Righting the wrongs of writing: copy editors speak out.

The major part of APGA Press production editor's jobs is copy editing manuscripts before they are set in type. In this article, written at the invitation of the P&G editor, two APGA Press Staff members use specific examples to illustrate the grammatical and stylistic errors that cause problems for them and, if not corrected, for readers. The authors hope that contributors and potential contributors to professional journals will find this article helpful in expressing their ideas clearly when preparing material for publication.


Anesthetic considerations in diffuse bleeding diathesis of uncertain origin.

This is a report of a case of a diffuse bleeding tendency in a pregnant woman who presented for emergency splenectomy with a tentative diagnosis of thrombotic thrombocytopenic purpura. The influence of multiple organ dysfunction in the selection of appropriate monitors and the anesthetic technic in such cases are complex.


Bacteriologic aspects of epidural analgesia.

The incidence of contamination of catheters and syringes used during epidural analgesia for parturients and the effectiveness of bacterial filters were investigated. The effect of bupivacaine on bacterial viability and growth was also studied. Syringes in 5/101 cases were contaminated, while catheter tips located in the epidural space were sterile. Organisms isolated were skin commensals and probably originated on the hands of anesthetic personnel. Bupivacaine (0.25%) was bacteriocidal to S epidermidis and Corynebacterium spp at 37C but not at room temperature. These findings illustrate the efficacy of using bacterial filters during continuous epidural analgesia. New syringes should be used for each epidural injection as insurance against seeding of bacteria in the presence of a defective filter.


Cardiovascular dynamics after large doses of fentanyl and fentanyl plus N2O in the dog.

The effects of large doses of fentanyl (0.05 to 2 mg/kg) and fentanyl plus N2O on cardiovascular dynamics were determined in 10 unpremedicated dogs breathing 100% O2. Using computer analysis of the central aortic pulsepressure curve, stroke volume (SV), cardiac output, heart rate (HR), peripheral vascular resistance (PVR), and systolic, diastolic, and mean arterial blood pressures (BP) were determined while fentanyl was being given at a rate of 0.3 to 0.44 mg/min. Fentanyl caused a dose-related decrease in HR, which was significant at 0.05 mg/kg. Cardiac output, PVR, and systolic, diastolic, and mean arterial BP were also decreased and SV increased. The latter changes became significant at 0.1 mg/kg for diastolic BP; 0.15 mg/kg for cardiac output and mean BP; 0.25 mg/kg for sv and systolic BP; and at 1.25 mg/kg for peripheral vascular resistance. Addition of N2O after fentanyl did not significantly change any parameter, although SV, cardiac output, and HR were usually increased and PVR decreased. These data demonstrate that, while large doses of fentanyl or fentanyl plus N2O do alter cardiovascular dynamics in dogs, the changes appear to be less profound than those produced by equianalgesic doses of morphine. Our findings suggest that large doses of fentanyl-O2 may be an attractive alternative to morphine-O2 anethesia in critically ill patients.


Management of a parturient with cardiac valve prosthesis.

The parturient with a cardiac valve prosthesis presents a twofold problem of management: (1) that of a rising cardiac workload in the presence of serious heart disease, and (2) that of chronic anticoagulant therapy. Pain relief, a shortened 2nd stage of labor, and avoidance of bearing-down efforts reduce stress. Vaginal delivery is preferred to cesarean section, and segmental lumbar extradural block is the optimal anesthetic method. Oxytocics must be used with caution and heparin is preferred to longacting oral anticoagulants.


Survival following massive pulmonary hemorrhage complicating pulmonary embolectomy: a case report.

Continued employment of extracorporeal circulation provides for adequate oxygenation despite massive pulmonary hemorrhage. However, this modality requires continuing heparinization, which seems to preclude control of the hemorrhage. The longer bleeding persists, the more the pulmonary damage. Further, the problems of massive volume replacement will be added to this catastrophe. The authors believe that rapid discontinuance of extracorporeal bypass, reversal of heparinization with protamine, aggressive pulmonary suctioning, and alternate ventilation with 100% O2 were responsible for their good result. Arterial hypoxemia was avoided despite pulmonary suction yielding 1500 ml in 17 minutes.


Factors influencing choice between tracheostomy and prolonged translaryngeal intubation in acute respiratory failure: a prospective study.

One of the problems of prolonged ventilatory therapy in acute respiratory failure (ARF) is the need to choose between tracheostomy after 48 to 72 hours of translaryngeal (TL) tracheal intubation or the continuous use of the TL tube for a period of 10 days. Too often the choice has been based on retrospective studies or personal preference. To investigate this problem prospectively, 52 adults in ARF were divided sequentially into 2 groups on their 3rd day of TL intubation. Patients in group I (G-I) retained the TL tube for a total of 11 days; those in group II (G-II) were tracheostomized on the 3rd day. The following factors ere used to evaluate the efficiency and complications in each group: patient's epidemiologic variables, daily pulmonary functions, severity of respiratory infections, and scores of post-intubation airway lesions. No consistent statistically significant differences between the two procedures were seen in the pulmonary functions or the range of individual patient variables. However, with an early tracheostomy, there was an eightfold greater incidence of contamination of the airway by new organisms, airway lesions were more frequent and severe, and the need for the tracheal tube was extended. To identify the epidemiologic variables and the pulmonary functions that discriminate between patients with serious airway lesions and those with mild lesions, and to evaluate the ability of these variables to differentiate the patients who died from those who survived, the distribution of all factors was compared in the two categories. The epidemiologic variables separated the patients according to their airway lesions only, while the difference in pulmonary functions was statistically significant only between the patients who died and those who survived.


Atypical pseudocholinesterase: a clinical report.

The safe conduct of anesthesia demands careful evaluation of the previous experience of patients under anesthesia and surgical operation, as well as that of their families. Untoward responses that may recur with subsequent anesthetic procedures should be documented and patients informed, as the need dictates. Two cases of prolonged apnea following single intubating doses of succinylcholine could not have been predicted by the patient's preanesthetic history. As a result of these problems, the authors recommend more careful preanesthetic history-taking with reference to patients' prior anesthetic experiences.


Management of intractable pain in adiposis dolorosa with intravenous administration of lidocaine.

Intractable pain in a patient with adiposis dolorosa (Dercum's disease) was treated by IV administration of lidocaine (200 to 400 mg). Relief was maximum 20 minutes after the end of drug infusion and persisted for over 10 hours. Toxicity was minimal. Slow EEG waves which appeared during drug administration disappeared within 20 minutes.


Resection of stenotic trachea: a case presentation.

Tracheal stenosis is usually a complication of prolonged mechanical ventilation. If the narrowing is severe it will greatly obstruct the air flow. Resection of the damaged trachea is a life saving procedure. The anesthetic management of the resection of an "ice cream cone narrowing" of the trachea (3 mm diameter) 4 cm above the carina is reported. A forced vital capacity loop (expiration-inspiration) of air flow versus volume revealed preoperatively little difference in the flow during the forced (F-V) and resting ventilation (Vt), thus confirming a diagnosis of severe upper airway obstruction. A total obstruction of the air flow was encountered during the skeletonization of the trachea and the surgeon was allowed to work for not more than 60 seconds at a time. Perfect cooperation between the surgeon and the anesthetsia team is necessary for the successful completion of this type of surgery.


Failure to induce hepatic pathology in animals sensitized to a halothane metabolite and subsequently challenged with halothane.

The results of this animal study confirm the induction of delayed hypersensitivity to a halothane metabolite, but the sensitization does not cause hepatic damage on subsequent halothane exposures. Before the hypothesis that hepatic necrosis following halothane has an immunologic basis can be accepted or rejected, more critical studies with different approaches must be made.


Anesthetic considerations in patients with gas gangrene.

Anesthesia for a patient with gas gangrene presents a challenge for the anesthesiologist, since it is an uncommon disease requiring emergency treatment. The authors, faced with such a challenge and finding little guidance in the literature, have proposed modalities of anesthetic management based on pathophysiology, symptomatology, and the reported experience of others. In addition to choice of anesthetic agents, problems reviewed include shock, hypovolemia, tachycardia, fever, anemia, renal dysfunction, pulmonary insufficiency, and contamination. Factors relating to anesthesia during hyperbaric-O2 therapy are also reviewed.


Circulatory changes in patients with coronary artery disease following thiamylal-succinylcholine and tracheal intubation.

Circulatory responses after thiamylal (4 mg/kg) and succinylcholine (SCh) (2 mg/kg) administration followed by direct laryngoscopy and tracheal intubation were measured in 20 patients before elective aortocoronary vein bypass graft operations. Compared with awake measurements, the mean arterial pressure (MAP) decreased 19 +/- 3 torr (mean +/- SE) and heart rate (HR) increased 9 +/- 3 bpm 1 minute after thiamylal-SCh. MAP was increased 39 +/- 4 torr and HR 20 +/- 3 bpm above awake levels in response to laryngoscopy and tracheal intubation. Blood pressure and HR returned spontaneously to near awake levels without additional anesthesia within 5 minutes of anesthetic induction. Stroke volume index was decreased significantly after tracheal intubation but cardiac index was not altered. The authors conclude that thiamylal-SCh followed by tracheal intubation is an acceptable anesthetic induction sequence for patients without evidence of left ventricular heart failure who require anesthesia for elective coronary artery revascularization.


A comparative study of enflurane and halothane using systolic time intervals.

The effects of enflurane and halothane anesthesia on systolic time intervals were studied in 12 healthy patients. Cardiovascular measurements were made at equipotent levels of anesthesia: enflurane 1.23% end-tidal and halothane 0.65% end-tidal. These agents were studied first with 50% N2O and then without N2O. Systolic, diastolic, and mean arterial blood pressure were decreased more by enflurane than by halothane (p less than 0.05). However, halothane caused significantly more myocardial depression than enflurane, as indicated by a larger preejection period (PEP) and preejection period/left ventricular ejection time (PEP/LVET) and a smaller 1/PEP2 and ejection fraction. When N2O was discontinued, both agents increased PEP and PEP/LVET and decreased 1/PEP2 and the ejection fraction.


Effect of PaCO2 on O2 consumption during cardiopulmonary bypass in man.

Under conditions of anesthesia with controlled respiration, the changes in the state of the circulation due to altered PaCO2 are often overlooked or attributed to the anesthetic agent. During cardiopulmonary bypass when cardiac output is kept constant, change in fluid level of the extracorporeal reservoir reflects overall change in the circulatory beds. Following an hour on total bypass for stabilization of all parameters, a change in the oxygenator ventilating-gas mixture was induced to increase or decrease the PaCO2 an average of 11 torr in 40 patients. A reduction in PaCO2 from 37 to 25 torr increased mean arterial blood pressure (MAP), central venous pressure (CVP), and the volume of blood stored in the extracorporeal circuit, signifying an overall reduction in the intravascular capacity. This was associated with a decrease in total O2 consumption (VO2). An abrupt increase of PaCO2 from 26 to 37 torr produced a decrease in MAP,in CVP, and in the reservoir blood level, signifying blood "take up" by the patient. There was also an associated increase in VO2.


The effects of doxapram on cerebral blood flow and peripheral hemodynamics in the anesthetized and unanesthetized goat.

The effects of doxapram on cerebral blood flow (CBF) and peripheral hemodynamics were evaluated in the goat after direct injection into the cerebral circulation and after peripheral IV administration. Doxapram injected centrally via the temporal artery, in doses too small to affect peripheral circulation, produced an immediate and prolonged decrease in CBF in both anesthetized and unanesthetized gohanges in blood CO2 concentration, since blood flow decreased immediately after injection and as reduction in CBF occurred in the absence of changes in blood CO2. Pheripheral IV administration of doxapram in clinical doses to anesthetized and unanesthetized goats produced a biphasic cardiovascular response. Cerebral blood flow, cardiac output, blood pressure, and heart rate decreased abruptly, but with the exception of CBF, these parameters returned to or exceeded preinjection values within 30 seconds. Cerebral blood flow slowly returned to preinjection values (within 10 minutes) and, in the case of unanesthetized animals and anesthetized animals ventilated with room air, rose substantially above preinjection levels, reaching a peak 20 minutes after injection. The latter observation is thought to be a compensatory reaction to the reduction in CBF; it was not evident in anesthetized animals ventilated with 100% O2.


Hyposensitization therapy with alum-precipitated pyridine extracts in animal dander sensitive patients.

Hyposensitization therapy with APP extracts was found to be effective in 80% of 185 patients allergic to animals (96 to cats, 84 to dogs and 5 to horses) with a minimum of reactions (2.5 local and 0.2% constitutional). Lower respiratory symptoms were the most common complaints, followed by nasal, ocular and respiratory, singly or in combination.


Development of Ehrlichia canis, causative agent of canine ehrlichiosis, in the tick Rhipicephalus sanguineus and its differentiation from a symbiotic Rickettsia.

Certain aspects of the development of Ehrlichia canis, causative agent of canine ehrlichiosis (tropical canine pancytopenia) in Rhipicephalus sanguineus ticks were studied. It was found that partial feeding of nymphs infected as larvae with E canis was a desirable, if not necessary, preliminary treatment for successful infection of dogs with ground-up ticks. It remains unclear whether feeding increased the number or altered the virulence of ehrlichiae within tick tissues. Ehrlichia canis organisms were detected by immunofluorescent microscopy in the midgut and hemocytes and by electron microscopy in the midgut and salivary glands of partially engorged adult ticks which had been infected as larvae and nymphs. Organisms were not observed in the ovary. Intracytoplasmic inclusions contained 1 to 80 elementary bodies, each provided with 2 distinct membranes. Infection of the midgut and salivary gland was confirmed by injecting homogenates of these tissues into susceptible dogs. Staining of gut smears of partially engorged adult ticks by fluorescein-conjugated anti-E canis antibody was found to be a reliable indicator of the infection.


Intestinal immune response of feeder pigs to infection with transmissible gastroenteritis virus.

Five feeder pigs 4 to 6 months old were orally inoculated with transmissible gastroenteritis virus. Diagnosis of transmissible gastroenteritis was made on the basis of clinical signs and examination of intestinal mucosa by the fluorescent antibody technique. Immunoglobulins were extracted from intestinal fluid of infected feeder pigs. Virus-binding and neutralizing antibodies were detected in intestinal extracts between 7 and 56 days after infection. The concentration of binding antibodies reached a peak at 21 days after infection and was on the decline at the end of the experiment on the 56th postinfection day. In contrast, neutralizing intestinal antibody concentration was increasing on day 56. In both systems, the predominant immunoglobulin was of the IgA class. Examination of blood serums of the pigs by the plaque-reduction technique showed progressive antibody increases ranging in titer from 1:8 on day 7 to 1:256 on day 56 after infection. An analysis of the protein profiles from these serums showed a significant increase in the concentration of gamma-globulins and a decrease in the albumin fraction.


Characterization of miniature pig kidney cells and their resistance to chlamydial infection.

A miniature pig kidney cell line has been established from porcine fetuses taken aseptically by hysterectomy and maintained for more than 50 passages in Eagle's minimal essential medium containing 10% heat-inactivated newborn calf serum. Cell transfers were performed each week. Primary and serially passaged cells were found to be highly refractory to infection by Chlamydia trachomatis strains TW-3, Bour, and LGV 440L and Chlamydia psittaci strains meningopneumonitis and 6BC and insusceptible to poliovirus type 1. The cells were susceptible to vesicular stomatitis virus and herpes simplex type 1 virus. Identity of the cells was established by cytotoxicity, isozyme, and cytogenetic studies.


Etomidate: an ultrashort-acting nonbarbiturate agent for anesthesia induction.

Clinical evaluation of etomidate, an ultrashortacting nonbarbiturate hypnotic agent, shows it to be useful for anesthetic induction in adults. It produces sleep in 1 arm-brain circulation time, metabolites apparently peaking at 7 minutes postinjection. Cardiovascular and respiratory systems appear minimally affected and there is no indication of organ toxicity or other biochemical or hematologic drug-induced disturbances. Moderate pain on injection at some sites and transient myoclonia were the principal disadvantages observed.


Estrogen- and progesterone-induced uterine motility of anestrous goats.

Uterine motility was studied in the anestrous goats after a series of estradiol injections was given and after estradiol-plus-progesterone treatment was administered. Estradiol (0.1 mg/kg) given alone by intramuscular route every 6 hours for 4 continuous days produced only a moderate increase in the uterine contractions during treatment, but significantly higher amplitude and frequency of contractions occurred at 2 days after treatment was stopped. The same doses of estradiol administered for a 24-hour period and followed by 4 injections of progesterone (1.0 mg/kg) given intramuscularly once a day produced a moderate response during treatment, but depressed the amplitude and the frequency of contractions after the treatment was stopped. The results indicate that the tissues primed with estrogen produced a stimulatory effect after their estrogen contents diminished. Progesterone, likewise, produced inhibition after a decrease in the progesterone contents of the uterine tissue.


Erysipelas arthritis in swine: lysosomal enzyme levels in synovial fluids.

Arthritic and histologically normal joints from swine in which arthritis had been produced by the intravenous inoculation of Erysipelothrix rhusiopathiae were used as a source of fluids for lysosomal enzyme determinations. Mean lysozyme activities in synovias from arthritic and histologically normal joints were 16.60 and 5.79 mug/ml, respectively. Acid phosphatase (ACP) was increased more than 8 times the activity in histologically normal joints, but there was no relationship between lysozyme and ACP, indicating the probability that 1 of these enzymes came from another source. The cytoplasmic enzyme, lactic dehydrogenase (LDH), was increased in proportion to ACP, indicating that cell death and not selective extrusion of lysosomal enzymes during phagocytosis was an important mechanism of enzyme release in arthritic joints. Lysozyme activities in synovias from histologically normal joints were often increased above companion serum concentrations, indicating the enzyme has a special role in the joint. Also, the ratio of activities of lysozyme to ACP in pig buffy coat lysates was different from the ratio of the 2 enzymes in synovias from arthritic joints.


The influence of suction catheter tip design on tracheobronchial trauma and fluid aspiration efficiency.

The suctioning efficiency and trauma-producing characteristics of five commercially available tracheobronchial suction catheters (Pharmaseal Tri-Flo, NCC Gentle-Flo, Argyle Aero-Flo, Argyle Dual Side-Hole, and Pharmaseal Whistle-Tip) were experimentally evaluated in anesthetized healthy dogs. The tendency of catheters to invaginate or "grab" tracheobronchial mucosa was observed with a bronchofiberscope during suctioning. Mucosal grabbing was seldom seen even at high (greater than 300 torr) vacuum levels with the cateter tip in the trachea. All catheters were observed to invaginate mucosa in lobar and segmental bronchi, with the frequency of grabbing being a function of airway anatomy, airway size, catheter orientation, tip design, and vacuum level. Catheters with multiple side-holes appeared to invaginate mucosa less frequently than the single side-hole catheter. Repeated suctioning of anesthetized healthy dogs followed by tracheobronchial excision, gross observation, and histologic examination of mucosal tissue biopsies demonstrated significant differences in the frequency and severity of lesions caused by the tracheobronchial suction procedure. All catheters were observed to damage airway lining, the damage related to multiple side-hole catheters appearing to be associated entirely with the act of cateter insertion and not with the application of vacuum. Only the Whistle-Tip design produced measurable damage beyond that related to catheter insertion. The average tip-suctioning effectiveness for each catheter, determined in vitro by aspirating a thin, uniform layer of simulated mucus, was found to be significantly higher for the Tri-Flo and Whistle-Tip catheters than the others, the Aero-Flo being least effective. Preliminary attempts to demonstrate this difference in suctioning effectiveness by comparing the performance of the catheters which displayed the highest and lowest tip suction effectiveness in a standardized clinical suctioning procedure revealed no significant difference in the percentage of mucus removed by either catheter. Additional studies should clarify this apparent contradiction.


Sphincteroplasty in selected cases of acute recurrent pancreatitis.

Although most patients with acute pancreatitis can be treated sucessfully by non-operative therapy and progress to oral alimentation without difficulty, an occasional patient may require exploration during the acute or subacute phase. This is justified principally by the recurrence of pain following attempts at oral alimentation in the hospital.


The continent ileostomy and the isolated ileal bladder.

The histories of 19 patients operated on by Kock's technique are reported. Follow-up extended from 3 years 4 months for the first case to 2 weeks for the last. There was one fatality. Some details of technique, contraindications, and complications and their treatment are discussed. Seventeen patients had a total proctocolectomy for ulcerative colitis and 2 had urinary diversion into an isolated ileal reservoir for inoperable carcinoma of the urinary bladder.


Quantitative measurement of smooth pursuit eye movements.

Smooth pursuit eye movements were quantitatively assessed in 25 normal subjects and 22 patients. A laboratory digital computer was used to compute 200 eye velocity samples per second and to statistically compare these eye velocity measurements for five different object velocities. Of six statistics evaluated, mode eye velocity showed the least variability in normal subjects and was most frequently abnormal in patients. Compared to normal subjects, patients with brain stem degeneration and cerebellar-pontine angle tumors with brain stem compression had significant impairment of smooth pursuit. Patients with peripheral vestibular lesions and C-P angle tumors without brain stem compression did not have impaired smooth pursuit. These preliminary findings suggest that quantitative measurement of pursuit eye velocity can be a sensitive test for brain stem dysfunction.


Secretory lysozyme of the human middle ear mucosa: immunocytochemical localization.

Lysozyme was demonstrated by an immunocytochemical technique in the biopsied mucosa obtained from the promontory of the fifteen patients who had chronic middle ear effusions. Lysozyme was localized in the mucigen granules of the secretory cells, as well as in the specific granules of the polymorphonuclear neutrophilic leukobytes (PMN) and macrophages. The specimens obtained from patients with mucous effusion showed numerous secretory cells that contained lysozyme, in sharp contrast to the serous type in which only a few secretory cells could be found. The present morphological finding was in agreement with the biochemical finding which demonstrated higher lysozyme level in mucous effusions than that of the serous type. It was concluded that human middle ear mucosa provided lysozyme and that its secretion was active in serous otitis media, particularly of mucoid type.


Hereditary abductor vocal cord paralysis.

Familial bilateral abductor cord paralysis was described in the father and two sons of a family in which the ramaining siblings (obe boy and three girls) were normal. The onset of stridor ranged from six months to nine years after birth in these patients who were all treated with a tracheostomy. Normal development and intelligence was experienced by all three patients. Since previous reports  of hereditary abductor cord paralysis described mental retardation in all patients who were maintained without tracheostomy, anoxia from unrelieved laryngeal obstruction may be a significant complication following nonsurgical management of patients who appear to tolerate bilateral abductor vocal cord paralysis.


Hearing function and chronic renal failure.

In order to evaluate the degree and type of hearing loss in patients with chronic renal failure, 61 patients undergoing chronic hemodialysis were examined. Hearing threshold levels, adjusted for age and sex, demonstrated a significant high frequency deficit, which in some patients was noted early in the course of hemodialysis. Fluctuations in hearing were noted over single dialysis events but were transient and apparently independent of corresponding changes in Na, K, Ca, BUN, creatinine, glucose, mean blood pressure, and weight. Preliminary attempts to evaluate hyperlipidemia as a possible cause of hearing loss did not reveal any hearing deficit or gain as related to triglyceride or cholesterol levels. The data suggested other possible causes of observed auditory loss.


Nasal-cardiopulmonary reflexes: a role of the larynx.

A brief review of the literature from otorhinolaryngology and the basic sciences shows the existence and role of nasal-cardiopulmonary reflexes in animals and man. There is ample evidence that odors, fluids and mechanical stiumlation of the nasal mucosa will induce changes in the lungs and cardiovascular system. The proposition that nasal obstruction also produces cardiopulmonary changes is briefly reviewed. The suggestion is made that one of the functions of the nose is to act as an expiratory brake. the removal of this brake could result in changes in laryngeal resistance that lead to poor ventilation.


Chorionic somatomammotrophin as index of fetal growth.

Concentrations of chorionic somatomammotrophin (hCS) in maternal plasma at the time of delivery and in placental extracts were determined by radioimmunoassay and related to neonatal growth in 111 normal pregnancies. Mean maternal plasma hCS concentration was lower in association with small-for-dates infants, defined by either height or weight, than with well-grown preterm or term infants. Mean placental hCS concentration was not decreased in association with undergrown infants though placental hCS was correlated with maternal hCS (r=0 - 245).


Phototherapy for hyperbilirubinaemia in very low birthweight infants.

Phototherapy was used to treat 20 newborn babies whose birthweight was below 1500 g and whose plasma bilirubin exceeded 8 mg/100 ml. The plasma bilirubin level was maintained below 13 mg/100 ml except in 4 babies whose level exceeded 13 mg/100 ml before treatment was started. In 60% of an untreated group of larger babies previously reported the plasma bilirubin level exceeded this figure. Phototherapy seems to control the plasma bilirubin level satisfactorily in very low birthweight infants, but frequent measurements on the second and third days of life are advised in order that treatment may be started promptly when it exceeds 8 mg/100 ml.


Clinical comparison between glucose and sucrose additions to a basic electrolyte mixture in the outpatient management of acute gastroenteritis in children.

In a double-blind trial in 94 children attending outpatients the value of glucose or a sucrose addition to a basic electrolyte mixture for the management of acute gastroenteritis was compared. Of the children treated with added sucrose 10% failed to respond compared with 27% of those treated with added glucose. This difference was significant (P=0-05), but the time to recovery in those in the two groups who responded to treatment was not significantly different. Thus, despite theoretical advantages, there was no practical advantage in using glucose rather than sucrose. A 5% sucrose electrolyte solution with its relatively low osmolality, ready availability, and ease of preparation is recommended as the treatment of choice in the outpatient management of acute gastroenteritis in infancy.


Recurrent thrombocytopenic purpura associated with accessory spleen.

In the case of a boy with a relapsed thrombocytopenic purpura removal of a splenunculus, shown by radioisotope scanning, resulted in a partial remission. The significance of the association of relapse in ITP and the presence of splenunculi remains unknown: investigation of similar cases would clarify the matter.


Clinical studies in alcoholic sideroblastosis.

The incidence and characteristics of ring sideroblastic and megaloblastic changes in bone marrow were studied in chronically ill, malnourished alcoholics and well-nourished alcoholics without complicating medical illness. A clear correlation of blood alcohol level with changes in serum pyridoxal-5-phosphate (PLP) values or with the incidence of ring sideroblasts could not be demonstrated. The appearance of ring sideroblasts was associated with dietary restrictions of pyridoxine and noticeable folic-acid-deficient megaloblastosis. The majority of subjects with ring sideroblasts were from the chronic, malnourished alcoholic group; the number of sideroblasts correlated with severity of marrow magaloblastic change. While low serum PLP levels were characteristic of the chronically ill alcoholic with ring sideroblasts, equally low levels were detected in the absence of the marrow abnormality. Thus, a low serum PLP value alone is not a certain indication of the presence of marrow ring sideroblasts.


[Thielavia heterothallica spec. nov. (Abb.1) Status conidialis: Chrysosporium thermophilum (Apinis) von Klopotek].

Thielavia heterothallica spec. nov. is described as a heterothallic, thermophilic fungus with spherical, black, non-ostiolate cleistothecia; elliposidal evanescent asci which contain eight one-celled ellipsoidal ascospores, darkening to deep brown to black, with one germ pore. The conidial state is Chrysosporium thermophilum (Apinis) von Klopotek.


A behavioral paradigm for the evaluation of narcotic antagonists.

We have developed an experimental paradigm for the behavioral evaluation of narcotic antagonists. The study specifically examined the heroin-seeking behavior of hard-core narcotic addicts on a research ward under blocked and unblocked conditions. Each patient served as his own control. A long-term follow-up program in the community, with aftercare services, was utilized to determine the relationship between behavior observed on the research ward and behavior that occurred in the community. While preliminary one-month follow-up data offered some cause for an optimistic view of narcotic antagonist treatment, behavioral data observed on the research ward raised serious doubts about the possibility of extinguishing heroin self-administration with antagonists. The behavioral data were not consistent with laboratory descriptions of extinction. Rather, the data suggested that narcotic antagonist programs should emphasize the development of contingencies for the reinforcement of narcotic antagonist self-administration to ensure an opiate-free state, instead of focusing on an extinction approach.


Pharmacologic studies on anti-inflammatory activity of di- and triketopiperidine derivatives.

A number of derivatives of piperidine-2,4,6-trione and oxazine-2,4-dione with cyclohexyl and allyl substituents at their ring were evaluated pharmacologically. Piperidine-2,4,6-trione compounds, regardless of type of substituent, were readily absorbed from the site of their introduction and displayed similar biological activity. Introduction of the N-cyclohexylcarboxamide substituent diminished general toxicity of the derivative and increased anti-inflammatory activity in all tests. The oxazine-4,6-dione derivatives were poorly absorbed. The bis-(1-cyclohexyl-5,5-diallyl-piperidine-2,4,6-trione) derivative, product of condensation of two molecules of a simpler compound, was very well absorbed, strongly toxic, and showed distinct anti-inflammatory activity. Limited correlation between chemical structure and biological activity was noted, supporting the concept that introduction of cyclohexyl and allyl radicals imparts anti-inflammatory and immunosuppressive activity to derivatives of this type.


System-structured management of acutely ill surgical patients.

The management of surgical patients requiring intensive care is complicated by abnormalities in multiple body systems. An effective method of organizing data collection and treatment is essential. The widely applied "problem-oriented" approach is not entirely satisfactory in the context of surgical intensive care. An alternative method is presented in which management is structured by body systems. The function of each system is analyzed and a plan for each is defined. Data display is arranged by systems rather than problems and is compatible with modern computer technology. A surgical patient under treatment for pancreatitis is presented to illustrate this approach. Experience with 220 patients on a residency teaching service supports the impression that system-structured management is an effective method of managing the course of an acutely ill patient and should be widely applicable to such patients in any general surgical environment.


Primary diffuse microscopical hyperplasia of the parathyroid glands: surgical importance.

In two of 182 patients with verified primary hyperparathyroidism, microscopical hyperplasia was present in all parathyroid glands that were normal in size or only slightly enlarged. All parathyroid glands in another two patients showed microscopical hyperplasia and varied from a normal size of 190 mg. In seven additional patients, microscopical hyperplasia was present in one, several, or all parathyroid glands, which varied in weight from normal to 350 mg. Familial hyperparathyroidism or multiple endocrine neoplasia was evident in five of 11 patients. Contributing to difficulties was the experience in five patients in whom removal of mildly enlarged parathyroid glands corrected hypercalcemia, but definite microscopical abnormalities were not evident by routine histologic study of the glands. Thus, there appears to be a spectrum of abnormalities relative to size and microscopical changes in parathyroid glands of patients with primary hyperparathyroidism. The surgeon should be aware of these patterns of parathyroid hyperplasia that require a search for a fifth parathyroid gland and a subtotal parathyroidectomy.


Accidental persistent infection of cell lines by Newcastle disease virus, showing three unusual features--defective neuraminidase, temperature sensitivity and intranuclear inclusions.

A persistent, defective infection by an unknown strain of Newcastle disease virus (NDV) appeared accidentally in established lines of pig, ox and sheep kidney cells. Virus particles released from the persistently infected cells were not infectious and were deficient in neuraminidase activity. Synthesis of some of the virus-specified proteins in the persistently infected cells was temperature-sensitive. Co-cultivation of mixed populations of carrier cells and healthy chick embryo cells induced cell fusion with the formation of multinucleate heterokaryons and intra-nuclear inclusions. The development of inclusions in the chicken nuclei was not accompanied by 'rescue' of infectious NDV.


Histochemical studies in experimental portal-systemic encephalopathy.

Results of a histochemical study of glutamic dehydrogenase in experimental portal-systemic encephalopathy with hyperammonemia indicate that the enzyme's activity in brain was increased in all stages of the encephalopathy, and this increase appeared to be localized exclusively in astrocytes. The results are consistent with the view that the astrocyte has a critical role in ammonia metabolism in brain, probably in ammonia detoxification. The findings, moreover, indicate that one pool of glutamate, possibly the small pool, is located in the astrocyte.


Hearing under stress: III. The effect of external auditory meatal pressure on speech discrimination.

It has been well established that unequal air pressure across the tympanic membrane causes an increase in puretone thresholds. Very little information is available, however, concerning concommitant effects on reception and discrimination of speech material. This study was designed to determine whether or not further detrimental effects upon the communication process might occur in the form of decreased speech discrimination ability. The findings established that a high positive pressure in the external meatus can result in a deterioration of the individual's ability to discriminate speech sounds.


Assignment of a gene for tryptophanyl-transfer ribonucleic acid synthetase (E.C. 6.1.1.2) to human chromosome 14.

We describe a simple method for locating tryptophanyl-tRNA synthetase (E.C. 6.1.1.2) on cellulose acetate gels (Cellogel) following electrophoresis. Employing electrophoretic conditions which result in the separation of mouse and human tryptophanyl-tRNA synthetase, we have analyzed extracts of a number of independently derived mouse-human somatic cell hybrids and subclones derived from these hybrids for the presence of human tryptophanyl-tRNA synthetase. Electrophoretic patterns of hybrid extracts which contain human tryptophanyl-tRNA synthetase exhibit three bands. This is consistent with published evidence that the enzyme for mammalian cells is a homologous dimer. The electrophoretic patterns derived from some hybrids are unusual in that the human and hybrid bands of activity are more intense than the mouse band from the same hybrid. An analysis of hybrid cells and extracts indicates that human tryptophanyl-tRNA synthetase segregates with human chromosome 14 and with the only enzyme marker which has previously been assigned to this chromosome, nucleoside phosphorylase.


Cardiomyopathy complicated by left ventricular aneurysms in children.

Ventricular aneurysms in children are unusual. Three patients with cardiomyopathy associated with angiographically proved left ventricular aneurysms in this age group are reported. Two of them were girls. The ages were 20 months, 7 years, and 14 years. Heart failure was present in all patients. There was radiological evidence of cardiomegaly in all three, and the electrocardiogram showed signs of necrosis in two of them. Selective left ventricular angiography disclosed generalized hypokinesis in all patients. One child had an aneurysm of the diaphragmatic wall. In another the aneurysm was localized in the muscular ventricular septum, causing severe subpulmonary stenosis by encroaching in the right ventricular outflow tract during systole. The third patient had an aneurysm of the left ventricular free wall partly encircling the left ventricle. The coronary arteries appeared normal in all cases. The clinical features of the underlying disease were not altered by the presence of the aneurysm except in the patient with the septal aneurysm and subpulmonary stenosis. In this patient the aneurysm was successfully resected.


Angiosarcoma of pericardium. Problems in diagnosis and management.

The clinical histories of two patients with angiosarcoma of the pericardium are described. Both were previously well young men who presented with cardiac tamponade caused by haemorrhagic pericardial effusions. If a pericardiectomy is undertaken for diagnosis, wide excision of the pericardium is recommended. This will provide a large specimen for histological examination and may prevent subsequent development of constriction.


A retrospective study of renal cancer with special reference to coffee and animal protein consumption.

Interviews were obtained with 106 patients with adenocarcinoma of the renal parenchyma, 33 patients with carcinoma of the renal pelvis and 139 individually matched control patients. Comparison of the cancer patients with the control patients showed no evidence of a positive association between either type of renal cancer and coffee or animal protein consumption. Carcinoma of the renal pelvis was associated positively with cigareete consumption (relative risk estimate 1-8) and the daily consumption of analgesic tablets was more frequent in patients with cancer of the renal parenchyma than in their matched controls (14-2% compared with 1-9%,P less than 0.005). It appeared likely that the latter relationship was non-causal.


A pathogenetic model for erosive synovitis: lessons from animal arthritides.

Adjuvant arthritis, streptococcal cell wall-induced arthritis, and Erysipelothrix insidiosa arthritis are laboratory models of relapsing, erosive synovitis. A review of the experimental literature suggests that their pathogenesis is similar. The persistence in macrophages in the pannus of nonbiodegradable microbial cell wall components containing the peptidoglycan moeity is the central event. Based on this experimental literature, a model for the pathogenesis of these arthritides is developed. This model is testable and relevant to erosive synovitis in man.


The diurnal rhythm of enzymes in human red cells.

The activities of glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate: NADP oxidoreductase, G6PD), 6-phosphogluconate dehydrogenase (6-phospho-d-gluconate: NADP oxidoreductase, 6PGD), hexokinase (ATP:D-hexose 6-phosphotransferase, HK), lactic dehydrogeanse (L-lactate: NAD oxidoreductase, LDH) and aspirate aminotransferase (L-aspartate: 2-oxoglutarate aminotransferase, Asp.T) were determined in red blood cells of 11 healthy individuals. The determinations were carried out on samples drawn every 4 h over a 24 h period. The activities of G6PD, 6PGD, LDH and Asp.T exhibited a semi-circadian rhythm, namely, two peaks of activity during 24 h while HK activity demonstrated a true circadian rhythm. In addition a polymorphism of the G6PD and LDH activity patterns was observed. The implications of a biological clock in enucleated cells are discussed.


RNA polymerase activity and uterine growth: Differential stimulation by estradiol, estriol, and nafoxidine.

We have shown previously that estradiol, estriol, and Nafoxidine (Upjohn 11, 100A) have differential effects on uterine growth and that these effects are associated with the retention of the estrogen receptor by the nucleus. In order to examine these relationships further, we have studied the effect of these hormones on endogenous nuclear RNA polymerase I and II in the immature rat uterus. All three compounds caused a rapid elevation in polymerase II activity that reached a peak by the first hour and declined to almost control levels by 2 h after the injection. This transient peak in polymerase II activity was followed by a second elevation by the fourth hour in estradiol- and Nafoxidine-treated animals which was not observed in estriol-treated rats. The activity of polymerase I increased monotonically to very high levels by 4 h and was maintained 12 h or longer in estradiol- and Nafoxidine-treated animals. A similar elevation was observed in estriol-treated rats but the activity declined very rapidly to control levels by 12 h. The second elevation in polymerase II activity and the sustained stimulation of polymerase I activity were correlated with the stimulation of true uterine growth. These data confirm our previous suggestion that long-term nuclear retention of the receptor is a requirement for true uterine growth and suggest that an obligatory response in the production of true growth is the stimulation of a second rise in polymerase II activity and an elevated and sustained activity of polymerase I.


Photochemical cross-linking of neighboring residues in protein-nucleic acid complexes: rnase and pyrimidine nucleotide inhibitors.

Iradiation of the stable complexes formed between RNase and its competitive inhibitors cytidine 2'(3'),5'-diphosphate (pCp), and uridine 2'(3'),5'-diphosphate (pUp), resulted in covalent bond formation between the pyrimidine nucleotides and the enzyme. The photochemical reactions were initiated by ultraviolet light of lambda greater than 300 mn, employing acetone as a photosensitizer. This method was found to yield less undesired by-products, particularly photolyzed amino acids and aggregates resulting from protein-to-protein cross-linking, than the direct method of irradiation with light of lambda 260 nm. Tryptic digrestion of the modified protein, and chromatographic analysis of the peptides thus obtained, revealed a single and specific peptide which bacame covalently linked to both nucleotide inhibitors. The amino acid composition of this peptide is consistent with the sequence Asn-67-Arg-85 of RNase. Part of this peptide (residues 78 through 83) is close to the enzyme's binding site for the pyrimidine moiety of the nucleotides. Denatured RNase failed to cross-link to the inhibitors, and the extent of pUp cross-linking could be reduced by the addition of another competitive inhibitor (3'-UMP). Finally, the presence of excess inhibitor in the irradiation mixture did not lead to nonspecific cross-linking. This indicates that specificity is also achieved due to the fact that unbound excited inhibitor molecules do not react with the protein but prefer to follow different and faster reaction paths.


Cryoenzymology of chymotrypsin: the detection of intermediates in the catalysis of a specific anilide substrate.

The reaction between chymotrypsin and N-acetyl-L-phenylalanine p-nitroanilide has been studied at subzero temperatures in fluid aqueous dimethyl sulfoxide solvent. Following initiation of the reaction at temperatures as low as -90 degrees C, a series of four reactions prior to the normal rate-limiting step (acylation) was detected spectrophotometrically. Various experimental observations have led to the following interpretation of these reactions. Reaction 1 corresponds to the binding of substrate yielding the initial Michaelis complex. Reactions 2 and 3 are two pH-independent reactions, ascribed to substrate-induced changes in the positions of active-site groups. Reaction 4 is a pH-dependent reaction (pK = 5.9) which involves the imidazole of His-57 but which is not the formation of a tetrahedral intermediate, oxazolinone, or acyl enzyme. The slowest detected step corresponded to the acylation reaction. No evidence for the accumulation of a tetrahedral intermediate was obtained. Spectral, kinetic, and thermodynamic data for these reactions are presented, as is justification for the relevance of these findings to the reaction under physiological conditions. These results demonstrate the utility of subzero temperatures in enzyme mechanism studies, especially with regard to allowing the accumulation of intermediates which may be quite stable at appropriate values of pH and low temperature.


Levels of RNA polymerases during the mitotic cycle of Physarum polycephalum.

Two RNA polymerase activities were quantitatively solubilized in plasmodial homogenates from Physarum polycephalum by sonication at 0.5 M ammonium chloride concentration. The proportions of RNA polymerases A and B were determined by four different methods. Equal activity levels of both enzyme A and enzyme B were detected throughout the synchronous mitotic cycle of Physarum.


Inhibitors of protein synthesis in eukarytic cells. Comparative effects of some amaryllidaceae alkaloids.

The effects of eighteen compounds obtained from bulbs of the Amaryllidaceae family were tested on (a) animal cell growth, (b) DNA, RNA and protein synthesis by intact cells and (c) protein synthesis in cell-free systems. Dihydrolycorine, haemanthamine, lycorine, narciclasine, pretazettine and pseudolycorine halted HeLa cell growth at 10(-1) mM or lower concentrations. These compounds at their growth inhibitory concentrations block protein synthesis in ascites cells and stabilize HeLa cell polysomes in vivo. Endomyocarditis virus RNA-directed cell-free polypeptide synthesis by an ascites S-30 extract and acetyl-[14C]leucyl-puromycin formation by ascites ribosomes are also inhibited by the six compounds indicated above. It is therefore concluded that they halt protein synthesis in eukaryotic cells by inhibiting the peptide bone formation step.


DNA polymerase of Dictyostelium discoideum.

Only one molecular weight species of DNA polymerase was found in different developmental stages of the eukaryotic microorganism Dictyostelium discoideum. The molecular weight of this DNA polymerase is estimated to be about 127 000 by sucrose gradient centrifugation. The enzyme is present in all stages of growth and development, including dormant spores. All DNA polymerase activity is lost upon incubation of the crude extract with N-ethylmaleimide. The reaction properties, molecular weight and N-ethylmaleimide sensitivity of the D. discoideum DNA polymerase are similar to those of the DNA polymerase-alpha from mammalian sources.


Interaction of phosphatidylcholine with bovine serum albumin. Specificity and properties of the complexes.

Phosphatidylcholine dispersed on Celite was rapidly solubilized by neutral bovine serum albumin solutions. Stable protein-lipid complexes were isolated by Agrose gel filtration or by ultracentrifugal flotation in high density solvents, and the physicochemical properties of the complexes were investigated in terms of the stoichiometry of binding, effect of fatty acid ligands on phosphatidylcholine binding, effect of high ionic strength on the stability of the complexes, intrinsic fluorescence and circular dichroism spectra, and sedimentation velocity coefficients. Complexes containing from 2 to 30 phosphatidylcholine molecules per protein molecule were observed; however, no saturation of binding sites could be detected in this range of molar ratios. Oleic acid binding by serum albumin prevents interaction of the protein with phosphatidylcholine, indicating possible competition of these ligands at low contents of the phospholipid. For molar ratios of up to 10 phosphatidylcholine molecules per serum albumin, binding is primarily due to hydrophobic interactions that have no effect on the overall shape and secondary structure of the native protein except for local modifications at tryptophan residues, whose fluorescence becomes quenched and blue shifted on phosphatidylcholine binding. Similar phosphatidylcholine uptake experiments performed with a series of globular proteins indicated that the lipid extraction from Celite surfaces is a non-specific process, accelerated by several other proteins (e.g. aldolase, egg albumin, chymotrypsinogen, soybean trypsin inhibitor, and the major apolipoprotein from bovine serum high density lipoprotein). Formation of stable protein-lipid complexes, however, was only observed with bovine serum albumin, which in contrast to the other proteins is known to have affinity binding sites for anions with hydrophobic side chains.


Distribution of 8 trace metals in beef heart tissue.

The concentrations of 8 trace metals in 15 discrete anatomic sites of 11 beef hearts have been determined by emission spectrometry. The metals determined were copper, aluminum, barium cesium, tin, strontium, lead, and molybdenum. Anatomic sites samples included aorta, main pulmonary artery, tricuspid valve, mitral valve, right and left coronary arteries, os cordis, right atrium, left atrial appendage, right and left ventricles (free wall), left ventricle-papillary muscle, interventricular septum, crista supraventricularis, and left bundle branch. Statistical analysis of the data incorporated a multivariate analysis of variance and a Duncan analysis. The results showed that many tissues of the beef cardiovascular system have distinctive trace metal compositions.


Self-association of beta-lactoglobulin c in acetate buffers.

The self-association of beta-lactoglobulin C at pH 4.65 (23 degrees C) in acetate buffer has been studied at various temperatures, 10, 16, 20 and 25 degrees C, by a series of sedimentation equilibrium experiments. Two different buffers were used. Buffer I with an ionic strength of 0.1 consisted of 0.1M acetic acid and 0.1M sodium acetate; buffer II had 0.1M KCl in addiation so that its ionic strength was 0.2. The variation of the apparent weight average molecular weight, Mwa, with the total solute concentration, c, was characteristic of a self-association. In contrast to the behavior of beta-lactoglobulin A in acetate buffer, the association of beta-lactoglobulin C did not proceed beyond dimer. Furthermore, within the experimental error, the self-association of beta-lactoglobulin C was independent of temperature and ionic strength; all experimental data could be put on the same Mwa (or M1/Mwa) vs. c plot! Several models were used to test the self-association, and a monomer--dimer association with K2 = 2.10 X 10(3) dl/g and BM1 =- 1.2 X 10(-2) dl/g seemed to give a good description of the M1/Mwa vs. c curve.


Hormonal changes and mood in the puerperium.

This investigation is an attempt to test the common supposition that postpartum emotional disturbance is related to hormone changes. A group of 27 normal pregnant women were assessed three times before delivery and sixteen times in the six weeks following delivery. During the first two interviews baseline data on personality and other personal variables were obtained. On each occasion blood was taken and three measures of clinical status and mood were completed. Plasma LH, FSH, total oestrogen and progesterone results are presented in detail and the results of prolactin assays mentioned more briefly. An attempt to correlate hormone findings and clinical findings is described. This failed to produce any strong evidence that hormones are related to mood at this time, although hormone changes were correlated weakly with a few specific symptoms. Some of the unexpected clinical findings and technical difficulties of the study are discussed, with special reference to possible further research in this area.


Obstetric practice and outcome of pregnancy in Cardiff residents 1965-73.

Trends in management and outcome of pregnancy in Cardiff residents from 1965 to 1973 were reviewed. The mean age and parity of parturients fell. Hospital delivery became almost universal, monitoring the fetus during pregnancy was introduced, and induction and acceleration of labour became commonplace. These developments were not associated with any striking change in either the total perinatal death rate or the timing or cause of perinatal death. Possibly a real change in perinatal mortality between 1965 and 1973 was masked by random fluctuation of small numbers, or possibly factors peculiar to the Cardiff population prevented a decrease in  perinatal mortality that would otherwise have resulted from improved medical care. Only by large-scale randomised trials can the true value of induction and other medical developments be assessed.


Obstetric practice in the Oxford Record Linkage Study Area 1965-72.

The secular trends in induction in 111 818 births over eight years in the largely rural areas of Oxfordshire and west Berkshire were analysed. Although the induction rate started to rise only in 1969, the forceps and episiotomy rates had been increasing throughout, but both procedures were always twice as prevalent in induced as in non-induced cases. The proportion of women given an anaesthetic was also consistently higher in induced cases, but that of women with long labours fell considerably over the period. The reduction in stillbirth rate was more apparent in induced than in non-induced births.


Effect of 5-bromo-2'-deoxyuridine or cytosine-B-D-arabinofuranoside hydrochloride on myelination in newborn rat cerebellum cultures following removal of myelination inhibiting antiserum to whole cord or cerebroside.

Myelination was inhibited in cultures of newborn rat cerebellum by exposure to antisera prepared by injecting rabbits with whole guinea pig spinal cord or cerebrosixde mixed with bovine serum albumin. At 15 days in vitro (DIV), when 90-100% of our control cultures were myelinated, antiserum inhibited cultures were washed and refed routine culture medium or medium containing 5-bromo-2'-deoxyuridine (BUdR) or cytosine-B-D-arabinofuranoside hydrochloride (ara-C) in concentrations previously shown to inhibit myelination in our system. The disinhibited cultures myelinated within 2-5 days of removal of antiserum regardless of the presence of BUdR or ara-C. The evidence suggests that the myelin-forming oligodendrocytes do not undergo cell division before myelination during disinhibition from antiserum. In addition, cultures exposed to BUdR on 5-7 DIV in the presence of antiserum and disinhibited at 10 DIV did not have myelinated axons as observed by light microscopy at 15 DIV. Thus, the BUdR sensitive oligodendrocyte division remained intact in antiserum inhibited cultures. We conclude that the antisera inhibit myelin formation without interrupting the pattern of cell division or the covert differentiation of oligodendrocytes.


The influence of lighting conditions upon the level and course of increase in specific activity of serotonin N-acetyltransferase in the developing chick pineal gland.

A diurnal cycle in level of serotonin acetyltransferase (acetyl-C0A:arylamine N-acety-transferase (EC 2.3.1.5) activity was found in the pineal gland of chicks aged 16 to 20 days maintained under lighting conditions. Diurnal variation in levels of activity was markedly reduced in the pineal gland of birds kept in constant darkness, and suppressed in the gland of chicks under constant illumination. High levels of activity attained during the dark phase of the normal cycle rapidly declined when the birds were transferred to the light. The light phase level of serotonin acetyltransferase of the pineal increased progressively from the 11th day of incubation to about 1 week post-hatch. This course of increase in enzyme activity was largely unaffected by lightning conditions. Under conditions for assay of serotonin acetyltransferase activity in the chick pineal gland and brain, radioactive serotonin gave rise to N-acetylserotonin, 5-hydroxy-indoleacetic acid and a further unidentified metabolite, which was quantitatively the major product.


Studies on the cell wall of Schizophyllum commune. Permethylation and enzymic hydrolysis.

Two polysaccharide fractions S-glucan were isolated from the cell walls of Schizophyllumcommune. S-Glucan is primarily a linear 1,3-alpha-glucan with occasional 1,6-alphalinkages as shown by permethylation and partial acid hydrolysis. The glucan fraction also contains a small amount of xylose. The R-glucan fraction is a mixture of two polysaccharides, chitin and a highly branched glucan with linear 1,6-beta and 1,3-beta segments and 1,6-beta branching. This conclusion is based on permethylation studies and enzymic hydrolyses.


Surface polypeptides of the Chinese hamster ovary cell; an immunochemical study.

Antibodies elicited by the injection of live Chinese hamster ovary cells (CHO) into rabbits precipitated four major components from detergent extracts of CHO membranes. The four components, of molecular weights 200000, 125000, 95000 and 41000 daltons, corresponded to cell surface components identified by the lactoperoxide surface label technique.


[Fate of the two largest ovarian follicles of sheep following destruction of the largest follicle and/or the removal of the corpus luteum at two times in the cycle].

Marking of the largest (F1) and the second largest (F2) follicles of the ovaries of the sheep with India ink has allowed us to observe the fate of these follicles after different treatments. All the F1 and F2 follicles of the controls marked on the 10th day of the cycle became atretic, but when the marking was done on the 14th day, only the F1 (of the marked follicles) ovulated. Destruction of the F1 with or without removal of the corporalutea allowed some of the F2 to ovulate, wheras only removal of the corpora lutea decreased the number of F1 ovulating. It thus appears that the fate of the F1 is related to the time of the cycle and that the F1 could have an inhibitory effect on the ovulation of the F2. Removal of the corpora lutea also induced the development of several cystic follicles and decreased the total ovulation rate.


Products of anaerobic phloroglucinol degradation by Coprococcus sp. Pe15.

Under anaerobic conditions, resting cell suspensions of Coprococcus sp. Pe15 degraded 1 molecule of phloroglucinol to 2 molecules of acetic acid and 2 molecules of carbon dioxide. The organism metabolized the flavonoids rhamnetin and quercetin anaerobically in 20% rumen fluid medium but failed to grow under similar conditions at the expense of any of 39 other aromatic or flavonoid compounds tested.


An electron-microscopic study of Herpetomonase sp. (Leptomonas pessoai).

The fine structure of the promastigotes of Herpetomonas sp. (Leptomonas pessoai) kept in a defined medium at 28 degrees C is described. This portozoon reveals several features in common with other trypanosomatids. A membrane-bounded organelle measuring 0.2 to 0.8 mum in diameter, similar to that described as peroxisome in Crithidia fasciculata, was also observed. A large cavity, located between the nucleus and the kinetoplast and containing vesicles and small particulate material is discussed in this paper.


Some chemical and structural features of the conidial wall of Trichoderma viride.

Cell wall of spores of Trichoderma viride contains polymers similar to those of mycelial cell wall, such as beta-(1 leads to 3), beta-(1 leads to 6) glucans and protein, but chitin, always present in the mycelium, cannot be found in spores. Melanin, which in other fungi appears associated with chitin, replaces this polymer in the spore wall of T. viride and is located in the outermost layer. Attempts to characterize the pigment of the spore wall indicate that it is a non-indolic melanin-like polyphenol.


Heterokaryosis and parasexuality in Pyricularia oryzae Cavara.

Auxotrophs in three different races of Pyricularia oryzae were obtained following ultraviolet mutagenesis, and tested for complementarity on minimal medium. Prototrophic growth resulted between combinations of different auxotrophs of single strains, but not between strains. The growth did not result form crossfeeding, but required at least transitory heterokaryosis. Putative diploids, i.e., large prototrophic conidia, were readily isolated from the heterokaryons. In addition, conidia with nearly every possible combination of parental markers were found, showing that P. oryzae does indeed complete the parasexual cycle. The degree of genetic recombination found here may help to explain the extreme variability of this organism.


Isolation of ioxynil degraders from soil-enrichment cultures.

A soil enrichment technique was used to isolate microorganisms which could degrade ioxynil (3,5-diiodo-4-hydroxybenzonitrile). Many isolates obtained were able to degrade ioxynil to various products. However, only a fungal isolate (Fusarium solani) and a Gram-negative bacterium (Klebsiella ozaenae) released 14CO2 from ring-labeled ioxynil. No appreciable degradation was detected in pure cultures without the addition of exogenous nutrients. Results indicated that the degradation of ioxynil to CO2 proceeded more slowly in pure culture. Ioxynil was degraded in pure culture at a faster rate by F. solani than by K. ozaenae. Analyses of radioactivity distribution in the cultures indicated that a sizable fraction of radioactivity was in the form of polar products. Several degradation products were detected in the ethyl acetate extracts by thin-layer chromatography and subsequent radioautography. Screening of pure cultures of ioxynil degraders revealed that most isolates degraded ioxynil to the same products which were extractable with ethyl acetate.


The development in beef cattle manure of Petriedllidium boydii (Shear) Malloch, a potential pathogen for man and cattle.

Petriellidium boydii (Allescheria boydii) dominated the mycoflora of manure samples form three beef cattle feedlots after incubation at room temperature for 4 months. The possible dangers associated with this pathogenic fungus, which causes mycotic abortion in livestock, pulmonary allescheriasis in man, and mycetomas in both man and animals, are discussed. This fungus could create a health hazard in feedlots where in situ manure decompostion is encouraged.


Idiopathic thrombocytopenic purpura in lymphoma.

Five patients with idiopathic thrombocytopenic purpura (ITP) associated with malignant lymphoma are presented, and twelve other cases reported in the literature are reviewed. Thirteen patients (76%) had Hodgkin's disease; two had diffuse histiocytic, one, diffuse mixed lymphoma. Idiopathic thrombocytopenic purpura presented as the sole clinical manifestation of initial or recurrent lymphoma in 10 of 15 cases (67%). The spleen was not palpable even in patients who showed involvement with lymphoma at splenectomy. Three patients were asplenic at the onset of idiopathic thrombocytopenic purpura. Successful therapy of the thrombocytopenia correlated with effective control of the underlying lymphoma. Splenectomy gave favorable results in 9 of 10 cases (90%) and uncovered occult lymphoma in five patients. Chemotherapeutic drugs were successful in seven of 12 trials (58%) and only when the underlying disease was also controlled. The overall prognosis was related to the status of the lymphoma and was not altered by the occurrence of the idiopathic thrombocytopenic purpura.


In vitro cultivation of epithelial cells derived from tumors of the human urinary tract.

Finely minced explants from 54 TCC2 of the human urinary tract were cultured in vitro in an attempt to establish cell lines. Cells with epithelial morphology grew out from 48 tumor explants, and long-term cell cultures were established from 10. Six of the cell cultures have been maintained for over 18 months with 50 to 70 transfers and, therefore, are considered permanent cell lines. The epithelial cells in the established cultures are small, exhibit rapid doubling time, and show multilayering. The cells were examined both microscopically and by cultivation techniques, and they were found to be free from contaminating microorganisms, including Mycoplasma. The established cultures grow rapidly in roller bottles and, therefore, can be produced in large quantities. These cells also remain viable after being stored for 3 years in liquid nitrogen.


Control of local and regional subclinical disease by radiation therapy.

An irradiation dose of 5000 rads in 5 weeks appears to be highly efficient for eradication of subclinical disease in the oropharynx or nasopharnyx. Irradiation alone in the range of 4500 to 5000 rads, five treatments per week, 4 1/2 to 5 weeks, appears to be greater than 90% efficient for eradicating subclinical disease for both squamous-cell carcinoma and adenocarcinoma in regional lymph nodes. The site of origin of the primary squamous-cell carcinoma or adenocarcinoma seems to be independent of the response in the regional lymph nodes. It is unknown whether concomitant elective chemotherapy will modify the dose for control required for subclinical disease in squamous-cell carcinoma or adenocarcinoma.


Potentiation by guanine nucleosides of the growth-inhibitory effects of adenosine analogs on L1210 and sarcoma 180 cells in culture.

The growth-inhibitory effect of 6-methylmercaptopurine riboside (MMPR) against leukemia L1210 cells in culture was dramatically potentiated by the addition of guanine nucleosides to the medium. In the presence of either deoxyguanosine or guanosine, the concentration of MMPR that caused 50% inhibition of growth was 35 times lower than in the absence of these nucleosides. Similar potentiation was also observed against Sarcoma 180 cells in culture by guanosine. The metabolic basis of this synergism was approached in a study of the incorporation of [14C]glycine into 5'-phosphoribosyl-N-formylglycinamide in Sarcoma 180 cells. The results show that the site of inhibition resulting in synergism is an early step in purine biosynthesis, probably phosphoribosyl pyrophosphate amidotransferase (EC 2.4.2.14). In the L1210 cell system, the addition of hypoxanthine to the medium prevented the potentiation of MMPR by guanine nucleosides supporting the conclusion that the site of the synergistic interaction involves purine biosynthesis de novo. While hypoxanthine partially reversed the growth-inhibitory effects of MMPR, an even higher degree of protection was observed in the presence of both uridine and hypoxanthine, suggesting that MMPR may have additional sites of action concerned with pyrimidine metabolism.


Common and individually specific chromosomal characteristics of cultured human melanoma.

Since individual chromosomes can be accurately identified by new banding techniques, atebrin fluorescence was used for chromosome analysis in six cell lines and two primary outgrowths derived from human malignant melanoma. Gross aneuploidy was seen in all specimens, but each culture contained at least 1 distinctive marker chromosome specific for that cell line in 87 to 100% of metaphases. One of the primary explants contained a marker that was demonstrable in fresh tissue and persisted through 2 weeks of culture. The same marker was found in all metaphases from 2 different metastases, but skin fibroblasts from the same patient had a normal chromosome complement. No common marker for human melanoma was found, but in 6 of the 8 cultures the most frequently found marker was formed by a brightly banded chromatid addition. Relative polysomy for Chromosome 7 was found in 7 of the 8 cultures and, for Chromosome 22, in 8 of the 8 cultures. The frequency of polysomy of Chromosomes 7 and 22 was significant at the 5% level.


Separation of cells with histochemically demonstrable acid phosphatase activity from suspensions of cells from human prostatic carcinomas in an isokinetic gradient of Ficoll in tissue culture medium.

This report describes the separation of cells exhibiting histochemically demonstrable acid phosphatase from suspensions of cells obtained from human prostatic carcinomas by velocity sedimentation. In the unseparated suspensions of cells, 40.5 +/- 7.7% of nucleated cells contained histochemically evident acid phosphatase. After cells were separated by velocity sedimentation, 86.4 +/- 9.4% of the nucleated cells in the purest fractions exhibited histochemically demonstrable acid phosphatase activity. More than 95% of these cells excluded trypan blue. To our knowledge, this is the first report of a method for the separation of viable epithelial cells from human prostatic carcinomas.


A characterization of the immunosuppressive activity of adriamycin and daunomycin on humoral antibody production and tumor allograft rejection.

The effects of adriamycin (AM) and its analog daunomycin (DM) on immunological responsiveness have been investigated in an effort to elucidate whether a differential interaction of the two drugs with the immune system could play a role in the higher antineoplastic activity of AM. It was found that AM induced a greater reduction in the number of antibody-producing cells after primary stimulation with sheep erythrocytes, whereas DM was more suppressive on the secondary response to the same antigen. Primary reactivity to the T-independent antigen S-III was reduced by AM, whereas DM was ineffective in the same conditions even at high doses. In addition, when a tumor allograft model was investigated, DM was significantly more immunosuppressive than was AM administered at equitoxic doses. In contrast, these agents displayed similar activity in reducing bone marrow stem cells and in inhibiting DNA synthesis in this organ. The possibility that the different immunosuppressive capacity of AM and DM contributes to the greater antitumoral activity of the former is advanced.


Uptake of partially thiolated DNA by ascites tumor cells.

The uptake and intracellular localization by Ehrlich ascites cells of partially [35S]thiolated homologous DNA ("antitemplate") were studied in comparison with that of the corresponding unmodified [3H]DNA, at 37 degrees and 0 degrees, under standardized conditions. For the unmodified DNA, washing the cells after incubation with 0.08 M iodoacetate (in 0.15 M NaCl) alone gave high but reproducible uptake values (23%); washing with 1 M NaCl reduced the cell-associated DNA to 12% (less than 1% at 0 degrees). It appears that 1 M NaCl is able to remove DNA reversibly bound to the cells, similarly to DNase treatment. Approximately 5% of the input [3H]DNA was taken up into the cell nuclei. Diethylaminoethyl dextran (1:1, by weight) greatly enhanced the cellular uptake of [3H]DNA. In the case of [35S]thiolated DNA, the rate as well as the extent of uptake was significantly higher (33%). Washing the cells with 1 M NaCl or treatment with DNase caused relatively small decrease in the total cell-associated [35S]thiolated DNA, the bulk of which (22% of input) was recovered in the isolated nuclei. Stimulation by diethylaminoethyl dextran of the uptake of [35S]thiolated DNA could not be established because of the insolubility of the 1:1 complex in 1 M NaCl. Excess calcium ions during incubation dramatically increased the uptake of the thiolated DNA at 37 degrees (but not at 0 degrees) by the cells (to 90 to 100%) and into the nuclear fraction (to 70% of the total [35S]DNA input). The calcium salt procedure appears to be applicable to the in vivo testing of thiolated DNA's as potential chemotherapeutic agents.


Enhancement of 9-beta-d-arabinofuranosyladenine cytotoxicity to mouse leukemia L1210 in vitro by 2'-deoxycoformycin.

2'-Deoxycoformycin (2'-dCF), a potent inhibitor of adenosine deaminase, was tested in combination with 9-beta-D-arabinofuranosyladenine (ara-A) and 9-beta-D-arabinofuranosyladenine 5'-formate for cytotoxic activity against mouse leukemia L1210 in culture. 2'-dCF, which alone had no activity, significantly enhanced cytostatic and cytotoxic activities of ara-A and its more soluble derivative, 9-beta-D-arabinofuranosyladenine 5'-formate; the latter 2 agents, when tested at equimolar concentrations, were equivalent in their effects on proliferation and viability. The therapeutic response of mice bearing the in vitro line of L1210 cells (L1210/C2) to combination therapy with 2'-dCF and 9-beta-D-arabinofuranosyladenine 5'-phosphate was comparable to that reported elsewhere for therapy of mice bearing the parent in vivo line. Continuous exposure of cultured L1210 cells to ara-A and 2'-dCF induced a prolonged period of unbalanced growth, characterized by inhibition of proliferation and DNA synthesis while RNA and protein synthesis continued; exposure periods in excess of a single population doubling were required to achieve significant cell kill. Potentiation of ara-A activity against the relatively insensitive mouse leukemia L1210 was attributed to increased stability of ara-A resulting from 2'-dCF inhibition of adenosine deaminase.


An extracellular fungal polysaccharide composed of 2-acetamido-2-deoxy-D-glucuronic acid residues.

The black yeast-like fungus NRRL YB-4163, now tentatively identified as Rhinocladiella elatior Mangenot, has been found to produce an extracellular microbial polysaccharide composed mainly of 2-acetamido-2-deoxy-D-glucuronic acid residues. Polysaccharide (PS) YB-4163, when isolated in good yield as the neutral potassium salt, dissolves readily in water to produce extremely viscous solutions, which form stable foams and emulsions. By depolymerizing PS YB-4163 with [14C]methanol-HCl, the polysaccharide can be both identified and quantitated radiochemically by determining the individual [14C]methyl glycosides after their separation by paper chromatography. When the methyl glycosides of PS YB-4163 were reduced with NaB3H4, only the methyl glycosides of 2-acetamido-2-deoxy-D-[6-3H]glucose were found. Analysis of the monosaccharide released from carboxyl-reduced PS YB-4163 by acid hydrolysis or methanolysis also showed 2-acetamido-2-deoxy-D-glucuronic acid to be the main constituent. Previously, the only polysaccharides known to be composed entirely or hexosaminuronic acid have been cellular products from pathogens. Of these, the antigenic polysaccharide (SPSA) from Staphylococcus aureus is composed entirely of 2-amino-2-deoxy-D-glucuronic acid, but its amino groups are substituted equally with acetyl and N-acetylalanyl groups. The specific optical rotation of PS YB-4163, [alpha]20D -75 degrees (c 0.5, water), is similar to that of SPSA (-91 degrees), and suggests beta-D-linkages that must be either (1 leads to 3) or (1 leads to 4).


Fetal hemopoiesis in diffusion chamber cultures. I. The pattern of pluripotent stem cell growth.

The growth pattern of fetal liver (FL), normal adult bone marrow (NABM) and regenerating (post Velban treatment) adult bone marrow (RABM) colony forming units (CFU) cultured in diffusion chambers (DC) was studied. When twenty CFU were implanted into DC the recovery of CFU after 4 days with FL, NABM or RABM was 133 +/- 7, 19 +/- 2 and 34 +/- 2 CFU, respectively. The transplantation fraction of CFU from NABM decreased from 10-4% on day 0 to 6-9% on day 4; that of FL did not change from the initial 6-2%. The growth rate of CFU derived from FL was substantially greater than that from NABM. The relative growth of FL and RABM CFU was clearly inhibited when the concentration of cells cultured was increased. Spleen colonies from FL cells before culture were larger (P less than 0-005) than colonies from NABM but after 7 days of culture there was no difference between the two groups. Histological examination of spleen colonies showed that after DC culture FL and NABM CFU were differentiating along the three normal pathways. These data suggest that intrinsic differences exist between fetal and adult stem cells in the in vivo diffusion chamber culture system.


Hybridization of a human myeloma permanent cell line with mouse cells.

A population of hybrid cells derived from the fusion of a permanent human myeloma cell line, which secretes complete IgE, and a subline of mouse L cells, did not secrete IgE as evidenced by sensitive immunosorbent tests. Also, the hybrid cells were observed not to contain intracellular IgE (epsilon or lambda chains) in amounts to be detectable by fluorescent antibody techniques. The doubling times and cell cycle parameters of the hybrid cells were found to be similar to those of the slow-growing parental human myeloma cells, in addition, the growth of the hybrid cells was characterized by a higher degree of contact inhibition than the parent mouse cells.


The fine structure of the hypothalamic secretory neurons of the white-crowned sparrow, Zonotrichia leucophrys gambelii (Passeriformes: Fringillidae). II. Magnocellular and parvocellular nuclei of the rostral hypothalamus.

The fine structures of the neurons and neuropils of the magnocellular supraoptic nucleus and the parvocellular nuclei of the rostral hypothalamus, including the suprachiasmatic and medial, lateral and periventricular preoptic nuclei, and the neuronal apparatus of the organum vasculosum laminae terminalis, have been examined in the male White-crowned Sparrow, Zonotrichia leucophrys gambelii, by correlated light and electron microscopy. The magnocellular supraoptic nucleus is characterized by large neurosecretory perikarya which contain a well developed Golgi complex and dense-cored granules 1,500-2,200 A in diameter. The neuropil displays axons, dendrites and glial fibers. Some axonal profiles contain dense-cored vesicles 800-1,000 A in diameter and clear vesicles 500 A in diameter. Axo-somatic and axo-dendritic synapses are conspicuous in this nuclear region. The suprachiasmatic nucleus is characterized by an accumulation of small neurons with moderately developed cellular organelles and some dense-cored granules, approximately 1,000 A in diameter. The profiles of axons within the neuropil contain dense-cored granules 800-1,000 A in diameter and clear vesicles 500 A in diameter. The neurons of the medial preoptic nucleus are relatively large and exhibit well developed cellular organelles and dense-cored granules 1,300 to 1,500 A in diameter. Granular materials are formed within the Golgi complex. The medial preoptic nucleus is rich in secretory perikarya. Occasionally, neurons with granules 1,500-2,200 A in diameter are encountered in the lateral preoptic and periventricular preoptic nuclei. They may be considered as scattered elements of the magnocellular (supraoptic and paraventricular) system. The organum vasculosum laminae terminalis consists of three layers, i.e., ependymal, internal and external zones, and exhibits a vascular arrangement similar to that of the median eminence. The perikarya of the parvocellular neurons and their axons in the internal zone contain numerous secretory granules ranging from 1,300 to 1,500 A in diameter.


Ribosomes from Xenopus laevis eggs and embryos in a cell-free protein-synthesizing system: translational regulation.

Three types of ribosomal preparations from Xenopus laevis eggs and embryos were tested in a cell-free system to study possible translational regulation of protein synthesis as mediated by the ribosome during early amphibian development: type 1, a crude high-speed sediment, mainly containing monoribosomes completely dissociable by 0.5 M KC1; type II, ribosomes washed with 0.5 M KC1; and type III, ribosomes treated with puromycin - 0.5 M KC1. All three types showed an active response to the addition of poly[U]. Type III was found to be the most active: levels of incorporation of 30 phenylalanine residues/ribosome were reached. In all three cases ribosomes prepared from unfertilized eggs were 30-40% less active in vitro than those from cleavage and gastrula stages.


Amphibian embryo protease inhibitor. III. Binding studies on the trypsin inhibitor and properties of its yolk-bound form.

Amphibian embryo trypsin inhibitor (ATI) is shown to be a temporary inhibitor in which full trypsin activity can eventually be restored. Lineweaver-Burk plots and other binding data indicate a reversible change from competitive to non-competitive inhibition during incubation of enzyme with inhibitor in the absence of substrate. The inhibitor is active while sequestered in the yolk, and the sequestered inhibitor co-purifies with lipovitellin when the yolk is fractionated. It is further demonstrated that the inhibitor slowly leaches from the yolk in a soluble form, and it is suggested that this leakage is the result of a negative feedback controlled proteolysis within the yolk. Possible interactions between ATI and endogenous trypsin-like proteases are discussed with particular regard to the potential involvement of those materials in the control of certain developmental events; notably melanogenesis.


Studies on the mechanism of in vitro estradiol-17 beta induced synthesis of phosvitin in chick embryo liver cells.

The effect of estradiol-17-beta treatment on phosvitin synthesis by cultured chick embryo liver cells has been studied. Phosvitin synthesis occurs approximately 15 hr of hormone treatment; the synthesis being blocked by actinomycin D treatment suggests that RNA synthesis is required. The life time of the newly synthesized RNA is at least 24 hr. The significance of these findings with respect to the mechanisms involved in hormone-mediated protein synthesis is discussed.


Growth response to lectins in chick embryo cells at different stages of development.

We report here the effect of Robinia lectin and Concanavalin A on fibroblasts and liver cells from chick embryos between the 8th and the 20th day of development. This was observed in vitro after different times of cultivation. There is evidence that these lectins decrease cell number in cultures from young embryo cells but that they stimulate the proliferation of older embryo cells. The optimum concentration of either lectin was 3 mug/ml. No effect was observed on 12- and 14-day cells at the different concentrations of lectin used. The agglutination of fibroblasts by these lectins regularly decreased from the 8th to the 16th day of development. Liver cells however were agglutinated at no stage. These results could perhaps be explained in terms of cell surface changes either during the course of ontogeny or as a result of lectin treatment.


Parachute accessory anterior mitral valve leaflet causing left ventricular outflow tract obstruction. Report of a case with emphasis on the echocardiographic findings.

A case of left ventricular outflow tract obstruction caused by a parachute accessory anterior mitral valve leaflet is presented. The abnormality was detected in an asymptomatic patient by systolic murmur and thrill and was demonstrated by echocardiography, as well as by cardiac catheterization and surgery. These findings are presented. Several previously reported cases of a similar nature are reviewed. An identical case was not found in the English literature.


Calculation of estrogen excretion rates from urinary estrogen to creatinine ratios.

1. The excretion of estrogens by 42 menstruant and 7 pregnant (1st trimester) women ranged from 2 - 550 mug/24hr. 2. These changes were followed by measuring estrogen to creatinine ratios in small samples of urine. 3. Values of the ratio changed little in each 24 hr, and correlated closely with daily estrogen output (r = 0.971; P less than 0.001; estrogens (mug/24hr) =(see article. 4. The data suggests that ovarian activity in women, including those given gonadotropins to induce ovulation, may be followed by measuring estrogen to creatinine ratios in small samples of urine; the collection of 24 hr samples is unnecessary.


Rapid radioimmunoassay of total urinary estriol.

We describe a radioimmunoassay for total urinary estriol in pregnancy. A 25-mul aliquot of the urine specimen is acid hydrolyzed, neutralized, and diluted before assay. We use rabbit antisera against estriol-6-(O-carboxymethyl)oxime/bovine serum albumin and ammonium sulfate precipitation at room temperature. Results are unaffected by glucose or methenamine mandelate, a urinary tract antiseptic. Using semi-automatic pipetting equipment, one laboratory technologist can complete 50 assays within 8 h. This technique is both reliable and convenient and should decrease the expense of routine estriol assays.


Radioimmunoassay of iodinated gentamicin.

A radioimmunoassay is described for the aminoglycoside antibiotic, gentamicin. Iodinated gentamicin was prepared using an 125I-labeled acylating agent: 3-(4-hydroxyphenyl)propionic acid N-hydroxysuccinimide ester which spontaneously reacts with gentamicin to produce an iodinated conjugate. The radioimmunoassay with iodinated gentamicin produced a straight line on a logit/log plot yielding an assay sensitivity of 80 pg. A Scatchard plot plus an equilibrium technique were used to estimate the affinity constant at 8 X 10(9) 1 mol-1 and 10.5 X 10(9) 1 mol-1, respectively. Kanamycin and neomycin, two chemically related aminoglycoside antibiotics, did not crossreact with gentamicin until concentrations were 10(5) greater than those found in blood. Assay of patients' serum samples by both a microbial assay and radioimmunoassay resulted in a statistically significant (p less than 0.001) correlation between the two methods.


Specific precipitating antibody to human urinary erythropoietin.

In the present experiments, human urinary erythropoietin was purified from the urine of a patient suffering from pure red cell aplasia, using ethanol precipitation, gel filtration on the Sephadex G-100 column and electrofocusing. The anti-human urinary erythropoietin was prepared by injecting the isolated erythropoietin with Freund's complete adjuvant into rabbits intracutaneously. The specificity of the anti-human urinary erythropoietin was ascertained by a double diffusion technique and immunoelectrophoresis, observing a single precipitation line at the alpha2-globulin region against human serum or human urinary erythropoietin (Standard B). This precipitating anti-human urinary erythropoietin inhibited completely the effects of erythropoietin (Standard B) on the heme synthesis of bone marrow cells in vitro. The precipitating antigen with the specific anti-human urinary erythropoietin, that is, erythropoietic stimulating factor of the fraction 2, had the ability to enhance heme synthesis in bone marrow cells in vitro. The specific anti-human urinary erythropoietin makes it possible to determine the serum erythropoietin levels in various diseases by means of quantitative immunoelectrophoresis.


Retinol binding protein in serum and in urine of glomerular and tubular nephropathies.

RBP behavior was studied in different kidney diseases. Serum RBP was increased in chronic renal diseases when the serum creatinine increased. RBP clearance and RBP clearance as a percentage of albumin clearance increased when GFR decreased; this is more evident in tubular kidney diseases. Urinary RBP loss as a percentage of urinary albumin loss increased especially in kidney diseases with persistent tubular lesions.


Air embolus following pulmonary interstitial emphysema in hyaline membrane disease.

Two cases of hyaline membrane disease are presented complicated by severe, interstitial emphysema and resulting in death from air embolus. This is consistent with the experimental evidence of other investigators that shows that such an air embolus may be more common complication of hyaline membrane disease than previously expected.


Tetralogy of Fallot with discrete fibrous subaortic stenosis.

This is the first reported case of discrete fibrous subaortic stenosis in tetralogy of Fallot. Transaortic excision of the subaortic membrane was readily accomplished. To detect this and other forms of left ventricular outflow-tract obstruction in tetralogy of Fallot before surgery, routine selective left ventriculography is advocated.


Parameters of spikes in human epilepsy.

The parameters of 600 spikes were studied in 100 epileptic patients. Spikes were subdivided morphologically into: monophasic spikes, diphasic spikes, spike-slow wave complexes, polyspike-slow wave complexes and spike followed by spike. The following parameters were analyzed: polarity, amplitude, duration and sequence, 88% of spikes were negative. 98% of spikes raised 30% above the background activity. Spike duration varied, the shortest spike had a 9 msec duration. The range of duration varied from 9 to 200 msec with a mean of 45.06. 75% of spikes were followed by a deflection lasting from 130 to 200 msec. No evidence was found in our study for the differentiation of sharp waves from spikes.


Termination at midpregnancy of the two daily surges of plasma prolactin initiated by mating in the rat.

The pattern of plasma prolactin following uterine cervical stimulation consists of two surges each day, one nocturnal, occurring between 0100-0900 h (lights on 0600-1800 h), and one diurnal, occurring between 1500-2100 h. This pattern of prolactin continued throughout pseudopregnancy; the last surge was observed on the morning of day 11 (day 1 was taken as the first day of diestrus of pregnancy or pseudopregnancy). Prolactin levels remained low thereafter until the spontaneous proestrous surge on the afternoon of day 12, signalling the onset of a new estrous cycle. In contrast, the two daily prolactin surges did not continue throughout pregnancy, and in fact, were terminated sooner in pregnant animals than in pseudopregnant animals. The last diurnal surge was observed on day 8 while the last nocturnal surge was observed on day 10. The early termination of prolactin surges during pregnancy correlated with the increased secretion of rat placental lactogen. However, placental extracts obtained from day 11 of pregnancy and injected in large doses failed to inhibit prolactin surges in pseudopregnant animals. Prolactin surges also continued for a longer period of time in pseudopregnant rats bearing decidualized uteri than in pregnant animals. Thus, the two major components of pregnancy that differed from pseudopregnancy, that is, the presence of rat placental lactogen or decidual tissue, did not appear to account for the early termination of prolactin surges during pregnancy.


Hypothalamic and suprachiasmatic uptake of serotonin in vitro: twenty-four-hour changes in male and proestrous female rats.

Twenty-four-hour changes in the in vitro serotonin (5-HT) uptake capacity of hypothalamic homogenates and of "Vibratome" slices of the suprachiasmatic nuclear region (SNR) of the hypothalamus were studied in 60-90-day-old Holtzman (albino) rats. Animals acclimated to artificially illuminated (fluorescent, LD 12:12) and temperature controlled (22 +/- 2 C) rooms were killed 6% each of 8 time points. Synaptosomal fractions from homogenates of whole hypothalamus, and slices of the SNR were incubated for 20 min in Krebs-Henseleit buffer with [3H]5-HT. Males showed a single daily peak in SNR uptake at the start of darkness, and a minimum near the onset of light, while a more complex pattern containing 3 peaks and minima occurred in uptake by hypothalamic homogenates. Proestrous females showed a single high amplitude peak SNR uptake during the critical period, just prior to the plasma LH peak determined in the same animals by radioimmunoassay. It is suggested that this short-term and 4-fold increase in SNR uptake of 5-HT may serve to limit free 5-HT and its inhibitory or other effects on the gonadotropin release hormone system and thereby on LH release and ovulation.


Synthesis of ribosomal RNA during the mitotic cycle on the slime mould Physarum polycephalum.

1. An isotope dilution technique has been used to analyze the synthesis of metabolically stable nucleic acids during the mitotic cycle in surface plasmodia of the slime mould Physarum polycephalum. Microplasmodia that had been labelled with [3H]uridine were used to prepare a surface culture, after a period of growth long enough to ensure that radioactivity was present only in tRNA, rRNA and DNA. The synthesis of rRNA or nuclear DNA during the growth of the surface plasmodium was then followed by measuring the specific activity of the nucleic acid. 2. Synthesis of rRNA during the mitotic cycle shows the following characteristics: (a) it is low during the immediate period of nuclear division, (b) synthesis is then continuous throughout interphase and (c)the rate of synthesis increases 5-6-fold between the beginning and end of interphase. These results are discussed in relation to the pattern of replication of the genes for rRNA. 3. Approximately 80% of the nuclear DNA replicates during the first 90 min of the mitotic cycle; completion of replication, however, occupies the remainder of interphase.


Equilibrium studies on the formaldehyde reaction with native DNA.

Equilibrium analysis of the reaction of formaldehyde with native calf thymus DNA was carried out at temperatures below the thermal transition zone by the spectrophotometric method. The apparent equilibrium constant, Kconf., for the conformational opening and closing reaction of base pairs along the double-helical chain, was measured in various concentrations of formaldehyde, and these values were extrapolated to the zero concentration. The value of KOconf. thus obtained in the absence of the chemical probe was 0.12 in 5 mM 2-(N-morpholino)ethane sulfonic acid buffer (Mes), and 0.003 in 0.15 M NaCl plus 5 mM Mes, pH 7.0, at 50 degrees C. These results make it possible to calculate roughly that 230 base pairs and 7 base pairs are open respectively, at 50 degrees C in the native DNA molecule composed of 2300 base pairs, using Mr = 1.5 x 10(6) for the sample used. This conformational reaction was also characterized by the following thermodynamic parameters: deltaGOconf. = 1.36 kcal - mol-1 (5.68 kJ - mol-1), deltaHOconf. = 36.8 kcal - mol-1 (154 kJ - mol-1), and deltaSOconf. = 110 cal - mol-1 - K-1 (460 J - mol-1 - K-1) in 5 mM Mes, pH 7.0, at 50 degrees C, and the nature of the 'breathing' of base pairs was discussed.