found in all the diabetic and in 9 of the 15 non-diabetic patients. When the amyloid deposits were excluded, the islet volume of the diabetic patients was 0.89 +/- 0.10 cm3, while that of the non-diabetic patients was unchanged, 1.60 +/- 0.16 cm3. There was still some overlapping. Since amyloid deposits seem to destroy the B cell membranes, it was postulated that a comparison of the volumes of islets completely free of amyloid might give a more true picture of the quantitative islet alterations in maturity onset diabetes. It was found that this islet volume of the diabetics was only 0.41 +/- 0.05 cm3 and that of the non-diabetic patients 1.58 +/- 0.16 cm3. These values correspond better to the altered insulin secretion in maturity-onset diabetes mellitus.


Insulin deficient diabetes. Contrasts with other endocrine deficiencies. Comparisons are made between the incidence, prognosis and treatment of juvenile-onset diabetes and other endocrinopathies in the young. 548 patients with insulin deficient diabetes diagnosed before 20 years of age have been reviewed. Excess mortality, especially at 35--40 years of age was found. Profiles of blood glucose and serum insulin have been studied and compared to those of normal subjects. The variation of insulin absorption and effect of insulin antibodies on the free insulin levels achieved after exogenous insulin injections have been demonstrated. The common occurrence of nocturnal subclinical hypoglycaemia following intermediate or long-acting insulin was often found to be the cause of poor diabetic control. Five out of 33 patients with 'difficult' diabetes had an unexplained resistance to high levels of free-insulin. The value of self-monitoring and HbAl measurements in the improvement of diabetic control and possibly life expectation is reviewed. The incidence of thyroid disease was found to be increased in 1779 insulin deficient diabetics of all ages and persistence of islet-cell antibodies suggests that the diabetes may be due to autoimmunity in some of these patients.


[Effectiveness of two Ca-antagonistic drugs on angina pectoris: nifedipine and verapamil versus nitroderivative and beta-blocking drugs (author's transl)]. The Authors, using cycle ergometer tests, assess the effectiveness of two Ca-antagonistic drugs, nifedipine and verapamil, administered full strength to patients suffering from typical angina from effort. The true effectiveness of these drugs was tested by comparing the results with those obtained with placebo, isosorbide dinitrate and propranolol. The effect of nifedipine proved statistically significant (p less than 0.01) with respect to the parameters: double product, time of insurgence of angor and time of appearance of electrocardiographic anomalies. Verapamil proved slightly significant (p less than 0.05) only with regard to time of appearance of electrocardiographic anomalies. There follows a brief discussion of the differences between these results. Isosorbide dinitrate and propranolol increase tolerance to effort with respect to placebo treatment. These results are statistically significant on all parameters used.


[Effect of metoprolol on the exercise tolerance in patients with stable angina pectoris (author's transl)]. The effect on the exercise tolerance induced by a new selective beta1-receptor antagonist, metoprolol, given by a single administration per os, has been evaluated in 16 patients with stable angina pectoris. The study was a double blind cross-over between metoprolol 100 mg and placebo. Patients were hospitalized for the time required for the performance of the study; every cardioactive drug, except for trinitrine, was stopped 72 hours prior to the study period. Exercise testing was performed two hours after the intake of the drug, in a sitting position on a bicycle ergometer; the workload was increased by 25 Watts increments every two minutes. Compared to placebo, the total workload performed and duration of exercise increased after metoprolol (P less than 0.001), heart rate decreased (P less than 0.001) both in the resting state and during exercise test, systolic blood pressure didn't change in the resting state but decreased during exercise test (P less than 0.01), pressure-rate product, an useful index of changes in myocardial oxygen consumption decreased both in the resting state (P less than 0.001) and during exercise test (P less than 0.001). It is concluded that metoprolol induces a significant increase in exercise tolerance. This is due to a marked reduction in oxygen myocardial consumption resulting by both decreased heart rate and systolic pressure.


[Pregnancy and delivery after renal transplantation (author's transl)]. A case report is given on a 21-years-old woman who became pregnant 18 months after renal transplantation. In spite of relatively unfavourable conditions (match 0,25; hypertension) renal function showed a good adaptation throughout the gestation. In the 38. week of pregnancy the woman was successfully delivered of a 2440 g healthy female child by vacuum extraction. There were no effects of the immunsuppressive drugs on the newborn. It is the third case of a successful delivery after renal transplantation in the GDR.


Gastrointestinal radiography with glucagon. This report summarizes the results of nine diagnostic radiographic studies done double blind crossover comparing glucagon to placebo and to anticholinergic drugs in volunteers. In seven studies the subjects were administered drug intramuscularly and in two studies intravenously. There were five diagnostic studies of the upper gastrointestinal tract, one for esophageal varices and three of the colon. The results indicate that glucagon can be given intramuscularly and intravenously. When given intravenously it has a rapid onset and predictable length of action depending on the dose given. Reports of side effects were few consisting primarily of nausea and or vomiting. These results indicate that glucagon is the drug of choice for hypotonic diagnostic examinations.


Effect of intravenous diazepam on human lower oesophageal sphincter pressure under controlled double blind crossover conditions. The effect of diazepam on the lower oesophageal sphincter (LOS) pressure is controversial. Therefore, a double-blind crossover study was performed on 18 healthy volunteers to determine the sphincter response to intravenous diazepam--70, 140, 280 microgram/kg, which correspond to a total dose of 5, 10, and 20 mg. respectively. After the 5 and 10 mg dose no signficant effect on LOS pressure could be observed when compared with placebo. After the 20 mg dose a significant rise in pressures (deltaPLOS) was recorded for 40 minutes with a maximum deltaPLOS of + 16.2 +/- 6.6 (mean +/- SEM) mmHg after 50 minutes (P less than 0.01) (46 +/- 1.3% increase above the basal pressure). It is concluded that diazepam does not affect lower oesophageal sphincter competence and therefore does not increase the risk of regurgitation and pulmonary aspiration in premedicated patients.


Comparison of two doses of cimetidine and placebo in the treatment of duodenal ulcer: a multicentre trial. Endoscopy, clinical assessment, and laboratory studies were used to compare, in a double-blind multicentre trial, the effects on patients with duodenal ulceration of treatment for four weeks by either placebo or 1 g/day cimetidine, or 2 g/day cimetidine. Ulcer healing occurred in 28% of patients on placebo, 61% of patients on 1 g cimetidine daily, and 70% of patients on 2 g cimetidine daily. Thus cimetidine conferred an advantage over placebo, but the effects of the two doses of cimetidine were not shown to be different. Symptomatic improvement in patients given cimetidine was usually marked and occurred early. Patients were required to report all symptoms, but the only symptom which might have been caused by cimetidine was headache in 5% of patients. Biochemical studies showed significant (though slight) rises in serum uric acid, and serum creatinine but no significant changes occurred in the serum levels of liver enzymes. This study confirms that 1 g is a suitable daily dose of cimetidine for the treatment of duodenal ulceration.


[Iron deficiency in infection].

[Iron deficiency in infection]. As a rule bacterial infection is followed by acute serum iron reduction; impaired inflow of iron from storage sites into the transport pool represents the main cause. As a consequence of this sort of iron redistribution iron becomes short for red cell production; this is one cause for the development of anemia. The biological significance of hyposideremia may be presumed from the bacteriostatic potential of iron free transferrin, preventing adequate iron acquisition by multiplying microorganisms. Preliminary animal experiments support this concept and suggest that it might also be applied to true iron deficiency.


Evaluation of semen quality following kallikrein treatment. 30 asthenozoospermic subfertile men were treated parenterally for 30 months with kallikrein. 20 men showed a significant increase in sperm motility, 66% (responders), while the other 10 men, 34% (non-responders), did not respond to this treatment. A striking decrease in sperm concentration (by 32%) was found in most patients. No latent period was found following kallikrein treatment. Thus, semen quality did not differ if examined immediately following the cessation of treatment or a few weeks afterwards. A poor pregnancy rate was recorded, i.e. 20% among the responders and 10% among the non-responders. In conclusion, kallikrein is a useful drug in the treatment of asthenozoospermia only when the sperm concentration is not decreased below the optimal value.


Effect of flufenamic acid on uterine contractions and plasma levels of 15-keto-13,14-dihydroprostaglandin F2alpha in preterm labor. Flufenamic acid (FA), an inhibitor of the synthesis and action of prostaglandins, was administered to 18 women with preterm labor during the 28th to 36th week of gestation. In 15 patients delivery postponed, the mean admission/delivery interval being 21.5 days. 2 patients with cervical dilatation of 4 cm delivered within 48 h despite medication. The peripheral plasma levels of 15-keto-13,14-dihydroprostaglandin F2alpha (KH2 PG2alpha) was high on admission (216 +/- i4 pg/ml, mean +/- SEM), declined by 50% within 2 h of instituting treatment and remained near the normal level seen in 13 controlled women after the 24th hour.


Major proteins of the Escherichia coli outer cell envelope membrane. Sequence of the cyanogen bromide fragments of protein I from Escherichia coli B/r. The sequence of the cyanogen bromide fragments of one of the major outer membrane proteins of E. coli B/r has been established with the aim of elucidating the primary structure of this protein. Separation of all fragments on one molecular sieve column was achieved upon citraconylation of these fragments. Overlapping peptides were obtained by digestion of the protein, or a cyanogen bromide fragment arising from incomplete cleavage, with trypsin or Staphylococcus aureus protease.


Psychiatric units in general hospitals: 1979. Despite the network of community mental health centers, the general hospital has become the focal point for the delivery of mental health care in the U.S. The author presents an overview of the psychiatric unit in the general hospital, including its history, structure and function, and its relationship to the hospital itself and to the continuum of mental health services in the community. The units' goals are not clearly defined but appear to be crisis intervention, acute treatment, correction of decompensation, prevention of chronicity, and speedy return of the patient to the community; there is little attempt to serve chronic patients. Paradoxically, the psychiatric unit also does not serve the hospital it is part of, as it rarely accepts patients from medical-surgical wards. The author summarizes evaluation studies related to general-hospital psychiatric units and recommends, among other points, truly evaluating the effects of short-term treatment and eliminating the current competition for the shortest stay.


Governance: trustees are closing the gap between hospitals and consumers. Many trustees today believe that they are in the best position to cement the relationship between their hospital and its service population.


Human relations: will humanism and public service survive? Bureaucratization and specialization threaten to undermine personalized care. Will humanism and public service be sacrificed for the ethos of business?


Labor relations: union activity increases among professionals. Hospital labor relations managers need a higher level of skills today to deal with increasingly complex labor-management negotiations.


Marketing: fast becoming a necessary tool for hospital administrators. Many hospitals need more business, and marketing techniques have been suggested in the literature as one way to attract it.


Medical staff: physicians seek new responses to mounting pressures. In responding to pressures to contain cost and improve the quality of care, medical staff are becoming increasingly involved in policymaking and managerial decisions.


Multi-institutional arrangements: hospitals' move toward systems gains momentum. The health field continues to restructure itself through multi-institutional and sharing systems that aim at ensuring improved and cost-effective services.


Quality assurance: debate persists on goals, impact, and methods of evaluating care. Experts asked whether 'quality assurance' improves care and utilization, cuts costs, changes behavior, needs new formats, meets new uses, or poses conflicting values.


Management engineering: branching out to many areas of the hospital. Much emphasis is given to the quantitative techniques of scheduling and forecasting to reduce costs and improve efficiency.


Regulations: hospitals' battle against excess legislation continues. Recent experiences have given the health care industry different approaches it can use to deal more effectively with government regulations.


Risk management: idea and effort widely supported, but appropriate methods debated. Debate continues on use of self-insurance, incident reports, closed claims studies, and medical and peer review records.


Cost containment: are hospitals taking a bum rap? The private market has been distorted by government regulations and programs; more reliance on market incentives rather than on regulation is needed.


Education and training: fulfilling the demands of the future. Hospital-based educators must establish boundaries for their profession and find new ways to manage educational programs within the institution.


Energy: hospitals apply total management approach. Energy-conscious hospitals are gearing up to conserve in anticipation of higher prices and diminishing supplies.


Emergency services: growing discipline comes of age. The first explosion of growth in EMS systems is followed by assessment, information-gathering, and building on programs already in place.


Financial management: hospitals need sophisticated techniques. For financial survival, hospitals must use a wide variety of methods, ranging from operational audits and profitability analyses to sharing and improved collection efforts.


Financing and accrediting hospices.

Financing and accrediting hospices. Attention by third parties to critical licensure and reimbursement issues is the appropriate response to growing public support of hospice care.


Demonstration of replication patterns corresponding to G- and R-type banding of chromosomes after partial synchronization of cell cultures with BrdU or dT surplus. A standard protocol is reported for the highly efficient demonstration of replication patterns corresponding to R-type and G-type banding.


The testis as a secretory organ for H-Y antigen. After cultivation of dissociated rat testicular tissues, H-Y antigen is detectable in the medium; this is not the case if nongonadal male tissues are incubated. Release of H-Y antigen by testis cells is inhibited by the addition of cycloheximide. All tissues still type H-Y positive after culture. It is assumed that the testis actively secretes H-Y antigen. This assumption is supported by the finding that the amount of H-Y antigen in the epididymal fluid increases with the age of the animals.


Acid alpha-glucosidase in Malaysians. Population studies and the occurrence of a new variant. Acid alpha-glucosidase from the placenta was electrophoretically surveyed in a total of 633 Malaysians, 236 of Malay, 261 of Chinese and 136 of Indian ancestries. A new variant, alpha-glucosidase 3-1 was observed in 1 Malay and 3 Indians. A polymorphism for this enzyme was observed among Indians, but in Chinese and Malays variants are rare. Phenotype 2-1 was observed once in a Chinese and once in a Malay.


Do natural killer cells engage in regulated reactions against self to ensure homeostasis? Host reactivities not requiring immunization in the mouse, especially natural resistance of irradiated animals to accept grafts of normal or malignant hemopoietic cells, were compared with NK activity against the YAC-1 lymphoma. The effects of several independent variables known to influence natural resistance in vivo had a similar effect on the NK system. Figure 12 lists an impressive array of shared properties and positive correlations. In contrast, the distinctions were few and minor. Many of the positive correlations were of particular significance since the experimental variables either have opposing or no effects on conventional induced immunity. The multiplicity and pervasiveness of these correlations suggest that the cellular mechanisms underlying natural reactivities are similar or common. Cytotoxic effectors mediating natural resistance to normal cells, tumors, and cells infected with intracellular pathogens may be distinct in terms of target selectivity, yet belong to a single cell lineage subject to common regulatory influences for differentiation and function. Regulation of reactivity via suppressor cells was studied in the NK system only. The spleens of mice selected for low levels of NK activity (resulting from young age, irradiation, and treatment with the macrophage-active agents l-carrageenan or hydrocortisone acetate) contained cells capable of inhibiting the lytic function of NK effectors taken from untreated adult donors. All the suppressor cells studied were thymus-independent, as judged by their occurrence in spleens of genetically athymic mice; the suppressive function was resistant to 2000 rads of gamma-rays administered in vitro and was not restricted by the major histocompatibility complex, without exception. However, two major classes of suppressors were identified: (a) macrophagelike cells inducible by l-carrageenan or hydrocortisone acetate, and (b) nonadherent cells found in spleens of untreated infants and of irradiated adult mice. It is proposed that the suppression of NK cytolysis demonstrated in vitro was a manifestation of regulatory mechanisms modulating the level of NK activity in vivo. Macrophagelike cells that are induced, activated, or inactivated by bacteria, viruses, hormones, and other agents may act as regulators of differentiation, maturation, and function of cells belonging to the NK lineage. Nonadherent cells could be either a distinct class of suppressors or immature NK cells capable of binding but not lysing target cells. In the latter case, regulation would be achieved via competitive binding of targets by pre-NK cells presumably in dynamic equilibrium with functional (i.e. matured) NK effectors.


Effects of cyclophosphamide on the in vivo response of outbred athymic (nude) mice to a thymus-independent antigen (DNP-AGG-Ficoll). Both nude mice (nu/nu) and their heterozygous littermates (nu/+) were injected with a single IP dose of 300 mg cyclophosphamide (CY)/kg. CY is a known immunosuppressive agent, which affects primarily B lymphocytes. Immunization with the thymus independent antigen DNP-AGG59-Ficoll after CY treatment disclosed that restoration of the primary direct PFC response occurred more rapidly in nude mice than in nu/+ mice. However in these same experiments, the primary indirect PFC response, recovered earlier in nu/+ mice than in nude mice. After CY treatment, secondary indirect PFC responses were delayed in both nude and nu/+ mice, but the greatest effect was seen in nude mice. The data suggest that the presence of T cells has little if any influence on the recovery capacity of those B cells which are destined to become direct PFC. However the recovery of B cells which are destined to produce indirect PFC responses is facilitated by the presence of T cells.


The effect of the parenteral administration of a rabbit anti-(mouse)-IgD serum on the immune response of mice to sheep erythrocytes. Experiments have been carried out to find out if the administration of an anti-IgD serum to mice interferes in anyway with their immune response to sheep red blood cells. This was done to test a hypothesis that a biological role for IgD might be as a critical cellular receptor for antigen. Our results show that the injection of anti-IgD two days before antigen results in suppression of primary responses and priming (the antigen dependent generation of memory cells) but has no suppressive effect on a secondary response. On this indirect evidence we conclude that it is likely that IgD is present on antigen-sensitive percursor cells but not on memory cells.


Allelic exclusion of M1 (IgM) allotype on the surface of chicken B cells. The membrane expression of M1 (IgM) and G1 (IgG) allotype markers on peripheral blood lymphocytes was examined by immunofluorescence. In homozygous chickens 15% lymphocytes stained with either anti-M1 or with polyspecific rabbit anti-Ig serum, suggesting that M1 is expressed on the surface of probably all B lymphocytes. In heterozygous M1a/M1b individuals antisera against either allelic antigens reacted with 50% of the total surface Ig positive cells as evidence for the allelic exclusion of surface M1 expression. Donor allotype synthesis was determined from serum allotype levels in congenic cyclophosphamide-treated recipients of lymphoid cells. Treatment of cells with anti-M1 serum prior to transfer inhibited both M1 and G1 allotype synthesis. The treatment of heterozygous (M1a/M1b, G1a/G1e) cells with anti-M1b serum inhibited specifically the synthesis of M1b and G1e allotypes controlled by linked genes from the same parental chromosome. Thus, B cells which had been the targets for anti-M1 antibody mediated suppression are also subject to allelic exclusion.


Lipid chemotaxins isolated from culture filtrates of Escherichia coli and from oxidized lipids. Lipid extracts of sterile culture filtrates of Escherichia coli were shown to contain approximately 75% of the chemotactic activity for human polymorphonuclear leukocytes and rabbit alveolar macrophages. Fractionation and purification of these lipids revealed the presence of many unknown lipids of widely different properties, but all were anionic and at very low concentrations, chemotactic. The only one of active molecules that could be identified was an unsaturated ultraviolet-absorbing hydroxy fatty acid, which, following catalytic reduction with hydrogen, was found to be hydroxyeicosanoic acid. This fatty acid's chromatographic behavior was very similar to that of 12-hydroxy-5,8,10,14-eicosatetraenoic acid (HETE), which is a potent chemotaxin for polymorphonuclear leukocytes and macrophages. Unknown chemotaxins could be generated by the oxidation of known unsaturated lipids. Prostaglandins A2 and E2 produced potent chemotaxins upon aerobic oxidation. Malonaldehyde, a peroxidation product of unsaturated lipids, when reacted with phosphatidylethanolamine in aerobic conditions, also produced strong chemotactic agents. The chemotactic activity of these products could be destroyed by catalytic reduction with hydrogen and by methylation with dry methanolic HCl. These data indicate that the nonenzymatic oxidation of unsaturated lipids generates some products that are potent chemotaxins for mammalian inflammatory cells.


Complement-dependent hemodynamic and hematologic changes in the rabbit. The intravenous injection of the anticomplementary protein from cobra venom, cobra factor (CoF),2 induces decreases in mean arterial blood pressure and circulating platelets in rabbits. The changes are rapidly reversed. Both changes require the presence of C3 and occur in rabbits genetically deficient in the sixth component of complement. The hypotensive effects of CoF were blocked by the histamine H2-receptor antagonist burimamide. An acute C3-dependent change in blood pressure and circulating platelets also was demonstrated following the intravenous injection of S. marcescens endotoxin. However, abrogation of these acute changes by C3 depletion did not alter the extent of a second, prolonged fall in blood pressure and platelets induced by S. marcescens endotoxin occurring after 60--90 min. C3 depletion also did not alter the lethal effects of the S. marcescens endotoxin.


Treatment of rheumatoid and degenerative diseases with copper complexes: a review with emphasis on copper-salicylate. This review presents a historical account of the treatment of rheumatoid and other degenerative diseases with copper complexes. Clinical data obtained from 1940 to 1971 are provided for about 1,500 patients with rheumatoid arthritis (acute or chronic), rheumatic fever, ankylosing spondylitis, staphlococcal spondylitis, gonococcal arthritis, chronic gouty arthritis, polyarticular synovitis, coxitis, disseminated spondylitis, arthritis with psoriasis, Reiter's syndrome, lupus erythematosus, sarcoidosis, arthrosis deformans, erythema nodosum, sciatica (with and without lumbar involvement), cervical spine-shoulder syndrome or lumbar spine syndrome. The drugs used in these studies were Dicuprene, Alcuprin, Cuprimyl, and Permalon, a copper-salicylate preparation. A detailed presentation of toxicities associated with the use of these copper complexes is included.


Non-conventional treatment of hepatic failures. It is our intention to present a short review of various approaches to the non-conventional treatment of hepatic failures of the fulminant type. Our review is directed to the scientist, technologist, and clinician with a budding interest in the hepatic assist area. We shall discuss parabiosis, liver transplants, and various extracorporeal devices including hemoperfusion, hemodialysis, and enzymic detoxification systems. We feel that the present technological approaches to the treatment of hepatic failure are very primitive at this stage. Some of the recent advances are very encouraging, and it is our opinion that these approaches show great promise in the long term.


Some operations research applications to problems of health care systems (a survey). In this paper, a brief survey of operations research applications to problems of health care systems is made. It shows the possibilities which exist for improving the design and operations of hospitals and other health care systems through the use of operations research approaches. The different applications have been divided into groups, but it should be noted that there is a great deal of overlapping among them.


Computer-based testing system for experiments in visual perception. This paper discusses the increasing scope for the application of computing systems to the investigation of problems in the field of visual perception. A simple language for the generation of various perceptual testing schemes is described and some examples of possible applications are given.


Computer-based interpretation of clinical laboratory tests. A system has been developed for the interpretation of laboratory findings which can aid in medical diagnosis as well as monitoring medical treatment. Tools are provided for defining, updating and verifying rules for the interpretation of laboratory tests in a computer-acceptable form. The present system uses a set of arithmetic logic rules which allows comprehensive evaluation of laboratory data as well as bringing into focus fluctuating and borderline situations and inconclusive data. Application of the system for over a year has shown that it is easy to use by laboratory staff with no knowledge of programming. Moreover, it has forced greater comprehensiveness, objectivity, consistency and speed in the interpretation of laboratory results than is generally achieved by standard practice.


Peak-detection algorithm for EEG analysis. A peak-detection method is described for computer analysis of the the electroencephalogramme (EEG). The technique consists of measuring the amplitude and time interval between successive maxima (peaks) and minima (troughs) in the signal. A critical feature of the peak-detection algorithm is the inclusion of an amplitude threshold criterion which eliminates the registration of low-voltage activity riding on EEG waves. The peak-detection procedure permits the formulation of a variety of intra-band and inter-band EEG statistics which can be useful in on-line computer applications. The peak-detection algorithm has been successfully applied to a number of normal and clinical EEG recordings. Although no computer procedure for EEG analysis has yet been universally adopted, the peak-detection algorithm reported in this paper presents a standardised approach which can be used between EEG clinics.


Development of a computerised cancer data management system at the Mayo Clinic. A multidisciplinary team was assembled to design a cancer data management system that would meet the storage, retrieval and analysis NEEDS OF Mayo's clinical trials of anti-tumour drugs. To fulfill these requirements, a computerised data entry and retrieval system was developed, the primary patient health record used by the oncologists was redesigned and the clerical procedures and work flow within the Cancer Center Statistics Office were reorganised. The end result of this project is a system that has: (1) enabled Mayo to meet its reporting requirements as a Comprehensive Cancer Center; (2) provided the statisticians and the physicians with the capacity to perform more detailed and accurate analyses of clinical trials; (3) reduced the clerical effort needed for preparing reports and analyses; (4) provided the potential for expansion to meet the growing requirements of the future and (5) attained that often elusive goal of computer systems--user satisfaction.


K562--a human erythroleukemic cell line. We have studied the surface membrane properties of the human leukemic cell line K562 which previously has been reported to represent an early stage of granulocyte maturation. The surface glycoprotein pattern of the K562 cells obtained after galactose oxidase-NaB[3H]4 labelling and slab gel electrophoresis shows striking similarities with that of normal erythrocytes but is completely different from the patterns of normal and malignant cells of various stages of the myeloblast to granulocyte differentiation. Moreover, the K562 cell expressed the major red cell sialoglycoprotein, glycophorin, on its surface as shown by immunofluorescence and by immunoprecipitation from labelled membrane preparations. As glycophorin is exclusively found on erythroid cells in human bone marrow we conclude that the K562 is a human erythroleukemic line.


Pirbuterol in the treatment of bronchial asthma. Pirbuterol, a new anti-asthmatic compound, was tested in 21 male outpatients with reversible bronchospastic disease during 12 months in different dosis regimens in order to determine the extent and duration of bronchodilator activity, the optimum multiple dose-schedule, the incidence, type and degree of side effects, and the cardiovascular as well as hematologic, renal and hepatic function after oral administration. All patients were checked by monthly pulmonary function studies, E.C.G. and extensive laboratory tests. Aso slit lamp examinations were performed. Pirbuterol is a well-tolerated bronchodilator which appears to be of clinical importance in the long-term treatment of bronchospastic disease.


A comparative study on the clinical effects of oxazepam and diazepam: Relationship between plasma level and effect. The clinical effects of oxazepam and diazepam as oral premedicants were tested in a double-blind study of 60 children and 50 adults. The gas chromatographically measured concentrations of the active unconjugated forms of oxazepam and diazepam in the plasma were correlated to their clinical effects, as assessed both subjectively and objectively (sleep, sedation, apprehension, excitement, dizziness, emetic effect, headache, increase or decrease in systolic blood pressure, increase in pulse rate, venepuncture). No significant difference in the effects of these two benzodiazepine derivatives were observed, nor was there any obvious relationship between the plasma concentration and clinical effect.


Hypocholesterolemic effect of ascorbic acid in maturity-onset diabetes mellitus. A significantly lower vitamin C concentration has been found in the blood and particularly in the leukocytes of hypercholesterolemic diabetic patients than of healthy blood donors. Ascorbic acid administered in a dose of 500 mg per day for 12 months to metabolically stabilized hypercholesterolemic subjects with maturity-onset diabetes mellitus (diabetic diet without insulin or diabetic drugs) brought about a striking decline of cholesterolemia and a moderate decline of triglyceridemia. The serum lipid level in the control group given placebo remained unaltered. A daily administration of 500 mg of ascorbic acid for six months failed to affect the fasting level of serum immunoreactive insulin. It is assumed that the long-term administration of ascorbic acid to maturity-onset diabetics removed the tissue ascorbate deficiency and improved the liver ability to compensate the increased endogenous synthesis of cholesterol by its enhanced transformation to bile acids.


Disappearance of oxytocin-induced uterine tiredness by treatment with fructose-1,6-diphosphate. Experimental evidence. Fructose-1,6-diphosphate (FDP) is able to abolish oxytocin induced spastic inertia in rat uterus. The clinical use of FDP is suggested by observation carried out on 30 deliveries with oxytocin induction. The FDP-treated patients (5 g of FDP in 50 ml of water by intravenous infusion) showed a statistically significant decrease of time elapsed between the beginning of uterine inertia and the recovery of uterine contractions (176 +/- 25.4 min) compared to controls (562 +/- 32.5 min).


A technique for recording abutments and tissue relationships and inter-arch relations in one session. A modified technique for recording tissue and abutment relationships in one session has been presented. Using the technique chair-tite and discomfort for the patient are significantly reduced.


The overdenture: a concept.

The overdenture: a concept. Teeth that are unsuitable as abutments for a fixed bridge or removable partial denture may be retained as roots under an overdenture. While these roots remain, the vertical height of the bony ridge is preserved. In addition, the roots give support to the denture and reduce stress on the alveolar ridge.


Marginal leakage in amalgam restorations and its prevention. Marginal leakage is the penetration of fluids, bacteria and ions into the space existing between all restorative materials and cavity walls. It can cause pulp irritation, colour changes in the tooth and secondary caries, and may result in failure of the restoration. The margins of a fresh amalgam restoration invariably leak. As time passes there is a decrease in marginal leakage related to the production of corrosion products and the spreading of certain materials into the space between the tooth and the restoration. Correct cavity margin preparation and proper condensation of the restorative material minimize marginal leakage round fresh amalgam restorations. Varnish coated on the cavity walls is most important and an indispensable method for preventing marginal leakage.


Surgical demand scheduling: a review.

Surgical demand scheduling: a review. This article reviews the literature on scheduling of patient demand for surgery and outlines an approach to improving overall performance of hospital surgical suites. Reported scheduling systems are categorized into those that schedule patients in advance of the surgical date and those that schedule available patients on the day of surgery. Approaches to estimating surgical procedure times are also reviewed, and the article concludes with a discussion of the failure to implement the majority of reported scheduling schemes.


Pancreatic glucagon cells contain endorphin-like immunoreactivity. Semithin and thin serial sections of rat pancreas were investigated by immunocytochemical techniques using antisera against glucagon and endorphin. The immunocytochemical staining methods revealed that in the rat endocrine pancreas endorphin-like immunoreactivity is contained in glucagon (A-) cells. All secretion granules of glucagon cells that could be identified in adjacent thin sections exhibited glucagon as well as endorphin immunoreactivity. Since glucagon cells are devoid of ACTH-like immunoreactivity it is concluded that the biosynthetic pathway of pancreatic endorphin is not the same as in the pituitary.


The scope of a laboratory animal program needed at a veterinary school. The discipline of laboratory animal medicine is one of the most rapidly expanding specialties within the veterinary profession. Veterinary schools should fully accept the responsibility for introductory instruction in laboratory animal medicine in the professional curriculum. Such instruction should articulate the varied opportunities that exist for the laboratory animal veterinarian within the biomedical research community, and provide an overview of the normal biological characteristics and pathologic conditions of the common laboratory animal species. In addition, the opportunity should exist within the veterinary school for graduate and undergraduate students utilizing experimental animals to receive a comprehensive introduction to laboratory animal biology, care, and management. Instructional responsibility for such courses should be accepted by faculty veterinarians with advanced training in laboratory animal medicine. Veterinarians with advanced training in this specialty are uniquely qualified to make substantial contributions to biomedical research by promoting the health and welfare of the research animal.


New cephalosporins with 7-acyl groups derived from beta-ketoacids. I. 7-(beta-Ketoacylamino)cephalosporins. The synthesis and antimicrobial profile of a series of 7-(beta-ketoacylamino)cephalosporins (1) bearing an acetoxymethyl or a heterocyclichiomethyl group at the 3-position are described. Of this series, 3-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-7-(3-oxobutyrylamino)ceph-3-em-4-carboxylic acid (11) showed moderate antibacterial activities in in vitro and in vivo tests.


New cephalosporins with 7-acyl groups derived from beta-ketoacids. II. Further modifications of 7-(3-oxobutyrylamino)-cephalosporins. New cephalosporins modified in the acyl part of 7-(3'-oxobutyrylamino)cephalosporins (1), which have been described in the preceding paper, were synthesized by thiolation at the 2'- or the 4'-position, or by transforming the 3'-oxo group into a 3'-imino group. The most active compound in vitro was 3-[[(1-methyl-1H-tetrazol-5-yl)thio]methyl]-7-(4-methylthio-3-oxobutyrylamino)ceph-3-em-4-carboxylic acid (7c), which showed superior in vitro activity against Gram-positive and Gram-negative bacteria compared to the parent cephalosporin (1b) with the same 3-substituent. The ED50 value for 7c, however, was essentially equal to that of 1b in mice infected with Escherichia coli O-111.


A new cephalosporin. SCE-963: 7-[2-(2-aminothiazol-4-yl)-acetamido]-3-[[[1-(2-dimethylaminoethyl)-1h-tetrazol-5-yl]-thio]methyl]ceph-3-em-4-carboxylic acid. Chemistry and structure-activity relationships. The synthesis and the in vitro and in vivo antimicrobial activities of a series of 7-[2-(2-aminothiazol-4-yl)acetamido]cephalosporins (1) having varied 3-substituents, such as methyl, hydroxymethyl, acetoxymethyl, pyridiniomethyl and heterocyclicthiomethyls, are described. The derivatives having five membered heterocyclicthiomethyls exhibited strong inhibitory activities against Gram-negative organisms including some strains of Escherichia coli and Proteus morganii which are insensitive to cefazolin and cephaloridine. Pronounced activities were noted with 7-[2-(2-aminothiazol-4-yl)-acetamido]-3-[[[1-(2-dimethylaminoethyl)-1H-tetrazol-5-yl]thio]methyl]ceph-3-em-4-carboxylic acid (1y; SCE-963).


Membrane-bound penicillin-binding proteins of Escherichia coli. Comparison of a strain carrying an R-factor and the parent strain. Membrane-bound penicillin-binding proteins of an Escherichia coli carrying an R factor which mediated the resistance to penicillins were examined by slab gel electrophoresis and fluorography using beta-lactamase inhibitors such as methicillin, clavulanic acid and MC-696-SY2-A, and by affinity chomatography. By fluorography, it appeared that the penicillin-binding proteins of the strain carrying the R factor could not be distinguished from those of the parent strain. In both strains, methicillin had a preferential affinity for penicillin-binding proteins 2 and 3, clavulanic acid for 2 and 4, and MC-696-SY2-A for 1A at the concentration which was needed to inhibit about 75 approximately 80% of beta-lactamase activity of the membrane fraction from a strain carrying an R factor. This with other facts indicates that MC-696-SY2-A has a unique character in the binding to penicillin-binding proteins. By affinity chromatography using cephalexin-CH-Sepharose 4B column, two major cephalexin-binding proteins were detected. Their molecular weights were found to be 110,000 and 32,000, respectively. These two proteins correpsonded to penicillin-binding proteins 1 and 5/6. From these results it was suggested that the R factor had no influence on the penicillin-binding proteins in the E. coli strain examined.


Enzymatic studies on the mechanism of action of cefoxitin. Correlation between the affinities of cefoxitin to penicillin-binding proteins and its rates of inhibition of the respective penicillin-sensitive reactions in E. coli. The affinities of cefoxitin, a cephamycin antibiotic, to penicillin-binding proteins of Escherichia coli were reexamined using a recently developed method for separating penicillin-binding proteins. The inhibitions by this antibiotic of four measurable penicillin-sensitive enzymatic reactions, the reactions of D-alanine carboxypeptidases IA and IB, cross-bridge formation and concomitant release of D-alanine, were also measured. An approximate correlation was found between the affinities of cefoxitin to the penicillin-binding proteins responsible for these reactions and its rates of inhibition of the respective penicillin-sensitive reactions.


Determination of the ID50 values of antibacterial agents in agar. Microorganisms were plated on agar plates containing various concentrations of an antimicrobial drug and inhibition of growth was determined at each drug concentration. The ID50 value and the gradient of the line were calculated by the least square method. When 200 approximately 800 bacterial cells were inoculated on an agar plate, growth inhibition corresponded linearly with the log concentration of a drug within range of 5 approximately 95% inhibition. The ID50 value and the gradient obtained were reproducible and reliable using microorganisms at stationary phase of growth with all tested bacterial species and all tested antimicrobial agents. It was found that the ID50 values of drugs were more reproducible and may be more reliable than the MIC (minimum inhibitory concentration) values of the drugs.


The bacterial outer-membrane permeability of beta-lactam antibiotics. Two penicillins and 5 cephalosporins were evaluated for their ability to pass through the outer-membranes of Proteus morganii, Citrobacter freundii and Escherichia coli. Cefazolin, ceftezole and cephaloridine showed high permeability through the outer-membranes of these Gram-negative bacteria. Benzylpenicillin and cephalothin, on the contrary, showed low permeability. The outer-membrane permeability of ampicillin and cephalexin varied from species to species. C. freundii was found to have the highest barrier against both the penicillins and the cephalosporins, and E. coli appeared to have a low barrier against the cephalosporins. The hydrophobic character of the beta-lactam antibiotics, which was estimated by a reversed-phase thin-layer chromatography was closely related to the outer-membrane permeability. In general, the more hydrophilic antibiotic showed the higher outer-membrane permeability. However, cephaloridine, the most lipophilic compound among the antibiotics tested, showed good permeability.


The role of auditory stimuli in crying inhibition in the neonate. An experiment on the latency and duration of the cessation of spontaneous crying supported evidence that a low frequency intense auditory stimulus inhibits crying in neonates. There was a slight increase in the length of the first pause in crying during exposure to a 200-c/s tone, but no difference in the latency of this first pause. Since crying intensity reaches 80 db SPL at the infant ear, it is hypothesized that auditory stimuli juxtaposed during crying is in fact masked. Hence to be effective, an auditory stimulus must be presented at the beginning of a natural pause in crying. An experiment on rhythm in cry patterns did not support the hypothesis that crying may be a naturally rhythmic pattern of behavior analogous to sucking and susceptible to psychophysical investigation. There was a large variance within and between Ss for the mean latency and duration of cry bursts and pauses. Exper. III asked mothers with infants of 10 days, 7 weeks, and 13 weeks of age to keep a crying activity schedule in their own homes. This study revealed a discontinuity between neonates and older infants in the conditions and patterns of crying. Interview data did indicate that reliable positive orienting responses did occur to natural auditory stimuli in a naturalistic setting.


Proem to a quantum leap in audiometric data collection and management. A revolution began in 1961 when Weiss first mated a minicomputer to an audiometer. Today's technology offers a number of aids to the audiometrist in standardizing stimulus presentation patterns, objectifying and validating responses, computing HTL from a response pattern, computing significant threshold shifts from baseline audiograms, computing percentage binaural hearing impairment for a person, etc. Audiometers can be made which self-calibrate and are fail-safe. Furthermore, a revolution has occurred in replacing an audiogram written record, for example a card, with a system in which the audiometer transfers HTL data directly into computer storage without the possibility of transcription error. A group of turnkey and interlocking terminals allows the audiometric data of any individual serviced to be retrieved and/or updated from any terminal in the system. This paper lists commercial models simulating standard manual audiometry, models computerizing Bekesy audiometry, and some experimental models on yet other principles. Psychometric methods and methods for response validation suitable for audiometry are discussed, and out of many possible suggestions a few are offered for audiometric paradigms which may go beyond simulation of ASHA guidelines and ANSI specifications for manual threshold audiometry in order to take advantage of the power available from the computer as a tool in audiometry. For example, a computer can scan the responses of as many as 32 subjects simultaneously, offer each an adaptive program based on his response, and printout HTL's in conventional audiometric format.


Regulation of cell size in the yeast Saccharomyces cerevisiae. For cells of the yeast Saccharomyces cerevisiae, the size at initiation of budding is proportional to growth rate for rates from 0.33 to 0.23 h-1. At growth rates lower than 0.23 h-1, cells displayed a minimum cell size at bud initiation independent of growth rate. Regardless of growth rate, cells displayed an increase in volume each time budding was initiated. When abnormally small cells, produced by starvation for nitrogen, were placed in fresh medium containing nitrogen but with different carbon sources, they did not initiate budding until they had grown to the critical size characteristic of that medium. Moreover, when cells were shifted from a medium supporting a low growth rate and small size at bud initiation to a medium supporting a higher growth rate and larger size at bud initiation, there was a transient accumulation of cells within G1. These results suggest that yeast cells are able to initiate cell division at different cell sizes and that regulation of cell size occurs within G1.


Expression of RNA polymerase and ribosome component genes in Escherichia coli mutants having conditionally defective RNA polymerases. The expression of the genes coding for the beta and beta' subunits of RNA polymerase, ribosomal RNA, ribosomal proteins, and beta-galactosidase was investigated in strains carrying conditionally lethal mutations affecting either RNA polymerase core assembly or RNA polymerase enzyme activity. The mutant strain XH56 produces a temperature-sensitive beta' subunit and at 42 degrees C is defective in RNA chain initiation; consequently, little or no transcription occurs at the restrictive temperature. A partial restriction, produced by shifting the strain to 39 degrees C, resulted in a rapid fivefold increase in the transcription of the rpoB and C genes and in the synthesis of the beta- and beta'-subunit proteins for which they code. The RNA polymerase assembly-defective strains A2R7 and TS4 exhibited a 1.5- to 2-fold increase in the transcription of the rpoB and C genes and in the synthesis of beta- and beta-subunit proteins after prolonged restriction. These results demonstrate (i) that regulation of the synthesis of the beta- and beta-RNA polymerase subunits is under these conditions primarily transcriptional rather than translational, and (ii) that a stimulation of rpoB and C gene expression results from a restriction on RNA synthesis caused by either RNA polymerase inactivation or inhibition of its assembly. During restriction of the mutant strains, the transcription of the ribosome component genes exhibited patterns which were similar to transcription of the rpoB and C genes, supporting the evidence that genes coding for RNA polymerase are cotranscribed with ribosomal protein genes; transcription of the lacZ gene was observed to decrease concomitant with the stimulation of the rpoB and C genes.


Characterization of mutationally altered dihydropteroate synthase and its ability to form a sulfonamide-containing dihydrofolate analog. Among spontaneous mutants of Escherichia coli selected for resistance against sulfonamides, thermosensitive strains were found. These were shown to possess a changed dihydropteroate synthase (EC 2.5.1.15), which had a substantially higher Km value for its normal substrate, p-aminobenzoic acid, and an about 150-fold higher Km for sulfonamides. The mutationally changed dihydropteroate synthase was found to be thermosensitive by in vitro assays. The thermosensitivity was used as an enzyme marker to demonstrate the complex formation between 2-amino-4-hydroxy-6-pyrophosphorylmethyl pteridine and sulfonamides by partially purified dihydropteroate synthase. The formation of folate from 2-amino-4-hydroxy-6-pyrophosphorylmethyl pteridine and p-aminobenzoylglutamic acid by dihydropteroate synthase was found to be very sensitive to inhibition by sulfonamides and very inefficient with the mutationally changed enzyme.


Regulation of phenylalanine oxidase synthesis in Proteus mirabilis. Cells of Proteus mirabilis could oxidize L-phenylalanine to phenylpyruvate only when grown in the presence of a number of amino acids, particularly, L-alanine, L-asparagine, L-glutamate, and L-glutamine. Production of phenylalanine oxidase was slowly lost upon growth in a minimal medium containing ammonium ions as a nitrogen source but was reversed by the addition of casein hydrolysate. Oxidase activity as well as a phenylalanine-dichlorophenolindophenol (DCIP) reductase activity increased in P. mirabilis only during cell multiplication. Both rifampin and nalidixic acid caused inhibition of oxidase synthesis. A phenylalanine-active transport was found to be operative when bacteria were grown in the absence of added amino acids. After anaerobic growth, cells of P. mirabilis had lost their ability to carry the phenylalanine oxidase reaction when assayed in the presence of air, and nitrate could not be used as an electron acceptor for the oxidation of phenylalanine. However, some phenylalanine-dichlorophenolindophenol reductase activity was still present in anaerobic bacteria at the early stage of cell multiplication.


Inactivation of bacteriophages by protein E, a new major membrane protein isolated from an Escherichia coli mutant. Pure protein E, obtained after diethylaminoethyl-cellulose chromatography of ethylenediaminetetraacetic acid-Triton X-100-solubilized outer membrane proteins of Escherichia coli strain JF694, inactivated bacteriophage K3. Lipopolysaccharide enhanced bacteriophage inactivation. Antibody prepared against purified protein E protected bacteriophage K3 from inactivation by protein E. Bacteriophage K3 used a major outer membrane protein, protein II*, as part of its receptor. We conclude that proteins E and II* have a common region which interacts with bacteriophage K3. Protein E also inactivated two recently described bacteriophages, TC45 and TC23, that use protein E as at least part of their receptor.


Pyrimidine dimer excision in Escherichia coli strains deficient in exonucleases V and VII and in the 5' leads to 3' exonuclease of DNA polymerase I. An isogenic series of Escherichia coli strains deficient in various combinations of three 5' leads to 3' exonucleases (exonuclease V, exonuclease VII, and the 5' leads to 3' exonuclease of DNA polymerase I) was constructed and examined for the ability to excise pyrimidine dimers after UV irradiation. Although the recB and recC mutations (deficient in exonuclease V) proved to be incompatible with the polA(Ex) mutation (deficient in the 5' leads to 3' exonuclease of DNA polymerase I), it was possible to reduce the level of the recB,C exonuclease by the use of temperature-sensitive recB270 recC271 mutants. It was found that, by employing strains deficient in exonuclease V, postirradiation DNA degradation could be reduced and dimer excision measurements could be facilitated. Mutants deficient in exonuclease V were found to excise dimers at a rate comparable to that of the wild type. Mutants deficient in exonuclease V and the 5' leads to 3' exonuclease of DNA polymerase I are slightly slower than the wild type at removing dimers accumulated after doses in excess of 40 J/m2. However, although strains with reduced levels of exonuclease VII excised dimers at the same rate as the wild type, the addition of an exonuclease VII deficiency to a strain with reduced levels of exonuclease V and the 5' leads to 3' exonuclease of DNA polymerase I caused a marked decrease in the rate and extent of dimer excision. These observations support previous indications that the 5' leads to 3' exonuclease of DNA polymerase I is important in dimer removal and also suggest a role for exonuclease VII in the excision repair process.


Recipient competence in F'lac matings of Escherichia coli K-12. We studied recipient mating ability in the presence of excess F'lac donors. Ninety-five percent of recipients were able to receive F'lac in 30-min matings. Competition between an F'-lac donor and an F'lac traI donor, which mobilized a ColE1 derivative (pML2), showed that each recipient mated with an average of two to three donors in 30 min. Experiments in which the competing donor was added at different times showed that some competition occurred throughout the 30-min mating period, which suggested that aggregate formation was spread over this time.


Relationship of glycolytic intermediates, glycolytic enzymes, and ammonia to glycogen metabolism during sporulation in the yeast Saccharomyces cerevisiae. To identify the factors which control glycogen synthesis in Saccharomyces cerevisiae, we have studied the regulation of glycogen metabolism during sporulation, since in vivo glycogen has been reported to undergo significant changes in concentration during this process. We examined the concentration of a number of key glycolytic intermediates and enzymes in strains that sporulate at different rates and those that are deficient in sporulation. There were no significant changes found in the adenylate energy charge or cyclic AMP levels throughout sporulation. Although significant alterations occurred in the levels of glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-bisphosphate, phosphoenolpyruvate, and ATP during sporulation, only the fourfold increase in fructose-1,6-bisphosphate appeared to correlate with glycogen synthesis in all of the strains examined. Only limited changes occurred in the level of a number of glycolytic and gluconeogenic enzymes which were examined during this process. Intracellular glucose content underwent a dramatic 30- to 40-fold increase in sporulating cells. Comparison of strains with different rates of sporulation demonstrated that this increase in glucose content coincides with the time of glycogen degradation in each strain. Both the increase in glucose content and the degradation of accumulated glycogen were not observed in nonsporulating alpha/alpha strains, or in cells incubated in NH(4) (+) supplemented sporulation medium. Although glucose appears to be the direct product of glycogen degradation, a 10-fold increase in a nonspecific alkaline phosphatase occurs at this time, which may be degrading phosphorylated sugars to glucose. All of the strains examined released extracellular glucose while suspended in acetate sporulation medium. It is concluded that most of the changes in the glycolytic pathway that occur during sporulation, with the exception of glycogen degradation and the concomitant increase in intracellular glucose pools, are a response to the transfer to sporulation medium and are independent of sporulation-specific processes. Inhibition of sporulation with ammonium ions resulted in a different pattern of change in all of the glycolytic intermediates examined, including a twofold increase in cyclic AMP levels. Ammonia did not interfere with glycogen synthesis, but prevented sporulation-specific glycogen degradation. The levels of the glycolytic enzymes examined were not affected by ammonia.


Incorporation of phosphatidylglycerol into murein lipoprotein in intact cells of Salmonella typhimurium by phospholipid vesicle fusion. The biosynthesis of the diglyceride moiety of murein lipoprotein was studied by fusion of labeled phospholipid vesicles with intact cells of Salmonella typhimurium. Phosphatidylglycerol was found to be an excellent donor for the glyceryl moiety in lipoprotein, whereas phosphatidylethanolamine and cardiolipin were not. The incorporation of radioactivity from monoacyl-phosphatidylglycerol into lipoprotein can be attributed to its conversion to phosphatidylglycerol. The results strongly support our hypothesis that the glyceryl residue covalently linked to murein lipoprotein is derived from the nonacylated glycerol moiety of phosphatidylglycerol.


Escherichia coli K-12 mutants that allow transport of maltose via the beta-galactoside transport system. We have isolated mutants of Escherichia coli that have an altered beta-galactoside transport system. This altered transport system is able to transport a sugar, maltose, that the wild-type beta-galactoside transport system is unable to transport. The mutation that alters the specificity of the transport system is in the lacY gene, and we refer to the allele as lacYmal. The lacYmal allele was detected originally in strains in which the lac genes were fused to the malF gene. Thus, as a result of gene fusion and isolation of the lacYmal mutation, a new transport system was evolved with regulatory properties and specificity similar to those of the original maltose transport system. Maltose transport via the lacYmal gene product is independent of all of the normal maltose transport system components. The altered transport system shows a higher affinity than the wild-type transport system for two normal substrates of the beta-galactoside transport system, thiomethyl-beta-D-galactoside and o-nitrophenyl-beta-D-galactoside.


uvrC gene function in excision repair in toluene-treated Escherichia coli. We have examined the role of the uvrC gene in UV excision repair by studying incision, excision, repair synthesis, and DNA strand reformation in Escherichia coli mutants made permeable to nucleoside triphosphates by toluene treatment. After irradiation, incisions occur normally in uvrC cells in the presence of nicotinamide mononucleotide (NMN), a ligase-blocking agent, but cannot be detected otherwise. We conclude that repair incisions are followed by a ligation event in uvrC mutants, masking incision. However, a uvrC polA12 mutant accumulates incisions only slightly less efficiently than a polA12 strain without NMN. Excision of pyrimidine dimers is defective in uvrC mutants (polA(+) or polA12) irrespective of the presence or absence of NMN. DNA polymerase I-dependent, NMN-stimulated repair synthesis, which is demonstrable in wild-type cells, is absent in uvrC polA(+) cells, but the uvrC polA12 mutant exhibits a UV-specific, ATP-dependent repair synthesis like parental polA12 strains. A DNA polymerase I-mediated reformation of high-molecular-weight DNA takes place efficiently in uvrC polA(+) mutants after incision accumulation, and the uvrC polA12 mutant shows more reformation than the polA12 strain after incision. These results indicate that normal incision occurs in uvrC mutants, but there appears to be a defect in the excision of pyrimidine dimers, allowing resealing via ligation at the site of the incision. The lack of NMN-stimulated repair synthesis in uvrC polA(+) cells indicates that incision is not the only requirement for repair synthesis.


Characterization of amber and ochre suppressors in Salmonella typhimurium. Amber and ochre suppressor mutations in Salmonella typhimurium were selected. The amino acid insertions directed by the suppressors were inferred from suppression patterns of Escherichia coli lacI amber mutations. These amber mutations only respond to nonsense suppressors that direct the insertion of particular amino acids. Four Salmonella amber suppressors characterized insert serine, glutamine, tyrosine, and (probably) leucine. Of the three ochre suppressors characterized, two direct the insertion of tyrosine and one directs that of lysine. Of the three amber and two ochre suppressors which have been mapped by phage P22 cotransduction, all are located in the same relative position on the Salmonella map as the analogous E. coli suppressors are on the E. coli map.


Methylation-dependent DNA synthesis in Escherichia coli mediated by DNA polymerase I. An in vitro system was used to study DNA synthesis in lysates of Escherichia coli cells which had been grown in the presence of ethionine. Such lysates showed a reduced capacity to incorporate [3H]TTP into high-molecular-weight material. Activity could be restored by incubation with S-adenosyl methionine and ATP. S-adenosyl methionine-reactivated TTP incorporation required the presence of DNA polymerase I, ATP, and all four deoxyribonucleotide triphosphates. DNA polymerase III was not required.


Direct determination of the properties of peptide transport systems in Escherichia coli, using a fluorescent-labeling procedure. A direct study of peptide uptake by Escherichia coli was made using a fluorescent procedure. After incubation with the bacteria, peptides remaining in the medium were dansylated, separated chromatographically, and quantitated from their fluorescent intensities and/or from their incorporated radioactivity when tritiated dansyl derivatives were prepared. Peptide uptake was apparently not regulated and proceeded continuously until complete, with the absorbed peptides undergoing rapid intracellular hydrolysis and the excess amino acid residues leaving the cell. Thus, peptide uptake and amino acid exodus occur concurrently. However, peptidase-resistant substrates, e.g. triornithine and glycylsarcosine, which can be similarly estimated in cell extracts, were accumulated about 1,000-fold. The influence of amino acid composition and chain length on rates of transport was assessed. Different strains of E. coli showed variability in their rates of di- and oligopeptide transport. With respect to energy coupling, both the di- and oligopeptide permeases behaved like shock-sensitive transport systems.


Five different enzymatic activities are associated with the multienzyme complex of fatty acid oxidation from Escherichia coli. The purified multienzyme complex of fatty acid oxidation from Escherichia coli was found to possess 3-hydroxyacyl-coenzyme A (CoA) epimerase and cis-delta3-trans-delta2-enoyl-CoA isomerase activities in addition to the previously identified enoyl-CoA hydratase, L-3-hydroxyacyl-CoA dehydrogenase, and 3-ketoactyl-CoA thiolase activities. Evidence is presented in support of the proposed association of all five enzyme activities with one protein which apparently is composed of two types of subunits and which can exist in several aggregated forms. The five component enzymes of the complex were rapidly inactivated by tris(hydroxymethyl)aminomethane, whereas they remained active in the presence of potassium phosphate.


Penicillin-binding proteins in Proteus species. Penicillin-binding proteins in three species of Proteus, Proteus mirabilis, P. morganii, and P. rettgeri, were investigated by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. Penicillin-binding proteins in these Proteus species were compared with those in Escherichia coli K-12. An approximate correlation between penicillin-binding proteins in E. coli and those in Proteus species was shown by several criteria: electrophoretic mobilities; affinities of several beta-lactam antibiotics which show characteristic patterns of binding to penicillin-binding proteins in E. coli; relation between affinities of antibiotics to the proteins and effects on morphological changes in Proteus species; location of beta-lactamase activity among penicillin-binding proteins; and thermostability. The electrophoretic mobilities and several other characteristics of penicillin-binding proteins among the Proteus species examined were found to be similar from species to species and differed only slightly from those of E. coli.


Escherichia coli mutants overproducing phenylalanyl- and threonyl-tRNA synthetase. The structural genes for threonyl-tRNA synthetase (ThrRS) and phenylalanyl-tRNA synthetase (PheRS) are closely linked on the Escherichia coli chromosome. To study whether these enzymes share a common regulatory element, we have investigated their synthesis in mutants which were selected for overproduction of either ThrRS or PheRS. It was found that mutants isolated previously for overproduction of ThrRS as strains resistant to the antibiotic borrelidin (strains Bor Res 3 and Bor Res 15) did not show an elevated level of PheRS. PheRS-overproducing strains were then isolated as revertants of strains with structurally altered enzymes. Strain S1 is a temperature-resistant derivative of a temperature-sensitive PheRS mutant, and strain G118 is a prototrophic derivative of a PheRS mutant which shows phenylalanine auxotrophy as a consequence of an altered K(m) of this enzyme for the amino acid. In both kinds of revertants, S1 and G118, the concentration of PheRS and ThrRS was increased by factors of about 2.5 and 1.8, respectively, whereas the level of other aminoacyl-tRNA synthetases was not affected by the mutations. Genetic studies showed that the simultaneous overproduction of PheRS and ThrRS in revertants G118 and S1 is based upon gene amplification, since this property was easily lost after growing the cells in the absence of the selective stimulus, and since this loss could be prevented by the presence of the recA allele. By similar criteria, the four- and eightfold overproduction of ThrRS in strains Bor Res 3 and Bor Res 15, respectively, was very stable genetically, indicating that it is caused by a mutational event other than gene amplification. From these results, we conclude that the concomitant increase of PheRS and ThrRS in strains G118 and S1 is an expression of gene duplication and not of a joint regulation of these two aminoacyl-tRNA synthetases. This conclusion is further supported by the result that, in mutant G118 as well as in its parental strain G1, growth in minimal medium lacking phenylalanine led to an additional twofold increase of their PheRS concentration. This increase was restricted to the PheRS, since the level of other aminoacyl-tRNA synthetases, including the ThrRS, stayed unchanged.


ColE1 hybrid plasmids for Escherichia coli genes of glycolysis and the hexose monophosphate shunt. The Clarke-Carbon clone bank carrying ColE1-Escherichia coli DNA has been screened by conjugation for complementation of glycolysis and hexose monophosphate shunt mutations. Plasmids were identified for phosphofructokinase (pfkA), triose phosphate isomerase (tpi), phosphoglucose isomerase (pgi), glucose-6-phosphate dehydrogenase (zwf), gluconate-6-phosphate dehydrogenase (gnd), enolase (eno), phosphoglycerate kinase (pgk), and fructose-1,6-P2 aldolase (fda). Enzyme levels for the plasmid-carried gene ranged, for the various plasmids, from 4- to 25-fold the normal level.


Deletion analysis of the expression of rRNA genes and associated tRNA genes carried by a lambda transducing bacteriophage. Transducing phage lambdailv5 carries genes for rRNA's, spacer tRNA's (tRNA1 Ile and tRNA1B Ala), and two other tRNA's (TRNA1 Asp and tRNA Trp). We have isolated a mutant of lambdailv5, lambdailv5su7, which carries an amber suppressor mutation in the tRNA Trp gene. A series of deletion mutants were isolated from the lambdailv5su7 phage. Genetic and biochemical analyses of these deletion mutants have confirmed our previous conclusion (E. A. Morgan, T. Ikemura, L. Lindahl, A. M. Fallon, and M. Nomura, Cell 13:335--344, 1978) that the genes for tRNA1 Asp and tRNA Trp located at the distal end of the rRNA operon (rrnC) are cotranscribed with other rRNA genes in that operon. In addition, these deletions were used to define roughly the physical location of the promoter(s) of the rRNA operon carried by the lambdailv5su7 transducing phage.


Bacteriophage T4D receptors and the Escherichia coli cell wall structure: role of spherical particles and protein b of the cell wall in bacteriophage infection. The nature of the interaction of bacteriophage T4D and the outer cell wall of its host, Escherichia coli B, has been investigated. Bacteria with altered or modified cell walls have been obtained by two different growth procedures: (i) growth in high osmolarity medium or (ii) growth in broth in the presence of divalent heavy metal ions. When these altered host cells were washed and subsequently added to regular growth medium, they interacted with added phage particles, but successful infection did not occur. Most of the phage particles released from these treated cells were observed to have full heads and an altered tail structure. The altered phage tails had contracted sheaths and unusual pieces of the bacterial cell wall attached to the distal portion of the exposed phage tail tube. Phage released from bacteria grown in the high osmolarity medium had attached cell wall pieces of two major types, these pieces being either 40 or 21 nm in diameter. The smaller-type cell wall pieces (21 nm) were formed by three spheres each measuring 7 nm in diameter. Phage particles released from cells previously exposed to the divalent metal ions had only one 7-nm cell wall sphere attached to the distal end of the tail tube. It was found that these 7-nm spheres (i) are normal components of the cell wall and are morphologically similar to endotoxin, (ii) are held in place on the cell wall by a component of the cell wall called protein b, and (iii) are most likely the site of penetration of the phage tail tube through which the phage DNA enters the host cell.


Isolation and characterization of amber mutations in the lexA gene of Escherichia coli K-12. We describe the isolation and characterization of amber mutations in the lexA gene of Escherichia coli K-12. These mutations, designated spr(Am), were isolated and characterized in a lexA tif sfi genetic background. They abolished the sensitivity of the strain to UV light and resulted in high rates of synthesis of recA protein. Phage lambda+ failed to lysogenize the strains as observed with similar strains carrying non-amber spr mutations described previously, thereby indicating a constitutive expression of the phage induction pathway. Introduction of an amber suppressor mutation into a strain bearing the spr(Am) mutation restored expression of the LexA mutant phenotype. We conclude that spr mutations either inactivate or prevent synthesis of the lexA gene product and that loss of this product results in constitutive expression of the E. coli induction system in the tif sfi genetic background.


Localization of proteolytic activity in the outer membrane of Escherichia coli. An enzyme in the cytoplasmic membrane, nitrate reductase, can be solubilized by heating membranes to 60 degrees C for 10 min at alkaline pH. A protease in the cell envelope has been shown to be responsible for this solubilization. The localization of this protease in the outer membrane was demonstrated by separating the outer membrane from the cytoplasmic membrane, adding back various forms of outer membrane protein to the cytoplasmic membrane, and following the increase in nitrate reductase solubilization with increasing amounts of outer membrane proteins. This solubilization is accompanied by the cleavage of one of the subunits of nitrate reductase and is inhibited by the protease inhibitor p-aminobenzamidine. Analysis of membrane proteins synthesized by cells grown in the presence of various amounts of p-aminobenzamidine revealed that p-aminobenzamidine affects the synthesis of the major outer membrane proteins but has little effect on the synthesis of cytoplasmic membrane proteins. When outer membrane is reacted with the protease inhibitor [3H]diisopropylfluorophosphate, a single protein in the outer membrane is labeled. Since the interaction with diisopropylfluorophosphate is inhibited by p-aminobenzamidine, it is suggested that this single outer membrane protein is responsible for the in vitro solubilization of nitrate reductase and the in vivo processing of the major outer membrane proteins.


Organization of genes for transcription and translation in the rif region of the Escherichia coli chromosome. The lambdarifd18 transducing phage is known to carry several genes for components of transcriptional and translational machineries; these genes are clustered in the rif region at 88 min on the Escherichia coli genetic map. They include a set of genes for rRNA's (rrnB), a gene for spacer tRNA, tRNA2Glu (tgtB), one of the two genes for EF-Tu (tufB), genes for four ribosomal proteins (rplK, A, J, and L), genes for the beta and beta' subunits of RNA polymerase (rpoB and rpoC), and genes for three tRNA's (tyrU, gluT, and thrT). An additional tRNA gene (subsequently identified as thrU by Landy and his co-workers) and a gene for a protein (protein U) with unknown functions were found to be carried by lambdarif d18. We analyzed the organization of these genes by using various deletion and hybrid phages derived from lambdarif d18 and lambdarif d12, a phage related to lambdarif d18. The expression of various genes was examined in UV-irradiated cells infected with these transducing phages. Two main conclusions were obtained. First, the four tRNA genes are not cotranscribed with the genes in rrnB, even though these tRNA genes are located close to the distal end of rrnB. Second, the four ribosomal protein genes are organized into two separate transcriptional units; rplK and A are in one unit and rplJ and L are in the second unit. The first group of genes was shown to have a promoter separate from that for tufB or protein U. The second group of genes shares the promoter with rpoB and C, as described in a separate paper (M. Yamamoto and M. Nomura, Proc. Natl. Acad. Sci. U.S.A., 75:3891--3895). These and other results described in this paper show that the genes are organized in the following order: promoter, genes in rrnB; promoter, thrU, tyrU, (promoter?) glyT, thrT; (promoter?) tufB; promoter, a gene for protein U; promoter, rplK, rplA; promoter, rplJ, rplL, rpoB, rpoC.


Mutational evidence for identity of penicillin-binding protein 5 in Escherichia coli with the major D-alanine carboxypeptidase IA activity. The defect in D-alanine carboxypeptidase IA activity in the dacA11191 mutant of Escherichia coli was correlated with a defect in the release of penicillin G from penicillin-binding protein 5. The results suggest that penicillin-binding protein 5 catalyzes the major D-alanine carboxypeptidase IA activity of the wild type and that the mutation results in a defect in the deacylation step catalyzed by this enzyme.


rorA mutation of Escherichia coli K-12 affects the recB subunit of exonuclease V. The ATP-dependent nuclease, exonuclease V, of Escherichia coli plays an important role in repair and recombination. The enzyme is composed of two subunits, one of which is the product of the recB and recC genes. In this communication it is shown by mapping and complementation experiments that the rorA mutation, which results in radiation sensitivity but not the loss of recombination ability, is an allele of the recB gene.


Regulation of polyamine and streptomycin transport during stringent and relaxed control in Escherichia coli. Inhibition of polyamine uptake was observed during amino acid depletion in a stringent strain of Escherichia coli CP78 but not in a relaxed strain (CP79). Chloramphenicol was shown partially to relieve the inhibition of uptake. Stringent cells which were induced for a transport system common to both polyamines and streptomycin were found to restrict the uptake of spermidine as well as streptomycin.


Two-component nature of bacteriophage T4 receptor activity in Escherichia coli K-12. Escherichia coli bacteriophage T4 uses the lipopolysaccharide of the outer cell envelope membrane as a receptor. Lipopolysaccharide from E. coli K-12 required a major outer membrane protein, polypeptide Ib, for phage inactivation.


Outer membrane of Serratia marcescens: apparent molecular weights of heat-modifiable proteins in gels with different acrylamide concentrations. The major proteins from the outer membrane of Serratia marcescens SM-6 are heat modifiable. The analysis of their apparent molecular weights in gels with different concentrations of acrylamide and the results obtained by radioactive labeling indicate that the major proteins are covalently linked to carbohydrate moieties.


Method for isolating restriction- and modificationless mutants of Escherichia coli K-12. A simple method is described for the selection and isolation of restriction- and modificationless mutants in Escherichia coli K-12 by using the following properties: (i) the temperature-sensitive repressor activity of phage lambdacI857; (ii) a mutant of lambda phage defective in integration and the establishment of repression (lambdab2cI); (iii) a virulent lambda phage insensitive to the repressor activity. The final yield of spontaneously arising rk-mk+ and rk-mk- mutants from stationary-phase cultures was about 5% of the surviving cells.


Relationship between phosphate content and serological activities of the mannans of Candida albicans strains NIH A-207, NIH B-792, and J-1012. The mannans from Candida albicans strains NIH A-207 (serotype A), NIH B-792 (serotype B), and J-1012 (serotype C) were fractionated on a column of diethylaminoethyl-Sephadex into five subfractions containing different amounts of phosphate. Antibody-precipitating activities of the mannan subfractions of strains NIH A-207 and NIH B-792 were proportional to their phosphate content, while those of strain J-1012 did not show regularly proportional precipitin activity. A similar tendency was also observed in the cross-reaction between the mannan su,fractions of strains NIH A-207 and J-1012 and their heterologous antisera. The mannans of strain NIH B-792 showed lower cross-reactivities against antisera of strains NIH A-207 and NIH B-792, i.e., only two subfractions containing larger amounts of phosphate were able to react with these antisera.


Simple method for identification of plasmid-coded proteins. Proteins encoded by plasmid DNA are specifically labeled in UV-irradiated cells of Escherichia coli carrying recA and uvrA mutations because extensive degradation of the chromosome DNA occurs concurrently with amplification of plasmid DNA.


Accumulation of peptidyl tRNA is lethal to Escherichia coli. A mutant strain of Escherichia coli with temperature-sensitive peptidyl-tRNA hydrolase grows at 30 degrees C but, when shifted to 40 degrees C, dies at rates affected by physiological, pharmacological, and genetical perturbations. The rate of killing correlates with the relative accumulation of peptidyl-tRNA, suggesting that it is responsible for the death of the cells.


Conservation of primary structure of the pyridoxyl peptide of Escherichia coli and Serratia marcescens tryptophan synthase beta2 protein. Two labeled peptides were recovered from tryptic digests of the NaB3H4-reduced, performic acid-oxidized beta2 protein of Serratia marcescens tryptophan synthase. These two pyridoxyl peptides were identical except for the presence or absence of an NH2-terminal arginyl residue. Tryptic digestion of nonreduced, performic acid-oxidized protein allowed isolation of the peptides that comprise the two halves of the pyridoxyl peptide. The partial primary structure for this region of the protein was shown to be Arg-Glx-Asx-Ler-Leu-His(Gly,Gly,Ala,His)Lys(Pxy)-Thr-Asx-Glx-Val(Leu,Gly,Glx,Ala,Leu,Leu,Ala)Lys. All the data available indicate that the sequence is identical with the homologous region from the Escherichia coli enzyme.


Structural and functional analysis of cloned DNA segments containing the replication and incompatibility regions of a miniplasmid derived from a copy number mutant of NR1. A 1.45-megadalton segment of DNA cloned from a miniplasmid derived in vivo from a copy number mutant of the R plasmid NR1 has been shown to contain all functions essential for incompatibility and autonomous plasmid replication in Escherichia coli. Specific endonuclease cleavage sites within this DNA segment that localize functions required for replication have been mapped. A 0.45-megadalton fragment that specifies the FII incompatibility of NR1 has been identified within the replication region, and DNA fragments containing this incompatibility region, but lacking other functions required for replication, have been cloned.


Controlled comparison of trebenzomine with doxepin as an antidepressant-anxiolytic. Trebenzomine, a new psychotropic drug, was compared with doxepin in the treatment of anxiety and depression. The study was a three-week, double-blind trial involving 26 psychoneurotic inpatients. Multiple ratings by physician, nurse, and patient all concurred that the 2 drugs produced significant improvement in both depressive and anxious symptomatology. No significant differences were found between the 2 drugs either in terms of rate or degree of improvement or incidence of side effects. Further investigation of trebenzomine for anxious depression is warranted.