The mating behaviour of the tsessebe (Damaliscus lunatus lunatus) in the Kruger National Park. The mating behaviour of tsessebe was observed in the Kruger National Park. A description of the typical sequences in the mating behaviour is given which includes the "nose and neck stretch" posture of the courtship display of the bull when approaching a cow, the role of olfactory stimuli which are derived from either smelling at the base of a cow's tail or from the ground where a cow had been laying down or urinated, the spontaneous ejaculation of the bull in response to such stimuli and mounting. Distinction is drawn between the dominance display ("nose forward-upward posture") and the courtship display of the tsessebe bull. Comparative notes on the mating behaviour of congeneric species, i.e. the blesbok (D. dorcas phillipsi), bontebok (D. d. dorcas) and topi (D. lunatus korrigum) are given. Many similarities are evident despite a number of dissimilarities. Attention is also paid to confliction interpretations of the courtship displays in the different species.


Uganda kob (Adenota kob thomasi): territoriality and the spatial distributions of sexual and agonistic behaviors at a territorial ground). Possible functions of territorial behavior were evaluated on the basis of observations of agonistic and reproductive activities of Uganda kob at a "territorial ground" (TG) in southwestern Uganda. We have found that male residents of a TG tend to occupy exclusive areas, and that agonistic interactions are concentrated along the boundaries separating adjacent occupied areas. Thus, we conclude that male kob do engage in the active defense of spatially exclusive territories at a TG.


Scent marking by male Golden Hamsters (Mesocricetus auratus) III. Behavior in a seminatural environment. 8 malexj golden hamsters were observed two at a time in a plexiglas cage (area roughly 11 sp.ft) subdivided into 47 connected compartments. Each pair spent 2--3 weeks in this cage, observations lasting 1--2 weeks. Dominance relationships, food hoarding and marking behaviour in this situation are described.


First isolations of Levinea-Citrobacter cultures in Czechoslovakia. Twenty-two strains corresponding by their biochemical properties to the genus Levinea - Citrobacter were isolated. Six of the strains were referred to the species Citrobacter diversus and 12 to C. freundii, whose properties are identical with those of L. malonatica and L. amalonatica, respectively. Four strains differed from Levinea organisms by some reactions, but were fully compatible with C. freundii (in the scheme of EWING and DAVIS); two of them utilized malonate. The taxonomic position of strains displaying the following biochemical properties: dextrose positive, indole positive, H2S negative, urease on Christensen's citrate medium positive, lysine-decarboxylase negative-is discussed. In routine practice, these strains may be more accurately identified by adding of four tests: adonitol with gas production, KCN, raffinose and malonate.


Bone canaliculus endings in the area of the osteocyte lacuna. Electron-microscopic studies. Light- and electron-microscopic studies were carried out on the bone canaliculi and their endings in the wall fo the osteocyte lacuna. Two types of canalicular endings were distinguished in the lacuna wall. Those of the first type, fairly numerous, are the endings of small diameter, which do not branch off in the immediate proximity of the lacuna. The other type, occurring in twos or threes, are large in diameter and branch off in the close vicinity of the lacuna, invariably possessing two or three processes which run up to the point of branching of the canaliculus. The images obtained made it possible to demonstrate that the collagen fibrils in both types of endings in the lacuna wall are, in principle, arranged in a similar manner. Nonethless, the type of end formation found where the fibrils run along the axis of the lacuna differs from that for the fibrils in a plexus.


Study of alternative substrates in the catalytic reaction of N-acetyl-transferase. In the enzymatic acetylation of para-nitroaniline the esters of para-amino-benzoic acid act as inhibitors since they serve as alternative acceptor substrates. The extent of inhibition is increased by increasing the number of carbon atoms of the alcohol moiety of the esters. The apparent Km value of esters as acceptor substrates decreases by almost one order of magnitude from 1.1 times 10- minus 4 to 2.5 times 10- minus 5 M within the methylester-butylester series. It has been shown by kinetic analyses that the esters are non-competitive inhibitors in respect of the acetyl donor, whereas they are competitive inhibitors in respect of the acetyl acceptor, in the acetylation reaction of paranitroaniline.


Alkylmercurial encephalopathy in the monkey (Saimiri sciureus and Macaca arctoides): a histopathologic and autoradiographic study. Histopathologic and autoradiographic studies were performed on monkeys of the genera Saimiri and Macaca after acute and chronic oral exposure to several dosage regimens of methylmercuric chloride (MeHg). Neuropathologic changes were primarily cortical, although subcortical lesions also were observed. Autoradiographic localization of 203-Hg was greatest within glial cells (particularly Nissl-pump astrocytes, subependymal glia and Bergmann's glia) and mast cells. High levels of label within normal appearing large neurons (particularly those within Gasserian and dorsal root ganglia) indicate a lower susceptibility of these neurons to the toxic effects of MeHg. Blood and brain levels of mercury correlated well with the degree of neuropathologic change, but individual variations in susceptibility to intoxication also existed.


Incidence of congenital heart disease in Hungary: Pécs and County Baranya. A study has been made of the incidence of congenital heart disease in Pécs and County Baranya. A total of 744 affected individuals were found among 97 482 live-born infants examined during the 15-year period 1955 to 1969; this amounts to an incidence of 7.6 per thousand.


Spread of tuberculosis from obstructed and non-obstructed upper urinary tract. As part of an experimental study of the spread of urogenital tuberculosis, male guinea pigs were inoculated with tubercle bacilli (H37Rv) in the upper part of the ureter. Half of the animals were prepared by ligature of the ureter below the injection site prior to inoculation. Spread of the infection to the lower urinary tract and the genitalia occurred whether the ureter was intact or ligated. It is concluded that lymphatic spread is possible and seems to be more common than urinary (canalicular) spread.


Photically induced epilepsy in Papio papio as a model for drug studies. Neurophysiological studies employing drugs have been undertaken in the natural syndrome of photically induced epilepsy in the Senegalese baboon Papio papio. GABA-mediated inhibition, both pre- and posysynaptic, plays an important role in the epileptic manifestations seen in this syndrome synapses can significantly modify photically induced epileptic responses, partly as a result of changes in afferent activity. The level of activity in dopaminergic systems can also modify the epileptic signs. Among anticonvulsant drugs, barbiturates and benzodiazepines are very effective against this type of epilepsy, whereas many other drugs are weakly active or toxic. A modification of the natural model (using allylglycine as a priming agent) is convenient for correlating acute anticonvulsant activity and neurological toxicity with plasma concentrations of anticonvulsant agents.


Effect of chronic amphetamine exposure on stereotyped behavior: implications for pathogenesis of l-dopa-induced dyskinesias. The effect of chronic high doses of d-amphetamine on amphetamine- and apomorphine-induced stereotyped behavior in guinea pigs was studied in an attempt to determine the mechanism involved in the development of L-DOPA-induced dyskinesias in parkinsonism. Prolonged pretreatment with suprathreshold doses of amphetamine decreased the threshold dosage of both amphetamine and apomorphine necessary to elicit stereotyped behavior. This suggests that chronic striatal dopamine stimulation may induce striatal dopamine receptor site hypersensitivity  and that this type of induced hypersensitivity may play a role in the development of L-DOPA-induced dyskinesia. This mechanism may also play a role in the development of amphetamine-induced chorea and psychosis.


Apomorphine and piribedil in rats: biochemical and pharmacologic studies. We studied the biochemical and pharmacologic modes of action of piribedil and apomorphine in the rat. Although both drugs have many points in common, they are also different in many of their manifestations. Apomorphine causes high-intensity, short-duration stereotyped behavior; it is distributed within the brain in uneven fashion, the striatum being the area of lowest concentration as measured by fluorometry. Direct stereotactic injection within the dopaminergic mesolimbic system, and particularly the tuberculum olfactorium, produced constant intense responses. All effects of apomorphine can be blocked by pimozide, but propanolol, a beta blocker, only reduces aggression and ferocity, leaving stereotyped behaviors intact. Finally, L-5-HTP tends to reduce aggression, ferocity, and to a lesser extent stereotypy; MIF or piribedil, as well as reserpine, potentiates the stereotyped behaviors induced by apomorphine, whereas L-DOPA usually decreases them. Piribedil, on the other hand, causes low-intensity, long-duration stereotyped behavior. It is distributed within the brain almost uniformly. Most effects of piribedil can be blocked by pimozide, but propanolol blocks only aggression and ferocity, leaving stereotyped behaviors intact. On the other hand, clonidine, an alpha-receptor agonist, blocks stereotyped behaviors induced by piribedil but markedly increases aggression, ferocity, and motor activity. L-5-HTP and L-DOPA have little effect on piribedil-induced manifestations. Reserpine decreases piribedil stereotypy. The main metabolite of piribedil, S 584, had no clear-cut pharmacologic action in our hands at the dosage used. It is concluded that both apomorphine and piribedil produce stereotyped behavior by modifying the physiologic balance between mesolimbic and nigrostriatal dopaminergic systems. The other actions of apomorphine and piribedil upon aggression, ferocity, and motor activity are not always in parallel and depend probably on the fact that piribedil is less specific, affecting also noradrenergic, serotonergic, histaminergic, and/or cholinergic circuits. The usefulness of piribedil against some forms of human tremor and its low-intensity antiakinetic action probably result from this pattern of pharmacologic activity.


Retrograde catheterization of the left atrium in idiopathic hypertrophic subaortic stenosis. Retrograde catheterization of the left atrium utilizing the Shirley technique was successfully performed in four patients with idiopathic hypertrophic subaortic stenosis. By traversing the inflow tract of the left ventricle on pullback from the left atrium to the aorta, this method served to separate definitively true hypertrophic subaortic stenosis from cavity obliteration. It was accomplished with relative ease and no complications. In patients with suspected idiopathic hypertrophic subaortic stenosis who are to have retrograde left heart catheterization, this approach to the left atrium is recommended as an alternative to the transseptal technique.


A scanning electron microscopic study of the luteo-follicular complex. II. Events leading to ovulation. Morphological changes on the ovarian surface of different mammals both before and during ovulation have been examined by scanning electron microscopy. Preovulatory follicles were blisterlike structures that protruded markedly from the ovarian surface. Basal areas of preovulatory follicles were covered with polyhedral cells containing numerous microvilli, whereas on the lateral surfaces, superficial cells were elongated and possessed few microvilli. At the apex of the follicle, cells were very flattened and possessed few microvilli, which were present only in regions of intercellular contact. In some apical areas, cells appeared to be degenerating, whereas in other regions, groups of cells had "sloughed off." In addition, a fluidlike material was observed to exude from intercellular spaces of the superficial epithelium and to cover some apical cells. By transmission electron microscopy, the same fluidlike material was observed to (1) infiltrate the connective tissue of the tunica albuginea, (2) accumulate under the basal lamina, and (3) distend intercellular spaces of the superficial epithelium. Just prior to ovulation, large, irregular areas of the apex were ruptured and the oocyte, covered with a large amount of fluid, appeared to emerge from the follicle. At ovulation, the oocyte was not completely covered with follicle cells and the zona pellucida was clearly evident. The surface of the zona was quite irregular and contained numerous infoldings, channels and crypts. Follicle cells had polyhedral or star shapes and possessed large cytoplasmic evaginations that obliquely penetrated the zona. Both the zona pellucida and corona cells were covered with a fine layer of granular material. The SEM results and parallel TEM observations suggest that a local increase of fluids (edema) may be an important factor in the final decomposition of the distended and weakened apex of the preovulatory follicle. In addition, the participation of follicle cells, smooth muscle cells and the oviduct in the escape of the oocyte from the ruptured follicle is discussed.


Repeat ectopic pregnancy: a study of 123 consecutive cases. In a study of 1,330 ectopic pregnancies, 123 (9 per cent) patients experienced repeat tubal pregnancies. Of these 123 patients with repeat ectopics, 96 (78 per cent) were surgically sterilized with their second operative procedure. The remaining 27 potentially fertile patients have had nine pregnancies, but only three living children. Cornual wedge resection of the uterine tube predisposed in interstitial pregnancy and to rupture of the uterus in four cases. Four patients had three ectopic pregnancies each. Repair of the involved tube is usually technically possible following linear salpingotomy and should be done if future fertility is desired. If the patient does not desire future pregnancy, she should be sterilized to avoid the risk of repeat ectopic pregnancy.


Transfer of lidocaine across the sheep placenta to the fetus. Hemodynamic and acid-base responses of the fetal lamb. Placental transfer of lidocaine administered to pregnant sheep and removal of the drug from the fetal circulation were determined. Responses of the hemodynamic and acid-base state in the mother and fetus were also studied. Lidocaine readily crossed the placenta and appeared in the fetal blood as early as one minute following injection. Disappearance of lidocaine in the asphyxiated fetus tended to be slower than in the normal one. Lidocaine produced a transient fall in the fetal heart rate which was accompanied by a decrease in blood pH and oxygenation. Lidocaine also reduced umbilical blood flow and, in some instances, uterine blood flow. Decrease in umbilical blood flow was more pronounced in the initially asphyxiated fetus.


Maturational changes in the fetal glomerular filtration rate. Few data are available documenting the evolution of glomerular filtration rate (GFR) during gestation. In order to investigate this aspect, 33 fetal sheep have been studied, with the use of an intrauterine fetal preparation. It appears that the actual GFR (AGFR) in milliliters per minute increased during the last third of gestation without a significant increase in GFR expressed as milliliters per minute per kilogram of fetal weight (GFR-FW) and as milliliters per minute per gram of kidney weight (GFR-KW). It is suggested that a parallel and constant increase in AGFR and in total fetal weight or kidney weight might explain the absence of variation if GFR-FW or GFR-KW during the last third of gestation.


Use of radioactive microspheres for studies of tubal ovum transport. Experiments were designed to ascertain the fidelity with which radioactive plastic ovum models imitated the time-course of transport of natural ova through the rabbit oviduct. After location of the radioactive microspheres within the intact oviducts, the genital tracts were removed and cleared in benzyl benzoate. Natural ova and ovum surrogates were then visualized in situ. The radioactive microspheres were reasonably good models of transport of natural ova and their sojourn in the oviduct was shortened by progesterone and prolonged by estrogen. Thus they should be useful tools for future studies of own transport under various conditions.


Plasma estrogens in euthyroid and thyrotoxic women. Plasma levels of estrogen were measured daily by radioimmunoassay for 28 consecutive days in 12 healthy euthyroid women and 15 thyrotoxic women (10 with hypomenorrhea and 5 with amenorrhea) before commencement of therapy (except in one case) in an effort to increase understanding of the relationship between the thyroid gland and ovarian function. The results show that the patterns of plasma estrogen in thyrotoxic women with hypomenorrhea were similar to those in euthyroid women, although the levels in the thyrotoxic women were significantly elevated (P less than 0.001). Thyrotoxic women with amenorrhea also had a markedly elevated plasma estrogen level with a persistence of a sharp peak of estrogen but without the secondary estrogen rise associated with corpus luteum function. This suggests a failure of the positive feedback effect of estrogen on the hypothalamopituitary axis with resultant failure of ovulation.


Keratomycosis due to an Alternaria species. A patient with a contributory history of stromal herpes presented with a severe hypopyon and multiple corneal infiltrates. Culture of corneal scrapings grew out a species of Alternaria, a generally nonpathogenic fungus. The presence of hyphae and budding yeast forms in the keratoplasty specimen gave further confirmation to the diagnosis.


Psychiatric problems of children seen in an urban center of western India. This paper outlines the types of problems seen in a child guidance clinic in Western India. An attempt is made to understand the emotional disorders in the context of major socio-psychological changes taking place in a social order passing through a transitional stage from a traditional agrarian structure to an industrial state.


Prolyl hydroxylase and collagen metabolism after experimental mycardial infarction. Mature male Sprague-Dawley rats were subjected to an isoproterenol-induced myocardial infarction. Animals were sacrificed on a daily basis in order to assess the temporal changes in prolyl hydroxylase activity and collagen metabolism during the acute stages of myocardial necrosis and repair. Total myocardial hydroxyproline, as an indexof collagen content, increased promptly and markedly, beginning on day 4, and remained elevated thereafter. The incorporation of (14Cl)-proline into definitive hydroxyproline of mature collagen was also increased. The activity of the enzyme prolyl hydroxylase, which regulates the rate of conversion of proline to hydroxyproline in collagen, was elevated by day 2, remained high through day4, and then declined to a relatively constant but still slightly elevated level  throughout the period of repair. It is believed that changes in these parameters of collagen metabolism reflect changes in myocardial fibroblastic cell and ground substance pertinent to fundamental aspects of repair of the injuried myocardium.


Neutral amino acid transport pathways in uptake of L-thyroxine by Ehrlich ascites cells. Neutral amino acid pathways were investigated as possible mediators of L-thyroxine (T4) entry into ascites cells. 14C-Labeled T4, alpha-aminoisobutyric acid (AIB), L-phenylalanine (Phe), and cycloleucine (cLeu) uptakes were measured at initial extracellular amino acid concentrations of 1 muM in a modified Krebs-Ringer phosphate buffer, pH 7.4. T4 uptake was markedly slower than that of AIB, Phe, or cLeu over a 25-min period. T4 uptake was neither competitively stimulated nor inhibited by either AIB or Phe when these latter amino acids were varied over the concentration range 1.0 x 10(-6) to 1.0 x 10(-2) M. Preloading cells with 30 mM L-methionine did not enhance the uptake of T4. TJ efflux from preloaded cells was not affected by either AIB or Phe in the extracellular fluid (ECF). The uptakes of AIB, Phe, and cLeu were markedly altered by replacement of ECF Na+ with choline+, whereas T4 uptake was unchanged. It was concluded that neural amino acid transport pathways (specifically A, L, ASC, and beta) do not participate in the transmembrane transfer of L-thyroxine into Ehrlich ascites cells.


PSRO: advantages, risks, and potential pitfalls. The authors propose that active physician participation in current review methodologies, including PSRO, can lessen the potential for such problems as the development of unrealistic standards, emphasis on review and record keeping as ends rather than means, and ineffectuality of the systems themselves. Potential advantages of PSRO are seen in areas of quality assessment and improvement, economy, and enhancement of the continuing learning processess of physicians. The authors suggest that physicians should resist pressures toward separation of reviewing authority from clinical competence.


A process of establishing norms for inpatients length of stay in a community mental health center. A pervasive sense of urgency, stimulated by the Social Security Amendments of 1972 (Public Law 92-603), attends the need to establish norms for length of stay for impatient services in mental health facilities and to provide review mechanisms for extended care cases. The authors describe one model, adaptable to changing needs and federal regulations, that can offer a beginning experience in this endeavor.


A methodologic approach to crisis therapy. Resistance to using limited goal, brief psychotherapy often rests upon the mythology that a patient's problems must require a great deal of time for resolution. This paper offers a suitable rationale and methodology for psychotherapists to exercise their enormous capacity to assist patients effecting meaningful change in a brief time period.


Adverse reactions to intravascularly administered contrast media. A comprehensive study based on a prospective survey. Case reports on 112,003 patients obtained through a prospective survey have been studied and submitted to computer processing. The significant findings have been reviewed and their importance discussedmthe following points are noteworthy: the value of pretesting is doubtful; and routine pretesting is not a recommended procedure. The constant readiness of a fully equipped emergency tray offers far greater security and assurance of patient safety than the unpredictable and unreliable results of pretesting. The over-all incidence of nonfatal reactions is lower than has been prviously indicated: 5.65 per cent for intravenous urography; and 2.33 per cent for intravascular studies. The incidence of fatal reaction (1 in 10,000) is higher than that reported on the basis of retrospective studies. This figure may or may not be duplicated in the next 100,000 examinations. The cause of death in some patients may be related to a combination of factors-primary disease, diagnostic procedure, and contrast medium. The over-all incidence of adverse reactions in patients with allergy is about twice that in the general population. The incidence of adverse reactions is highest in the third and fourth decades, and lowest at either end of the age spectrum. Incidence of reactions is equal in both sexes. History of reaction to previous examinations is not a contraindication to re-examination. The incidence is approximately 3 times that of the general population. Rapid injection rate, in intravenous urography, is accompanied by fewer reactions than a slow injection rate. A slow injection rate in intravenous cholangiography is accompanied by fewer ractions than a rapid injection rate. It is anticipated that this program will continue, so that additional meaningful data and significant information will be accumulated. The members of our Committee are of the firm belief that this program will become the foundation of a permanent national and international adverse reaction reporting system, thus meeting a great need. We invite the collaboration and support of all those interested and involved in the many phases of contrast medium work.


Development of the tibial tuberosity.

Development of the tibial tuberosity. The tibial tuberosity develops by a concomitant, and juxtaposed, outgrowth of hyaline cartilage and ingrowth of fibrovascular tissue. During the fetal period this hyaline cartilage outgrowth is progressively distally displaced to become situated adjacent to the anterior portion of the tibial metaphysis. About four to six months post-natally a growth plate develops under the tibial tuberosity. This growth plate shows structural adaptations that permit accommodation of large tensile stresses. The growth plate of the tibial tuberosity has three distinct regions--(a) a zone of endochondral bone formation, (b) a zone of intramembranous bone formation through fibrocartilage, and (c) a zone of intramembranous bone formation through fibrous tissue.


Isoflurane anesthesia for pheochromocytoma: A case report. During an exploratory laparotomy for a pheochromocytoma under isoflurane anesthesia, the blood pressure of the patient rose to 250/115 torr, without ventricular arrhythmias. The authors consider isoflurane worthy of further study for pheochromocytoma-surgery anesthesia, in comparison with other presently used agents.


Effect of cocaine administration on halothane MAC in dogs. Intravenous cocaine (2 mg/kg or 4 mg/kg) was associated with a dose-dependent increase of halothane MAC in the dog. The largest increase in halothane MAC was from 1.06 plus or minus 0.03 to 1.35 plus or minus 0.04 vol/100 ml 3 hours after 4 mg/kg of cocaine. Halothane MAC 24 hours following a single administration of either 2 mg/kg or 4 mg/kg of cocaine or after 10 daily doses of 4 mg/kg of cocaine was near control MAC. These data support the speculation that increased central nervous system catecholamine concentrations, as may occur with cocaine, increase halothane anesthetic requirements.


Plasma concentration of lidocaine after endotracheal spray. The rate of absorption and plasma concentrations of lidocaine were determined in 15 supine patients under general anesthesia following either endotracheal spray of 200 mg. (average 3.3 mg./kg.) of 4 percent lidocaine hydrochloride solution with a special cannula (LTA Kit, Abbott) (N = 6) or 1 mg./kg. of 2 percent lidocaine solution as an intravenous bolus injection (N = 9). In the 6 patients after endotracheal spray, peak plasma lidocaine levels were reached at 15 to 20 minutes (mean 3.54 plus or minus 0.76 S.E. mug./ml. at 20 minutes). In the 9 patients given 1 mg./kg. intravenously, dose peak concentrations were reached within 2 minutes (mean 5.63 plus or minus 0.18 S.E. mug./ml. at 2 minutes). Plasma concentrations of lidocaine in both groups were within nontoxic ranges.


Humidification: Indications and hazards in respiratory therapy. Scientific data fail to support the widespread use of humidity and mist in respiratory therapy. A clear indication exists to provide humidification of inspired air in patients in whom the proximal airway is bypassed by endotracheal cannulation. Indications for the use of concentrated mist atmospheres in therapy of lung disease is less well documented, and too often the importance of adequate systemic hydration is overlooked. Theory supports the use of a highly humid or mist atmosphere in therapy of upper airway disease, but little benefit is derived from such therapy in diseases of the smaller airways and lung parenchyma. Danger of bacterial contamination and infection and an increase in airway resistance are significant hazards to the patient exposed to highly humid atmospheres.


Residual halothane: Patient and personnel exposure. In addition to offering further data on the exposure of surgical and recovery-room personnel to low levels of halothane in room air, the authors demonstrate the presence of halothane levels in the blood of some surgical patients managed by other inhalation agents from machines previously used for halothane. The authors concluded that these levels, while low, may be of clinical significance in selected patients.


Corneal injuries during general anesthesia. A corneal abrasion is the most common eye complication during general anesthesia and recovery. It is painful, may progress to inflammation of the uveal tract, and in the presence of contamination may lead to a serious infection. Both eyes may be involved, as the two reported cases show. Possible causes include mask friction on the open eye or other careless technics. Treatment includes local application of an antibiotic ointment and an eye pressure patch. In addition, a cycloplegic and mydriatic solution is instilled to prevent synechiae (in the presence of a secondary iridocyclitis) and to relieve the pain associated with spasm of the iris and ciliary muscle. Topical application of local anesthetics should be avoided, as they delay regeneration of corneal epithelium and may promote keratitis.


Thiopental and succinylcholine: Action on intraocular pressure. Intraocular pressure (IOP) measurements were made in a series of 92 male surgical patients, to assess the effects of timing and dosage of succinylcholine given after a standardized sleep dose of thiopental (3 mg./kg.). The major findings of this study were as follows: (1) thiopental alone lowered IOP; (2) a small (0.5 mg./kg.) dose of succinylcholine, given immediately after thiopental, returned IOP to normal; (3) a large (1 mg./kg.) dose of succinylcholine immediately after thiopental maintained the IOP at a low value; (4) if 2 minutes elapsed between thiopental and 1 mg./kg. of succinylcholine, the relaxant raised the IOP to slightly above preanesthetic control values; (5) tracheal intubation caused a significant rise in IOP, more than any effect from succinylcholine itself; (6) succinylcholine drip (0.1 percent), begun after establishment of satisfactory endotracheal halothane-nitrous oxide anesthesia, caused significant IOP elevation in 4 of 11 patients.


Ventricular arrhythmias after epinephrine injection in enflurane and in halothane anesthesia. The use of subcutaneous epinephrine during anesthesia is a common clinical practice for providing surgical hemostasis. In studies with 100 patients given either enflurane or halothane, with or without subcutaneous epinephrine, the incidence of ventricular ectopy in patients receiving halothane without epinephrine was 3 percent, while in those given epinephrine with halothane, the incidence was 7 percent. Those who received enflurane alone had no ectopic beats, while ventricular ectopy with enflurane and epinephrine resulted in an incidence of 1 percent. The authors conclude that enflurane anesthesia with concomitant administration of subcutaneous epinephrine is safe, provided the safeguards previously established for use of epinephrine with halothane are observed.


Intraocular pressure changes during anesthesia for electroshock therapy. Intraovular pressure was measured in 20 patients undergoing anesthesia for electroshock therapy. Compared to control values, determined on the day before electroshock therapy, statistically significant elevations of intraocular pressure followed the post-electroshock therapy convulsions and were still significant 5 minutes later, but not of sufficient magnitude or duration to cause concern.


Effects of hemodilution on hypotention during cardiopulmonary bypass. Arterial pressure, systemic vascular resistance, and viscosity measurements were made on 20 patients, during cardiopulmonary bypass with hemodilution to an average hematocrit reduction of 48 plus or minus 12 percent. The arterial pressure and blood viscosity decreased an average of 56 plus or minus 16 percent and 40 plus or minus 12 percent, respectively. The total peripheral resistance following the start of bypass was significantly below normal (p smaller than 0.005), but not when corrected for the change in viscosity. There was a strong correlation between the extent of pressure reduction and the viscosity decrease with hemodilution (p smaller than 0.01), although not all the changes in pressure could be accounted for in terms of viscosity. There was no apparent correlation between the extent of hypotension as measured by the area of the pressure-time curve below 50 mm. Hg and the presence or absence of postoperative cerebral or cardiac complications, in contrast to other recent reports.


Phagocytosis during general anesthesia in man. Latex particle phagocytosis and reduction of nitroblue tetrazolium by leukocytes was inhibited during general anesthesia when 18 patients were anesthetized with either halothane and nitrous oxide or nitrous oxide-thiopentalphils per 100 cells contained latex and 15.7 less monocytes per 100 cells contained latex. This represents a decrease of 17 and 28 percent, respectively, from preanesthetic values. The number of resting and latex-stimulated neutrophils reducing nitrobule tetrazolium was decreased 1.3/100 and 5.3/100, respectively, and the number of resting and stimulated monocytes reducing nitroblue tetrazolium was decreased 2.6/100 and 10.7/100, respectively. Phagocytosis was inhibited similarly by the two forms of anesthesia. The inhibition of phagocytosis during anesthesia in relatively healthy patients is minimal and its clinical significance has yet to be determined.


Urine norepinephrine excretion in patients undergoing mitral or aortic valve replacement with morphine anesthesia. Urine norepinehprine concentration and excretion were measured before, during and after mitral or aortic valve replacement in four groups of patients anesthetized with 1 to 3 mg/kg of morphine plus oxygen or 0.5 to 1.5 percent halothane and oxygen. All patients were similarly premedicated 90 minutes before operation. A bladder catheter was implanted at the time until anesthesia was begun. Urine was also collected during induction of anesthesia, from the first incision until bypass, during bypass, after bypass, and for 2 hours in the recovery room. Urine was analyzed for norepinphrine via the fluorometric method of Vikota. Urine output of patients undergoing the same prodecure were similar during all study periods irrespective of anesthetic. Mean preoperative urine norepinephrine concentrations and excretions of all groups were also similar. Urine norepinephrine concentrations and excretions in patients receiving halothane were unchanged until the postoperative period, when they became significantly increased. Patients given morphine anesthesia had marked increases in urine norepinephrine concentrations and excretions during induction, at all times intraoperatively, and postoperatively. These data suggest that morphine anesthesia increases norepinephrine blood levels in patients with valvular heart disease undergoing surgical correction.


Effects of different concentrations of corn oil, magnesium sulfate, and oat hulls in a porcine ulcerogenic ration. Two experiments involving 32 growing pigs were conducted to evaluate various concentrations of corn oil, magnesium sulfate, and oat hulls as ameliorants in a high cornstarch-low protein ulcerogenic ration. The pig were slaughtered at the end of the respective feeding periods, and stomachs were evaluated for issue lesions. During a 40-day feeding period, additions of corn oil and of MgSO4 in the rations did not seem to affect development of gastric lesions materially. After a 42-day feeding period, neither 5.0% nor 10.0% concentration of oat hulls in the ration prevented formation of ulcers. Total stomach contents, free gastric HCI, and pH of stomach contents were measured in pigs slaughtered after 3, 6, 9, and 12 hours of fasting. Gastric HCI (mEq/L) differed significantly among pigs fed various rations and increased significantly with length of fasting. The pH of stomach contents did not differ among pigs given various rations, but decreased with length of fasting. Thus, corn oil, MgSO4, or oat hulls did not seem to significantly alter the ulcerogenic character of the high cornstarch-low protein ration.


Prevalence of Dirofilaria immitis in a stray dog population in western New York. In a survey in 1973 undertaken to determine the prevalence of Dirofilaria immitis in 100 dogs from a public pound in western New York, 2 dogs were found to be positive. Microfilariae of Dipetalonema sp were not present. Hearts and pulmonary blood vessels were dissected for adult worms, and blood samples were examined for microfilariae, using a modified Knott technique. Although adult heartworms and microfilariae have been recovered from wild foxes, coyotes, and wolves in New York, this is the first report of heartworm in domesticated dogs in the state.


A new single-position supine approach to sciatic-femoral nerve block. To overcome the disadvantages of other methods, the authors have devised a procedure for below-the-knee operations, involving a new technic of sciatic-femoral nerve block. The technic has so far been successful without use of adjuvant agents in 90 percent of over 100 patients so handled.


Diagnostic value of serum creatine phosphokinase activity for the porcine malignant hyperthermia syndrome. Serum creatine phosphokinase (CPK) activity was measured in 10 German Landrace pigs from 11 to 28 weeks of age. A pronounced age dependence of serum enzyme activity was observed, peak activity occurring at 19 weeks of age. At the end of the growth period, when the pigs were challenged with halothane to detect the malignant hyperthermia syndrome, 3 pigs were found to be susceptible. Singificant increases in the serum enzyme levels in the susceptible pigs were observed only at 11 and 28 weeks of age. Serum enzyme levels measured during the rapid phase of growth could not be used to predict the malignant hyperthermia syndrome. Elevated serum CPK levels were also observed in two litters of Large White and Landrace x Large White pigs, breeds known to be stress-resistant. No pigs in these litters were susceptible to halothane, even though CPK levels were similar to those of the German Landrace pigs. The results indicate that serum CPK levels can be used as evidence of predisposition to the malignant hyperthermia syndrome but cannot be relied on as a single ultimate test.


Postoperative ventilatory failure in an obese, myopathic woman with periodic somnolence: A case report. An obese woman with periodic sommolence developed postoperative ventilatory problems following an uncomplicated cholecystectomy. A severe myopathic process involving the respiratory muscles was found at necropsy. A brother, who died a week later, had myotonic dystrophy.


Adequacy of main uterine vein and the ovarian artery in the local venoarterial pathway for uterine-induced luteolysis in ewes. The local uteroovarian pathway for uterine-induced luteolysis was studied in 34 ewes. Bilaterally ovulating ewes were used to study the role of the main uterine vein (uterine branch of uteroovarian vein) and unilaterally ovulating ewes were used to study the role of the ovarianartery. Surgical operation was done on day 7, 8, or 9 of diestrus and ewes were necropsied on day 20. In 4 bilaterally ovulating control ewes (unilaterally hysterectomized), mean weight of corpus luteum (CL) was less (P smaller than 0.05) on the side of the intact uterine horn (55 mg) than on the side of unilateral hysterectomy (548 mg). In 4 treated ewes (unilaterally hysterectomized and with surgical anastomosis of uterine vein from the intact side to the hysterectomized side), mean weight of CL on the hysterectomized side (153 mg) was less (P smaller than 0.05) than on the uterine-intact side (391 mg). The mean weight of CL on the hysterectomized side in the treated group was not significantly different from that of the uterine-intact side in the control group. Unilaterally ovulating ewes were randomized into 5 groups: (1) controls which were uterine intact, (2) controls in which a unilateral hysterectomy was done ipsilateral to CL, (3) unilateral hysterectomy done ipsilateral to CL and anastomosis of the ovarian branch of the ovarian artery on the uterine-intact side to the ovarian branch of the ovarian artery on the hysterectomized side, (4) controls in which unilateral hysterectomy was done contralateral to CL and the ovarian vascular pedicle on the uterine-intact side was isolated, and (5) unilateral hysterectomy done contralateral to CL, isolation of ovarian vascular pedicle on the uterine-intact side, and anastomosis of the ovarian branch of the ovarian artery on the unilaterally hysterectomized side to the ovarian branch of the ovarian artery on the uterine-intact side. Mean weight of CL was less (P smaller than 0.05) in groups 1 (56 mg), 3 (40 mg), and 4 (120 mg) than in groups 2 (408 mg) or 5 (473 mg). The mean weight of CL was not significantly different among groups 1, 3, and 4 or between groups 2 and 5. Results demonstrate that the local luteolytic effect of the uterus in ewes involves a venoarterial pathway and indicate that the main uterine vein is an adequate uterine outlet and the ovarian artery is an adequate ovarian inlet for the venoarterial pathway without the necessity of other possible concomitant local routes.


Quality-of-care appraisal in primary care: a quantitative method. A reproduceable method has been developed for measuring the quality of clinical care provided by physicians and nurse practitioners. The distinctive features of the method are the extended use of the tracer disease concepts, the evaluation of referrals, new procedures for probing the clinical operation of practices, a single blind design, emphasis on the use of the untouched medical record, the ability to compare results with measurements of concurrent outcome, and a relatively low cost. Three simultaneous approaches used in the method are described: surveillance of the management of indicator  conditions, evaluation of clinical use drugs, and the assessment of referral decisions. The three approaches gave consistently similar results about the relative performances of the practices compared and were in agreement with concurrent outcome studies. The method was successfully implemented in a health care experiment.


The pathogenesis of gluten-sensitive enteropathy. Gluten-sensitive enteropathy is characterized by flattening of intestinal villi and malabsorption caused by the toxic effect of gluten, a wheat protein. Gluten activates an endogenous mechanism of toxicity that may be the local mucosal immune system: local mucosal immunoglobulin and antigluten antibody production occur soon after gluten ingestion. Approximately 80% of patients with this disease possess HL-A8, a second segregant series antigen. This association also occurs in dermatitis herpetiformis, a disease with vesicular skin lesions and gluten-sensitive flattening of intestinal villi. The association suggests that the fundamental abnormality in enteropathy is a binding reaction involving gluten protein and a binding site on a cell surface, determined in part by the histocompatibility gene; this reaction then results in a local mucosal immune response to gluten. Alternatively, the fundamental abnormality may be the presence of an abnormal immune-response gene linked to the HL-A8 gene or acting in concert with it; this immune-response gene results in local mucosal production of antigluten antibody.


Antimicrobial activities of 81.723 hfu, a new pleuromutilin derivative. The new pleuromutilin derivative 81.723 hfu is extremely active against gram-positive organisms such as streptococci, staphylococci, and against mycoplasmas. A number of Shigella, Klebsiella, and Escherichia coli strains were also found to be quite susceptible to this new agent, whereas other gram-negative organisms like Pseudomonas aeruginosa, Proteus species, and Alcaligenes faecalis proved to be naturally resistant to 81.723 hfu. The new compound acts bacteriostatically. Bactericidal effects have been observed only at concentrations which are 100-fold higher than the minimal inhibitory concentrations. The new antibiotic is well tolerated in all animal species tested so far and has been successfully used in the treatment of experimental infections with gram-positive organisms and with mycoplasmas in mice and rats. Resistance against this new compound arose gradually in all microorganisms investigated. It is noteworthy that the rate at which resistance against 81.723 hfu emerged in mycoplasmas (Mycoplasma gallisepticum and Mycoplasma hyorhinis) was significantly slower than the corresponding rate at which resistance against tylosin tartrate appeared. Mycoplasma strains which became insensitive to 81.723 hfu were also resistant to tylosin tartrate, whereas mycoplasmas which developed resistance against tylosin tartrate, although less sensitive to 81.723 hfu than wild-type strains, were still eliminated by this drug. In a strain of Klebsiella pneumoniae, complete cross-resistance was observed between the pleuromutilin derivative on one hand and lincomycin and erythromycin on the other. Modest degrees of cross-resistance were also observed with chloramphenicol. However, it appears unlikely that the latter phenomenon is sufficiently pronounced to affect treatment with either antibiotic.


Glutamate dehydrogenase specific activity and cephalosporin C synthesis in the M8650 series of Cephalosporium acremonium mutants. Conditions for the accurate measure of glutamic dehydrogenase (GDH) from Cephalosporium acremonium were determined. K(m) values for alpha-ketoglutarate and ammonium ion were 7 and 15 mM, respectively. The half-saturation for reduced nicotinamide adenine dinucleotide phosphate was 5 muM. Reduced nicotinamide adenine dinucleotide did not serve as a cofactor for the enzyme. The specific activity of GDH was measured in six mutants of C. acremonium which varied in their ability to synthesize cephalosporin C. The mutants represented two separately derived lines, A and B. The four mutants in line B were characterized by a derepression of the GDH upon entry into stationary phase. The two mutants in line A were characterized by repressed levels of GDH during the same period. Both lines exhibited high GDH activity early in their fermentations, but activity decreased during the period of active cell growth. Cytochrome c concentrations followed the same pattern as total soluble intracellular protein. Line A mutants were low in cephalosporin C productivity and line B encompassed low, intermediate, and high productivity mutants. The relative frequency of yield improvements in line A and B indicate that the altered regulation pattern for GDH in line B may have removed a nitrogen limitation for cephalosporin C synthesis.


Dioxane treatment to improve microbial digestibility of cellulosic fibers. The hemicelluloses of grass seed straws can be extracted rather completely by acid detergent, but the treated residue (acid-detergent fiber) is normally not digested by rumen microorganisms. This residue can be made digestible if it is treated with acidic dioxane.


Factors influencing breast feeding.

Factors influencing breast feeding. A survey is reported of infant feeding practice at the time of discharge from a single maternity ward. It is shown that a change in the attitude of nursing staff increased the number of mothers breast feeding their infants and eliminated the practice of giving the infants formula feed complements. This change in attitude, however, did nothing to prevent the rapid decline in lactation after leaving hospital, such that 50% of mothers discharged breast feeding were no longer doing so by 2 months. The early introduction of solid feeds was common practice, 50% of mothers discharged breast feeding having introduced mixed feeds within 2 months. Successful lactation was found to be commoner in those mothers who had themselves been breast fed in infancy and in mothers from socioeconomic classes I and II. Successful lactation was not related to parity. The survey also shows the influential role of the health visitor, district nurse, and midwife upon mothers' decisions about infant feeding.


Infantile hypertrophic pyloric stenosis in Belfast, 1957-1969. Infants with hypertrophic pyloric stenosis born in Belfast during the 13 years 1957-1969 have been reviewed. Their distribution shows a bias towards higher social classes, breast feeding, and primogeniture. Obstetric factors and parental ages seem to be of no importance. More affected infants were born during winter months than would be expected. The overall incidence of infantile pyloric stenosis in this community has fallen during the period under review. Clinically, the patients started vomiting at a mean age of 22 days and it is recommended that the condition should not be called 'congenital'. The size of the tumour is mainly determined by the size of the patient, rather than by his age or duration of symptoms. Attention is drawn to the occurrence of haematemesis in 17-5% and melaena in 2-9% of infants. Jaundice occurred in 1-8% of patients in this series, and is attributed to the adverse effect of starvation on hepatic glucuronyl transferase activity. Other conditions noted in these patients included inguinal hernia, partial thoracic stomach, and phenylketonuria. Subsequent growth and development were in the anticipated range.


Unexpected postneonatal deaths (cot deaths) due to recognizable disease. At least a quarter of the babies dying as unexpected home 'cot deaths' do so from recognizable diseases. A study of the background of such children, compared with age and admission controls, suggests that the disease symptomatology in this group of children differs in no way from children who are successfully treated. The factors leading to the death of these children appear to lie in the general lack of ability of some parents to recognize the importance of symptoms, their negative attitude and inability to avail themselves of health services, and the amount of drive and persistence required to obtain general medical practice services in some areas. There is also the failure of some practitioners to recognize severely ill children.


Parathyroid hormone secretion during exchange transfusion. Plasma concentrations of calcium, phosphate, citrate, albumin, and parathyroid hormone (PTH) were measured during and after exchange transfusion of infants suffering from haemolytic disease using blood anticoagulated with acid-citrate and dextrose (ACD) or heparin. Pretransfusion plasma PTH and phosphate both correlated positively with postnatal age but not with each other. Transfusion with ACD blood caused a twelvefold rise in plasma citrate levels but no significant change in plasma calcium, phosphate, or PTH of the infant, despite the concentration of these substances being lower in the donor blood. The concentration of calcium, phosphate, and albumin was higher in heparinized than in ACD donor blood, and infants transfused with heparinized blood showed no change in the plasma concentration of any substance measured during transfusion. The addition of 50 mug glucagon to ACD donor blood had no effect on PTH secretion. 3 hours after transfusion there was a rise in the plasma PTH infants who had received ACD blood but not in those given heparinized blood. Transfusion with ACD blood caused a net loss of calcium, phosphate  and albumin from the infant, whereas transfusion with heparin blood did not. Both types of transfusion caused a net loss of PTH but this was significantly greater in those given ACD blood. These results show that transfusion with ACD blood results in increased secretion of PTH, probably due to the fall in ionized calcium concentration caused by the citrate load.


Blood coagulation status of small-for-dates and postmature infants. In a prospective study of blood coagulation status in small-for-dates and postmature infants there was often evidence of intravascular coagulation. Abnormal coagulation findings correlated with the degree of growth retardation and with the degree of postmaturity. Macroscopical placental infarction and neonatal polycythaemia were associated with coagulation abnormalities; asphyxia, however, was not. Intravascular coagulation may be an additional hazard to small-for-dates and postmature infants.


Physical growth in twins, postmature and small-for-dates children. From 16 955 livebirths, which occurred throughout the United Kingdom in April 1970 during the British Births Survey, three groups of children (multiple births, postmature, and small-for-dates) were selected together with a 10% random sample. This report is concerned with the height, weight, and head circimference measruements at 22 months of age. The babies that were heavier at birth showed diminished growth velocity and the small babies catch-up growth. At 22 months the pattern of distribution of weight was much closer to the normal than that of the babies' birthweights. Differences, however, did remain.


Neontal hypoglycaemia in infants of insulin-dependent diabetic mothers. Neonatal hypoglycaemia (blood glucose smaller than 20 mg/100 ml) occurred in the first 6 hours of life in 25 of 34 infants born to diabetic mothers receiving insulin. Despite severe hypoglycaemia (blood glucose smaller than 10 mg/100 ml) in 17, clinical features of hypoglycaemia were absent in all but 2. Hypoglycaemia was not related either to the level of plasma insulin in cord blood, determined as nonextracted immunoreactive insulin, or to the degree of control of maternal blood glucose during pregnancy. The frequent occurrence of severe neonatal hypoglycaemia in the infants born to diabetic mothers receiving insulin appears to be due rather to failure to maintain basal glucose homoeostasis after birth than to hyperinsulinism.


Vinyl chloride exposure in a controlled industrial environment. A long-term mortality experience in 594 employees. Vinyl chloride has been associated recently with findings of angiosarcoma in animals and man. The present study examines the mortality experience of individuals occupationally exposed to vinyl chloride and lesser amounts of vinylidene chloride and other compounds. Employees were grouped into four exposure categories according to the highest levels of vinyl chloride exposure experienced for at least one month. Although no angiosarcomas were found and there were no deaths due to any liver malignancy, the observed malignancy deaths exceeded the expected among workers in the high-exposure category. Fewer than expected malignancy deaths were observed in the remaining exposure catergories.


The diagnostic accuracy of palpation and fine-needle biopsy and an evaluation of their combined use in the diagnosis of breast lesions: report on a prospective study in 1244 women with symptoms. In 1244 women with breast symptoms an evaluation by means of palpation was made with respect to diagnosis of malignancy according to a four-grade scale ranging from "definite cancer" to "no cancer." Aspiration biopsy and cytologic examination were then performed in 984 breast lesions. The diagnosis from the cytologic evaluation ranged from benign, through three grades of atypia (slight, moderate, grave) to cancer. A histologic diagnosis was made in 411 cases and in 28% it was cancer. Cancer was found in 92.5% of the patients with a palpatory diagnosis of "definite cancer," and in 50% of those with a palpatory diagnosis of "strong suspicion of cancer." In all patients in whom cancer was diagnosed cytologically, the same diagnosis was made at histology, while 87.5% of those with grave atypia at the cytologic examination were diagnosed histologically as having cancer. A false negative cytologic diagnosis was made in 4% of the cancer cases. With a combination of palpation and cytology, 91% of the cancer cases fell within the groups "definite cancer," "strong suspicion of cancer" (palpation)/"cancer," "grave atypia" (cytology). No patients with cancer were evaluated as "no cancer" (palpation)/"no atypia" (cytology). In this group of 697 patients, however, one cancer was discovered after 7 months. The investigation showed that a thorough palpatory evaluation is a prerequisite for a good result of aspiration biopsy, in particular to meet the risk of a false negative cytologic diagnosis. The cytologic examination revealed cancer in 6 and 12 cases, respectively, when palpation gave no or some suspicion of cancer, and in many cases it was able to eliminate malignancy suspected on palpation. Cytologic atypia indicated cancer in a relatively high per cent, but was also noted in many cases found to be benign histologically. The possibilities of reducing the number of "unnecessary" surgical biopsies by using a combination of palpation and cytology is discussed.


Brucellosis outbreak attributed to ingestion of umpasteurized goat cheese. In six cases  of Brucella melitensis infections the common etiologic factor was the ingestion of raw goat cheese. Presenting complaints were variable, but a constant feature was persistent fevers that frequently were present for months. Evidence of liver cell dysfunction was present in each case, and in two, biopsy disclosed diffuse hepatitis with focal necrosis. The diagnoses were initially based on high titers of Brucella agglutinins and later confirmed by positive blood cultures. A good clinical response was induced by antibiotic therapy ( tetracycline plus streptomycin sulfate), but in two cases a Jarish-Herxheimer reaction occurred during the first 24 hours of treatment.


Influence of concanavalin a on autolysis of gametes from Chlamydomonas reinhardii. The phytohemagglutinin concanavalin A inhibited zygote formation of Chlamydomonas reinhardii. 15--50 mug lectin/ml not only interfered with the mating reaction, but also with cell wall lysis of gametes and zoospores in a crude autolysin preparation gained from copulating gametes. Further, the structure of cell walls shed into the medium after autolysis in the course of the mating reaction and after lysis "from without" in the crude autolysin preparation was stabilized by Con A. Therefore, it must be assumed that the lectin inhibited zygote formation of C. reinhardii by interfering with autolysis of the cell walls of the gametes. Though Con A inhibited the lytic processes of C. reinhardii, an activation of the autolytic system in theta gametes by the lectin was found to compete with its inhibitory reaction. Con A induced autolysis of theta gametes was dependent on adherence of the cells by their flagella to the surface of the culture vessel or the liquid medium and did not occur in cultures stirred by rotation. The interferences of Con A with the autolytic serum of C. rienhardii were inhibited by methyl-alpha-D-mannopyrano-side and to a lesser degree by glucose, indicating that the carbohydrate binding sites of the lectin were involved in its reactions with the cells.


Peripheral blood and myelogram in white rats chronically poisoned with ethanol and receiving essential phospholipids. The influence of chronic administration of ethyl alcohol in white rats (15% solution instead of water, ad libitum, during 8 months) and essential phospholipids on the peripheral blood picture and myelogram was studied. Hemoglobin, erythrocyte counts and leukocytes dropped uniformly in the group of animals poisoned with alcohol. Stimultaneous administration of essential phospholipids exerted a protective effect with respect to the fall in hemoglobin and erythrocyte counts.


Replication of eastern equine encephalitis viruses (New Jersey and Louisiana strains and the Ets-4 mutant) in rabbit kidney cells. Virus yield, viral RNA synthesis, viral protein synthesis, cytopathology, and virus-induced shutoff of host protein synthesis were examined in rabbit kidney (RK) cells infected with eastern equine encephalitis (EEE) viruses. The New Jersey (NJ) strain replicated most rapidly, and exhibited a distinct patter in the synthesis of structural and non-structural viral proteins. The Louisiana (LA) strain and the Ets-4 mutant appeared to be similar in RK celles and chick embryo fibroblasts. Although the viral peptides comprising the viral RNA polymerase could not be conclusively identified, two non-structural proteins of molecular weights 105,000 and 85,000 were temporally associated with increasing rates of polymerase activity in NJ-infected RK cells and were found in increased amounts in Ets-4-infected cells compared to La-infected cells.


A lipid inhibitor of dengue virus in human colostrum and milk; with a note on the absence of anti-dengue secretory antibody. Neutralizing activity against dengue virus types 1--4 was observed in milk samples from 5 non-immune and 29 dengue immune women. Anti-dengue activity in milk and colostrum was found only in the lipid component. The inhibitory activity is directed against the virus and not cell surfaces. When immunoglobulin types IgM,IgA, IgG were isolated from colostrum from dengue immune women, no antibody activity was found. Anti-dengue activity in human milk did not decrease over a period of ten months after delivery.


Serologic evidence for etiologic role of Akabane virus in epizootic abortion-arthrogryposis-hydranencephaly in cattle in Japan, 1972-1974. In the outbreak of abortions, premature births, stillbirths and congenital arthrogryposis-hydranencephaly (AH) syndrome in Japan during the summer through winter of 1972-73 and 1973-74, precolostral sera from calves with congenital AH syndrome and normal calves were tested for neutralizing antibodies against some arboviruses, i.e. Akabane, Aino, Getah and Japanese encephalitis (JE) viruses. The incidence of antibody for Akabane virus was very high in calves with AH syndrome (49/59 or 83 per cent) as compared with normal calves (3/11 or 27 per cent), indicating an intimate correlation between the AH syndrome and precolostral anti-Akabane antibody. Three stillborn fetuses also had anti-Akabane antibody. On the other hand, no precolostral serum antibody for the other viruses was detected in any of the calves tested. The mothers of these calves, normal and with AH syndrome, had anti-Akabane antibody in high percentages (44/52 or 85 per cent and 7/8 or 88 per cent), whereas a few of the mothers had antibodies for the other viruses. Serological surveys indicate a wide dissemination of Akabane virus in epizootic areas during the summer months of 1972 and 1973. Thus, 8 groups of cattle in epizootic areas showed high rates of seroconversion for Akabane virus during the 1972 or 1973 summer. Very high incidences of Akabane antibody were shown among cattle in epizootic areas but extremely low incidences in near-by non-epizootic areas. The geographic distribution of anti-Akabane antibody among cattle throughout the country in the 1973 spring generally agrees with the pattern of case distribution in the 1972--73 outbreak. All these findings strongly suggest that Akabane virus is the etiological agent of the outbreaks. Further studies are needed, particularly isolation of the virus, demonstration of infection with the virus in lesions by immunofluroescence and production of intrauterine infection by experimental infection of pregnant cows.


Rubeosis in Fuchs heterochromic iridocyclitis. A patient had Fuchs heterochromic iridocyclitis of approximately 17 years' duration. Histologically, keratic precipitates, iris atrophy, rubeosis iridis, discontinuous rubeosis of the anterior chamber angle, a chronic nongranulomatous iridocyclitis, and trabeculitis were noted. The cause of the glaucoma probably is a combination of rubeosis of the anterior chamber angle and trabeculitis.


Laser iridotomy for aphakic pupillary block. In six patients with aphakic pupillary block, argon laser produced full-thickness iridotomies that succeeded in restoring anterior chamber and intraocular pressure to normal. In one case, it was shown that location of iridotomy is important in determining whether or not iridotomy will succeed in relieving aphakic pupillary block. In two cases, pupillary block had resulted in shallowing of the anterior chamber without pressure elevation. Cases reported here show that continuous laser energy, such as from an argon laser, can produce a full-thickness opening, in contrast to the short duration pulse of the ruby, which will destroy only pigmented structures without producing a full-thickness iris opening. This need to produce some degree of spread of damage to unpigmented tissue elements in order to achieve and iris opening is a problem in phakic patients where injury to the lens must be avoided.


Iris photocoagulation therapy of aphakic pupillary block. Five patients who underwent uneventful cataract extraction developed flat anterior chamber (AC) secondary to pupillary block, 2 1/2 to 4 weeks postoperatively. None had a wound leak. After drug therapy had failed to restore the AC, iris photocoagulation was performed. Four eyes were treated with a xenon-arc photocoagulator, and one with both a portable xenon-arc unit and an argon laser. The AC deepened immediately and permanently in all cases. No complications occurred, and visual acuity was good. Iris photocoagulation with a xenon-arc unit is a valid alternative to intraocular surgery in therapy for flat AC that is caused by aphakic pupillary block in patients in whom medicinal therapy fails and no wound leak is present.


Lipid metabolism in the cow during starvation-induced ketosis. 1. Concentrations and compositions of liver, serum and milk lipids of cows were measured during 6 days' starvation and serum lipids during 60 days' re-feeding. 2. The concentration of free fatty acid in serum increased fivefold during starvation. 3. The content of total lipid in liver (g/100g of liver dry matter) doubled owing to a 20-fold increase in triglyceride, an eightfold increase in cholesterol ester, a three fold increase in free fatty acid and a 20% increase in cholesterol. There were no changes in the content or composition of liver phospholipids. 4. Starvation lowered the concentrations of total lipid, phospholipid and cholesterol ester of dextran sulphate-precipitable serum lipoproteins. Total lipid and cholesterol ester concentrations in lipoproteins of d greater than 1.055 and in lipoproteins not precipitable by dextran sulphate decreased from day 4 of the starvation period and during the first 20 days' re-feeding. 5. During starvation there were decreases in percentages of stearic acid and increases in oleic acid in serum free fatty acids and triglycerides and in liver neutral lipid. 6. Throughout starvation total milk lipid yield decreased, yields and percentages of C4-14 fatty acids decreased and percentages of C18 fatty acids increased. 7. It is suggested that accumulation of triglyceride in liver may be caused by increased uptake of plasma free fatty acids without corresponding increase in lipoprotein secretion.


Chemcial anatomy of the nerve membrane.

Chemcial anatomy of the nerve membrane. The molecular mechanisms involved in the generation and conduction of nerve impulses are still matters for conjecture. The molecular models which have been developed to explain the structure of nerve membrane are reviewed. The chemical composition of the axolemma and how this compares with other membranes is discussed. Modern spectroscopic techniques allow the measurement of non-electrical dynamic changes that occur during nerve conduction. These indicate that the chemical components of nerve membrane are in a very fluid state. Detailed knowledge of the kinds of molecules involved is necessary to understand properly both nerve conduction and nerve block.


Primary cell culture for biochemical studies of human keratinocytes. A method for production of very large numbers of cells without the necessity of subculturing techniques. Starting with large numbers of small split-thickness explants of human skin, it is possible to grow ample numbers of keratinocytes in vitro as primary cell cultures to permit biochemical studies without the need of subculturing techniques.


Course in basic sexology for medical students. The present article describes the pilot course in basic sexology for medical students. The duration of the course was 5 days (35 hours). The themes-sex and gender, sexual physiology, contraception, sexual inadequacy, sexual deviations, and sexual counselling-were approached from many different angles. The teaching procedure comprised lectures, group work, and group discussions, internal television, films, plenary discussions, and debates. The course was evaluated by means of the sex knowledge and attitude test (SKAT) given before and after the course. Furthermore an evaluation was given by the students, by a professional teacher, and by a paramedical evaluator at the end of the course. The results showed significant changes in attitudes and knowledge.


Metabolic responses following isoproterenol-induced myocardial infarction in arteriosclerotic breeder vs. non-arteriosclerotic virgin and ovariectomized female rats. Intact and ovariectomized, non-arteriosclerotic female rats and arteriosclerotic, breeder female rats were subjected to myocardial infarction by the administration of 2 subcutaneous injections, 24 hr apart, of the beta-adrenergic stimulator, isoproterenol. The animals were sacrificed at regular hourly intervals following each injection and then on days 4, 6, 8, 10, 12, and 16 thereafter. Measurement of serum insulin and free fatty acids (FFA) demonstrated a blunted response in these metabolic parameters in the case of the ovariectomized virgin rats. The non-arteriosclerotic, intact virgin rats exhibited dynamic changes in serum insulin, glucose, FFA and corticosterone (Cmpd. B) following both isoproterenol injections, whereas the arteriosclerotic breeder rats manifested elevated levels of these parameters on Day 1, but did not display any increases in serum levels, except for FFA, following the second isoproterenol injection on Day 2. In spite of the hormonal alterations brought about by the gonadectomy and their differing metabolic response, the ovariectomized females did not have a mortality rate significantly different from the intact females. Thus, the presence or absence of ovarian estrogens seemingly does not affect the progress of an isoproterenol-induced myocardial infarction.


Plasma concentration of gonadotrophins, oestrogens and progesterone in thyrotoxic women. Plasma levels of luteinizing hormone (LH), follicle stimulating hormone (FSH), oestrogen and progesterone were measured daily in 15 thyrotoxic women in the reproductive age for 28 to 30 consecutive days before commencement of therapy and for a similar period following restoration of the euthyroid state.  Five of these patients had secondary amenorrhoea whilst the other 10 had hypomenorrhoea.  Twelve healthy volunteer eythyroid women of similar age and parity, with no history of menstrual abnormality, served as controls.  The levels of both gonadotrophins (LH and FSH) and oestrogen were found to be significantly raised in all thyrotoxic patients in comparison with levels in the euthyroid female controls (P less than 0.001).  "Mid cycle" LH and FSH peaks were present in thyrotoxic patients who were still menstruating whilst they were absent in those who developed amenorrhoea.  Plasma oestrogen concentration rose to a peak on the day before the surge of LH (and FSH) in all euthyroid frmale controls as well as in thyrotoxic patients who were still menstruating.  In contrast, oestrogen peaks occurred without ensuing LH (and FSH) peaks in thyrotoxic patients who had developed amenorrhoea, suggesting a failure of the positive feed-back mechanism.  The levels and patterns of plasma progesterone in thyrotoxic subjects who were still menstruating were similar to those of euthyroid controls.  However, no significant circulating plasma progesterone was detected in thyrotoxic subjects who were amenorrhoeic.  The levels and patterns of all of these hormones (gonadotrophins, oestrogen and progesterone) became normal after restoration of euthyroid state and normal menstruation.


Kinetic analysis of the mechanism of insulin degradation by glutathione-insulin transhydrogenase (thiol: protein-disulfide oxidoreductase). Kinetic studies have been made with glutathione-insulin transhydrogenase, an enzyme which degrades insulin by promoting cleavage of its disulfide bonds via sulfhydryl-disulfide interchange. The degradation of 125I-labeled insulin by enzyme purified from beef pancreas was studied with various thiol-containing compounds as cosubstrates. The apparent Km for insulin was found to be a function of the type and concentration of thiol; values obtained were in the range from 1 to 40 muM. Lineweaver-Burk plots for insulin as varied substrate were linear, whereas those for the thiol substrates were nonlinears: the plots for low molecular weight monothiols (GSH and mercaptoethanol) were parabolic; those for low molecular weight dithiols (dithiothreitol, dihydrolipoic acid, and 2,3-dimercaptopropanol) were apparently linear modified by substrate inhibition; and the plots for protein polythiols (reduced insulin A and B chains and reduced ribonuclease) were parabolic with superposed substrate inhibition. The nonparallel nature of the reciprocal plots for all substrates shows that the enzyme does not follow a ping-pong mechanism. Product inhibition studies were performed with GSH as thiol substrate. Oxidized glutathione was found to be a linear competitive inhibitor vs. both GSH and insulin. The S-sulfonated derivative of insulin A chain was also linearly competitive vs. both substrates. Inhibition by S-sulfonated B chain was competitive vs. insulin; the data eliminated the possibility that this derivative was uncompetitive vs. GSH. Experiments with the cysteic acid derivatives of insulin A and B chains similarly excluded the possibility that these were uncompetitive vs. either substrate. These inhibition studies indicate that the enzyme probably follows a randdom mechanism.


Interaction of insulin analogs, glucagon, growth hormone, vasopressin, oxytocin, and scrambled forms of ribonuclease and lysozyme with glytathione-insulin transhydrogenase (thiol: protein-disulfide oxidoreductase): dependence upon conformation. Interactions of several proteins with glutathione-insulin transhydrogenase (GIT) have been investigated by determining their ability to inhibit degradation of 125I-labeled insulin catalyzed by GIT. The inhibition by every insulin analog (des-Asn-des-Ala-pork insulin, desoctapeptide-pork insulin, des-Ala-pork insulin, pork insulin, proinsulin, and guinea pig insulin) was competitive vs. competitive vs. insulin indicating that they function as alternate substrates. The insulin analogs with the least hormonal activity showed the highest potency as inhigitors of insulin degradation. Whereas native ribonuclease and lysozyme showed little or no inhibition, their scrambled forms (i.e. reduced and randomly reoxidized) showed competitive inhibition with a potency greater than that of insulin. These results suggest that the conformation of the substrate or inhibitor is probably the major factor in determining the specificity for (or binding to) the enzyme. Studies withother peptide hormones showed competitive inhibition with vasopressin and oxytocin and noncompetitive inhibition with glycagon. The inhibition with growth hormone could be either competitive or noncompetitive. The inhibition by glucagon and growth hormone (physiologic antagonists of insulin) could serve as a control mechanism to modulate the activity of enzyme. The following showed very little or no inhibition; the native and scrambled form of pepsinogen, trypsin inhibitor of beef pancreas and of lima bean, C-peptide of pork proinsulin, and heptapeptide (B23-B29) of insulin.


Application of pulsed-gradient Fourier transform nuclear magnetic resonance to the study of self-diffusion of phospholipid vesicles. A pulsed-gradient Fourier transform nuclear magnetic resonance (NMR) technique was appplied to the study of diffusion of phospholipid vesicles. The diffusion coefficient of dimyristoyllecithin vesicles (DML) in a D2O-phospahte buffer at 37 degrees is D = 1.9 TIMES 10(-6) cm2/sec. In a solution made viscous by DNA addition, the diffusion coefficient of DML vesicles was 3.5 times 10(-7) cm2/sec. These values compare favorably with the diffusion rate for liposomes as determined by ultracentrifugation and by Stokes law calculation. The data suggest that DML diffusion is controlled primarily by whole liposome migration as opposed to movement of individual molecules within the liposome, liposome rotation, or fast exchange between lecithin molecules in solution and in vesicles.


Cell contact-dependent ganglioside changes in mouse 3T3 gibroblasts and a suppressed sialidase activity on cell contact. Certain enzyme activities for synthesis and degradation of gangliosides and the chemical quantity and incorporation of radioactivity from [14C] galactose into gangliosides have been studied in 3T3 cells and their transformed counterparts at various cell population densities. The chemical quantity of and the incorporation of radioactivity into GD1a ganglioside increased at the early stage of cell contact ("contact response" of ganglisoide), whereas response was not detectable in transformed 3T3 cells at any stage of cell contact. These phenomena were reproduced in five separate qualitative analyses and two quantitative determinations of gangliosides. As the basis of these phenomena, a membrane-bound sialidase activity which acted on gangliosides was suppressed in 3T3 cells at the "touching" stage of cell-to-cell contact. Transformed cells did not display the change of sialidase activity at any stage of cell contact.


The complete amino acid sequence of ubiquitin, an adenylate cyclase stimulating polypeptide probably universal in living cells. The complete amino acid sequence was determined  for bovine ubiquitin, and adenylate cyclase stimulating polypeptide, which is probably represented universally in living cells. Ubiquitin has a molecular weight of 8451 and consists of a single polypeptide chain containing 74 amino acid residues. It contains four arginine residues but no cysteine or trytophan residues. The first 61 amino acid residues were obtained by automated Edman degradations. Tryptic digestion of maleated ubiquitin yielded four peptide fragments that were resolved by molecular sieve chromatography and coded in order of decreasing chain length (MT-1, MT-2, MT-3, and MT-4). The automated sequenator determinations on native ubiquintin provided overlapping sequence data for three of these fragments that gave an order of MT-1, MT-3, and then MT-2; Peptide MT-4, a dipeptide, was therefore assigned to the C terminus, and the placement of peptide MT-2 was corroborated by analysis of data from carboxypeptidase digestions of maleated ubiquitin. Peptide MT-2 was domaleated and sequenced by manual Edman degradations through a single lysine residue. It was cleaved at this residue with trypsin, and the two resultant peptides were separated by ion-exchange chromatography. Manual sequencing of the C-terminal demaleated tryptic peptide of MT-2 completed the sequence of MT-2 and that of native ubiquitin. The sequence of ubiquitin was further confirmed and supported by amino acid and parital sequence anlysis of fragments obtained by digestion of maleated ubiquitin with chymotrypsin or staphylococcal protease.


1-Halo analogs of dihydroxyacetone 3-phosphate. The effects of the fluoro analog on cytosolic glycerol-3-phosphate dehydrogenase and triosephosphate isomerase. Fluoro-o-hydorxyacetone phosphate (fluoroacetol phosphate) has been prepared by oxidation of 1-fluoro-3-chloro-2-propanol to 1-fluoro-3-chloroacetone, phosphorylation with silver dibenzylphosphate, and the intermediate isolation of 1-fluoro-3-hydroxyacetone phosphate dibenzyl ester, followed by catalytic hydrogenation and preparation of the stable monosodium salt. The chloro analog as the pure, stable monosodium salt has been prepared by a similar route from 1,3-dichloroacetone. 1-Fluoro-3-hydroxyacetone-P is substrate for cytosolic NAD+-linked glycerol-3-P dehydrogenese (EC 1.1.1.8) from rabbit skeletal muscle with an apparent Km of 50 mM under conditions in which dihydroxyacetone-P exhibits an apparent Km of 0.15 mM. Under these conditions the fluoro analog is 85% hydrated wheras dihydroxyacetone-P has been shown by others to be 44% hydrated. The turnover numbers are 49,000 molecules of NADH oxidized per minute per molecule of enzyme at 25 degrees with the fluoro analog as substrate, and 60,000 with dihydrocyacetone-P as substrate. The product of the reduction of the fluoro analog has been identified as 1-fluorodeoxyglycerol-3-P. 1-Fluoro-3-hydroxyacetone-P is comparatively weak irreversible inhibitor at 4 degrees of rabbit muscle triosephosphate isomerase (EC 5.3.1.1) with second-order rate constant of 2.6 M minus 1 sec minus 1. Inhibition by pyrazole in vivo of alcohol dehydrogenese catalyzed oxidation of 1-fluorodeoxyglecerol-3-P indicates in mice the reduction of 1-fluoro-3-hydroxyacetone-P to -l-1-fluorodexoxyglycerol-3-P  is not significant metabolic route, or that an alternative route exists when the alcohol dehydrogenase dependent pathway is inhibited.


Bicarbonate stabilization of ribulose 1,5-diphosphate carboxylase. The carboxylase and oxygenase activities of purified soybean ribulose 1,5-di-P carboxylase (EC4.1.1.39) were unstable when reactions were initiated with enzyme. Time courses of carboxylase and oxygenase activities were curvilinear, approximating hyperbolas. Double reciprocal plots of amount of CO2 incorporated and P-glycolate produced vs. time were constructed to determine a constant representing the half-time of initial enzyme activity, K. K increased with increasing bicarbonate concentration but was independent of O2 tensions between 0.21 and 5 atm. When time courses of carboxylase and oxygenase activities were determined simultaneously, K was identical for both activities. Linear time courses were obtained py preincubation of the enzyme for 10 min in the absence of bicarbonate or by adding 46 mM MgCl2 to the reaction mixture. The observed bicarbonate-dependent decline in ribulose 1,5-di-P carboxylase activity with time is the probable cause for the anomalously high Km(CO2) values previously reported for this enzyme. In the experiments reported here, the apparent Km(CO2) at pH 8.5 increased from 6 muM CO2 at zero time to 78 muM CO2 at 10 min. The corresponding bicarbonate Km values ar 1;3 and 17 mM, respectively, The interaction between bicarbonate and enzyme may be important in the light activation of photosynthetic CO2 fixation in vivo.


The control of prothrombin conversion. Kinetic control by mechanisms inherent in two activation pathways. It is known that the activation of prothrombin to thrombin can proceed via two pathways: one initiated by the prothrombin-converting complex (factor Xa, factor V, phospholipid, and CA2+ ions) and the other initiated by the product, thrombin. A kinetic study has shown that the pathways do not proceed with equal ease under all conditions. At high levels of the converting complex, both go to completion: some prothrombin is always cleaved by thrombin, but the resulting intermediate is then activated to give quantitative conversion to thrombin. At slower rates of activation, the product-initiated pathway occurs to a relatively greater extent. Moreover, the intermediate then is not cleaved further but accumulates, so that the generation of thrombin is curtailed. The reason the intermediate is productive only at higher levels of activator may be partly that it is a poorer substrate for the converting complex than prothrombin. More importantly, the activity of the complex is also modulated by thrombin, which rapidly destroys the activity of factor V and factor Xa in a feedback reaction. These concerted controls ensure that prothrombin activation damps itself. Thus thrombin production occurs as a burst, the size of which is regulated by the amounts of factor Xa and factor V initially available.


The conformation of proteins in chromatin. A circular dichroism study below 250 nm. This paper is an investigation of the circular dichroism (CD) spectra of DNA and protein in chromatin. The circular dichroism (CD) of chromatin below 250 nm is due to DNA and protein peptide chromophores. The spectrum in this region is resolved into contributions from salt-extractable proteins (histone and non-histone proteins extractable with sodium chloride), residual non-histone proteins (not extractable with 3 M sodium chloride), and DNA. Below 250 nm, DNA in chromatin has the same CD spectrum as DNA free in solution, in contrast to the CD of DNA above 250 nm (Hjelm, R. and Huang, R. C., (1974), Biochemistry 13, 5275). Histones and salt-extractable non-histone proteins in chromatin are seen to have an average CD like those observed for globular proteins. The average CD of the residual non-histone proteins is consistent with a population of proteins with more extended conformation. The CD of each of these components is found to be the same in chromatins isolated from tissues having different nuclear synthetic activities: chick embryo brain, pig cerebellum, myeloma K41, calf thymus, and chicken erythrocyte.


Growth hormone covalently bound to sepharose or glass. Analysis of ligand release rates and characterization of soluble radiolabeled products. Purified boveine growth hormone labeled enzymatically with iodine-125 was covalently coupled to cyanogen bromide activated Sepharose 4B gel and to diazotized zirconia-clad glass beads. Under the conditions employed, an average of 0.8 and 7.3 mg of hormone were bound per ml of Sepharose and glass, respectively. When the conjugates were incubated in Krebs-Ringer bicarbonate buffer (pH 7.4), three separate radioactive species were detected in the incubation supernatant by chromatography on Sephadex G-75. The elution volumes of two of the species were identical with those of 125-I-labeled growth hormone and Na-125I controls, while the third component eluted as a moleucle of intermediate size. The rate of release of each species from the solid matrix was linear with time over 4 days and increased with temperature from 4 to 37 degrees. Although significantly less growth hormone was released from glass (0.14%/day) than from Sepharose (0.40%/day) at 37 degrees, active hormone in amounts sufficient to be detectable in a biological assay was nevertheless liberated from the former after as little as 4 hr of incubation. By contrast, the rate of release of 125-Iminus- and the intermediate-size compound from glass was significantly greater than from Sepharose, suggesting that protein bound to glass supports is more susceptible to degradation from exposure to ionizing radiation.


Changes in template activity and structure of nuclei from WI-38 cells in the prereplicative phase. Quiescent confluent monolayers of WI-38 fibroblasts were stimulated to proliferate by either adding 10% fetal calf serum or by trypsinization and replating at lower density. The length of the prereplicative phase was 12 hr after serum stimulation and 18 hr after trypsinization and replating at lower density. Nuclei were isolated from WI-38 cells at different time intervals after either type of stimulation and their template activity, circular dichroism spectra, and ability to bind ethidium bromide were investigated. All these parameters were similarly increased after either type of stimulation. However, these changes, like the onset of DNA synthesis, were delayed 6 hr in cells trypsinized and replated at lower density. While there were no detectable changes in nuclear protein content after serum stimulation, at least 40% of nuclear protein, mostly nonhistone chromosomal proteins, were lost after trypsinization. The amount of nuclear proteins returned to prestimulation levels only 6-8 hr after replating. These data seem to suggest that nonhistone chromosomal proteins lost by trypsinization are essential for the entrance of WI-38 cells into the "prereplicative phase".


DNA chain termination by 2',3'-dideoxythymidine in replicating mammalian cells. The thymidine analog, 2,3-dideoxythymidine (ddT), is rapidly phosphorylated and incorporated terminally at 3-ends of growing DNA chains in replicating mammalian cells. Following some initial loss of ddT incorporated into DNA chains, the major portion is retained for periods equivalent to more than two normal cell generations. Some ddT appears at the termini of oligonucleotides, a portion of which have chromatographic properties suggesting internally complementary sequences. While these oligonucleotides may include degraduation fragments, it is possible that some represent replication initiation sequences.


Structural studies on bovine prothrombin. Isolation and partial characterization of the Ca2+ binding and carbohydrate containing peptides of the N-terminus region. Three peptides, one of which binds Ca2-+ (calcium binding fragment, CBF) but contains no carbohydrates and two of which bind no Ca2-+ but contain carbohydrates, have been isolated from the N-terminus region of bovine prothrombin. The preparation of these peptides involved (a) thrombin cleavage of prothrombin to intermediate 1 (thrombinogenic) and fragment 1 (nonthrombinogenic), (b) tryptic attack on fragment 1, and (c) separation of the CBF from the latter reaction by addition of a phosphatidylcholine-phosphatidylserine dispersion in the presence of Ca-2+. Further study on the non-calcium-binding peptides from the tryptic digest of fragment 1 revealed the presence of two low molecular weight glycopeptides, GP-1 and GP-2. A detailed examination of the chemical characteristics of CBF provided some insight into this unusual peptide. Whereas fragment 1, as well as prothrombin, exhibited two classes of Ca-2+ binding sites (one of high affinity, 3-4 mol/mol of peptide and the other of low affinity, 10-12 mol/mol of peptide), CBF bound only 3-4 mol of Ca-2+/mol of peptide. This indicated the presence of only the high affinity sites of the parent molecule. CBF contained an unusually high level of glutamic acid (approximately 30% of the total amino acids as determined in an acid hydrolysate) and had an N-terminal glycine. Most likely these glutamyl residues were present originally as the gamma-carboxyglutamyl residue as proposed by Stenflo et al. (Stenflo, J., Ferlung, P., Egan, W., and Roepstorff, P. (1974), Proc. Natl. Acad. Sci. U.S-A 71, 2730). The CBF contained no detectable carbohydrate. Its molecular weight varied inexplicably according to the procedure used and gave the following values; 8500, by gel filtration; 5200, by 6 M guanidine-HCl gel chromatography; 3490, by analytical ultracentrifugation. The glycopeptides, GP-1 and GP-2, were distinguished from each other by differences in their behavior on ion exchange chromatography and in their amino acid composition, and from CBF by their inability to bind calcium under any conditions. On the other hand, GP-1 and GP-2 had nearly identical levels of carbohydrate, 45.1 and 48.0 wt %, and possessed essentially the same percent distribution of carbohydrates: sialic acid, 16.5 plus or minus 0.5; mannose, 10.3 plus or minus 0.4; glucosamine, 11.2 plus or minus 0.1; galactose, 7.9 plus or minus 0.3. Their molecular weights were as follows: GP-1, 70000, by gel filtration; 6500, by 6 M guanidine-HCl gel chromatography; 4600, by ultracentrifugation; GP-2, 6500 by gel filtration; 6900, by 6 M guanidine-HCl gel chromatography; 1960, by analytical ultracentifugation. Though there are some obvious variations depending on method, this could be attributable to a probable error in v measurement on these carbohydrate containing peptides. The significance of these findings as they relate to prothrombin to thrombin conversion is discussed.


Dynamic equilibrium in histone assembly: self-assembly of single histones and histone pairs. The assembly of acid-extracted, purified F2a1, F3, F2a2, and F2b histones and their six possible pairwise combination into organized structures has been studied by: (1) sedimentation velocity, (2) sedimentation equilibrium, (3) electrophoresis in polyacrylamide gels containing sodium dodecyl sulfate after cross-linking the protein solution with dimethyl suberimidate, and (4) electron microscopy. Each of the purified histone fractions can renature and assemble into high molecular weight organized structures. This assembly is dependent on the ionic strength, protein concentration, and temperature of the solutions. The four histones studied assemble into structures of similar dimensions and shape. In each case the first structure observed is a bent rod with a diameter of 22 A. Conditions which favor assembly lead to formation of fibers with diameters of about 44 A. The conditions which lead to assembly into organized structures are similar for the arginine-rich histones, F2a1 and F3. Higher ionic strength is required for the assembly of the lysine-rich histones, F2a2 and F2b. Certain pairs of histones interact. Strong interactions among pairs of histones interfere with the self-assembly of single histones into large structures. Howver, increase in protein concentration or ionic stregth leads to formation of large molecular structures even in solutions of pairs of strongly interacting histones. These structures are similar to those obtained with single histones. The results suggest that aggregation and complexing of histones represent a reversible, ordered process of assembly. The various assembled forms are in a dynamic equilibrium. The final assembled form, which is similar in all cases, is dependent on the environmental conditions to which the histones are exposed. It is suggested that each of the assembled histone structures, regardless whether it is composed of a single histone or a pair of histones, can serve as a core around which the DNA can be wrapped.


Structural studies on phophatidylcholine-cholesterol mixed vesicles. The homogeneous, single-walled phosphatidylcholine-cholesterol mixed vesicles were prepared by ultrasonic irradiation of egg phosphatidylcholine in the presence of various amounts of cholesterol in solution at 4 degrees under a nitrogen atmosphere followed by molecular sieve chromatography on a Sepharose 4B column. Physicochemical studies performed on these systems invluding sedimentation velocity, diffusion, partial specific volume, intrinsic viscosity, and trapped volume measurements allowed estimation of the weight-average vesicle weight, the vesicle shape, and bilayer membrane thickness of the binary mixture of phosphatidylcholine and cholesterol. Vesicle hydration was calculated using two different methods and the agreement between them was excellent up to cholesterol concentration of 0.32 mole fraction. It was observed that the structural parameters change slowly with increasing cholesterol content up to around 0.3 mole fraction and a relatively abrupt structural alteration occurs above this cholesterol content. This abrupt structural change is consistent with the asymmetrical distribution of lipid composition between the inner and outer bilayer face.


Ionic strength induced structure in histone H4 and its fragments. The salt-induced folding and self-association of histone H4 and its fragments (1-23), (25-67), (69-84), and (69-102) have been studied at the same molar concentration (1 mM) by nuclear magnetic resonance (NMR), circular dichroism (CD), and ir spectroscopy. Byeach of these techniques intact histone H4 exhibited a fast structural change, involving the formation of alphaR helix and aggregation, and also a slow change involving beta-structure formation. Fragment (25-67) was found to behave in a manner similar to the intact molecule for the fast change, showing both helix formation and aggregation but exhibited no time-dependent effects. All the other fragments were found to be essentially noninteracting. It is concluded that (25-67) contains the rgion critical for the folding and self-association of histone H4. On the basis of these results a model is proposed for the self-association of histone H4 in which helix is located between residues 49 and 73, while the beta structure lies between 74 and the C-terminus.


The cytotoxins of cobra venoms. Isolation and partial characterization. Eight basic proteins which lyse virus-transformed mouse fibroblasts in culture have been isolated from the venoms of six Asian Naja naja subspecies. These cytotoxins appear to represent an homologous series of proteins, all within the molecular weight range of 7000-8000. They have been divided into three arbitrary types on the basis of amino acid composition, electrophoretic mobilities and elution order upon ion-exchange chromatography. The rate at which the toxins effect cell lysis: (1) appears to be a function of the basicity of each toxin; (2) is dependent upon toxin concentration; (3) is temperature dependent; and (4) is inhibited by heparin sulfate. In view of the physical changes, which the cell undergoes during lysis and of the various factors which affect the action of these proteins, it is proposed that interaction of membrane receptors with the toxin, leading to alteration of cell membrane structure, is the principal event which ultimately leads to the disruption of the cell.


Dynamic state of concanavalin A receptor interactions on fibroblast surfaces. Cultured normal and transformed fibroblasts were treated "in situ" by the concanavalin A-peroxidase labelling technique. It is known that peroxidase recognizes only a fraction of the bound lectin depending on the cell type. Kinetics studies revealed that 80 to 95 percent of the peroxidase and only 10 percent of the lectin are released from the cell surface when the labelled cells were reincubated at 37 degrees C. It is shown that it is mostly the concanavalin traced by peroxidase that is released and also that the lectin and the enzyme are shed as a complex or concomitantly. Consequently, the shedding pattern of the enzyme is used to demonstrate heterogeneity in the lectin binding sites; there are two main components labelled by concanavalin and peroxidase, one which has a short period (from 6 to 16 min) and another one with a much longer one (1.3 to 3 h). It is shown that when cells are incubated at 37 degrees C after a lectin treatment, secondary binding forces occur between the lectin and cell surface components which render the lectin unavailable for inhibiting sugars. Under the same conditions, some peroxidase can still be bound and a slight agglutination can still occur.