[Preventive measures against fungal infections]. The manifestation of a mycotic infection can be prevented by exposition prophylaxis, disposition prophylaxis or chemoprophylaxis. The main statements are discussed. Protective measurements are helpful provided the instructions are followed continually. One must pay attention to one's personal hygiene. The environmental sanitation can be also important.


[Current knowledge on imperfect and perfect forms of Epidermophyton, Microsporum and Trichophyton species]. The taxonomic status of the members of the form genera: Epidermophyton, Microsporum and Trichophyton is critically appraised. Two species of epidermophyton, 16 of Microsporum and 21 of Trichophyton are considered to be valid. Currently the perfect status of nine Microsporum and eight Trichophyton species have been described. They are respectively classified in the genera Nannizzia and Arthroderma. The value of the tester strains of the heterothallic perfect states in resolving taxonomic problems is discussed.


[The behaviour of middle ear mucosa with cholesteatoma and after tympanoplasty (author's transl)]. The middle ear mucosa of rabbits was studied experimentally. After tympanoplasty inflammatory changes were observed together with an epithelial metaplasia, and the appearance of ciliated and goblet cells was noted. These changes disappeared very slowly over the course of a year when almost normal epithelium was found. Epithelial metaplasia in cholesteatoma was similar to that found following surgery.


[The technique and importance of examination of the function of the maxillary sinus ostium (author's transl)]. A method of measuring the patency of the maxillary sinus ostium under physiological conditions is described. By means of a rhinomanometer passed into the ostium the air-pressure within the sinus with artificially induced pressure changes can be studied. Accurate assessment regarding prognosis and the need to perform sinus surgery can thus be made.


[Problems of hearing aid fitting in elderly adults (author's transl)]. To provide optimum hearing aid fitting in elderly adults, certain factors are necessary which may affect the frequency response curves of the hearing aids. In cases of presbyacusis, an objective fitting of hearing aids can be based on accurate audiometry (via headphone and loudspeaker) with careful interpretation of test results. Importance is placed on directional hearing as it occurs when listening to a speaker against a background of noise. The test methods used in the Department of Audiology, Basel, are described for minitoring the stereophonic fitting of hearing aids. The advantages of this fitting (as opposed to monaural fitting alone) in cases of presbyacusis with symmetrical hearing losses are discussed.


Age and sex difference in response to short exposure to extreme dry heat. Sixty volunteers, 33 males and 27 females (18-63 yr), were divided according to age and sex. They were exposed for 10 min to extreme dry heat: 80-90 degrees C dry bulb temperature and 3-4% relative humidity. Their rectal temperature, skin temperature at eight different points, weight, and heart rate were recorded prior to and immediately following the exposure. A mean rise of only 0.5 degrees C in rectal temperature was recorded following exposure as compared to a mean rise of 5.2 degrees C in mean weighted skin temperature (MWST). Female subjects showed a significantly higher rise in MWST than the male subjects. Similarly, a significantly higher rise in MWST was observed in elderly male subjects as compared to the youngest male group (P less than 0.05). The differences in MWST possibly resulted from differences in mean skin blood flow causing differences in skin conductance. Large individual variation in heat response was recorded in rectal temperature, as well as in weighted skin temperatures. The increase in skin temperature during the first 10 min of exposure to extreme dry heat may serve as an indicator for heat tolerance time, and may help predicting heatstroke susceptible individuals.


Ovarian hormonal responses to exercise.

Ovarian hormonal responses to exercise. The effects of exercise on estradiol, progesterone, follicle-stimulating hormones (FSH), and luteinizing hormone (LH) were studied in nine healthy females. Subjects were studied during light, heavy, and exhaustive exercise in the midfollicular and midluteal portions of their menstrual cycles. Resting hormone levels followed the expected pattern. Increases in estradiol and progesterone occurred at all intensities of exercise in the luteal phase but only in estradiol at exhaustion in the follicular phase. LH was unchanged with exercise in either phase and FSH increased in the follicular phase but not in the luteal phase. We conclude that exercise is a physiological stimulus to elevations in plasma estradiol, progesterone, and FSH, but not LH. The elevations are more marked in the luteal phase for the steroids and in the follicular phase for FSH. Increases in estradiol and progesterone are related to the intensity of exercise and appear to be independent of pituitary control.


Regional distribution of VA/Q in man at rest and with exercise measured with krypton-81m. The regional distribution of ventilation-perfusion ratio (VA/Q) in the lung was obtained from the continuous inhalation and infusion of 81mKr. With this short half-life isotope (13 s) serial measurements were made in six young normal subjects during spontaneous breathing, with minimal radiation exposure. At rest in the upright position the apical zone had a high VA/Q, but over the lower two thirds VA/Q distribution was relatively homogeneous. There was a slight increase towards the base. During exercise at 50 and 100 W the distribution became more uniform but there was more obvious than at rest. These distributions are consistent with a zone of reduced basal blood flow (zone IV) which becomes more marked on exercise. Apical blood flow was followed during the transition from rest to exercise and recovery. There was a large (40-150%) increase within 30 s of starting to pedal at 50 W, but little change thereafter.


Creatine metabolism in skeletal muscle of cold-acclimated rats. Creatine metabolism in skeletal muscle of cold-acclimated rats was studied with the aid of [14C]creatine as a tracer. Creatine contents of all muscles studied were significantly lower in cold-acclimated rats than in controls. Radioactive creatine uptake by soleus of cold-acclimated rats was significantly lower than that of controls, while radioactive creatine uptakes by heart, diaphragm, and gastrocnemius of cold-acclimated rats were not different from those of controls. Urinary creatine dervied from muscle creatine plus urinary creatinine, which corresponds to total creatine release from muscle, increased in the early stage of cold exposure and was restored to control level after completion of cold acclimation. Creatine turnover rate in skeletal muscle was not different between control and cold-acclimated rats. These findings indicate that cold acclimation results in reduced creatine content and uptake of skeletal muscle, and could be interpreted as reflecting enhanced ability of nonshivering thermogenesis in skeletal muscle.


In vivo calibration of flow-dependent blood gas catheters. In vivo calibration of blood gas catheters is complicated by the fact that their signals are flow dependent. A calibration method is described here, which permits continuous in vivo calibration. The principle is to compensate for flow dependence through simultaneous measurement on an inspired, inert reference gas of known partial pressure. The method is demonstrated by experiments on three anesthetized dogs, in which the arterial oxygen partial pressure was measured in situ with a flow-dependent blood gas catheter. The compensated signal was in good agreement with the partial pressure of oxygen measured with conventional techniques.


Wheel running of kangaroo rats, Dipodomys merriami, as related to food deprivation and body composition. Kangaroo rats deprived of food ran themselves to death in 48 h in wheel cages. Despite the loss of 14.5% of body weight the ratio of water to protein was the same after the run as it was in control rats. Metabolic measurements at rest and in the running wheel and weight loss in the 48-h run were used to estimate fuels used and water expended. Two-thirds of the initial amount of fat and 9% of the protein were metabolized. The terminal mean percentage of body fat was about twice that observed in rats trapped in the spring of 1967, when seed production was low: death in the 48-h run could not have been due to depletion of body fat alone. The powerful activity drive seen in hungary kangaroo rats presumably is intensified in dry years when food is scarce and may deplete their reserves enough to result in death from starvation.


Cellular dynamics of rats' submaxillary gland during heat acclimatization. Male laboratory rats (Rattus norvegicus) ranging between 130 and 160 g were subjected to ambient temperatures of 34.5 degrees C from 0 to 28 days. The submaxillary gland, liver, spleen, and kidney were weighed. The acini size and the mitotic index of the acinar cells after intraperitoneal injection of vinblastine sulfate and colchicine were recorded for the submaxillary and parotid glands. The submaxillary glands showed increased growth rate over the initial 5 days, whereas the growth rate of the other organs decreased. The submaxillary gland enlargement was due to an initial hyperplasia of the acini cells reaching a maximum on the 2nd day. This was followed by hypertrophy of the Acini reaching a maximum on the l0th day. The parotid acini at control values throughout the experiment. It is postulated that the gland enlargement is effected to increase evaporative heat loss during the early stages of acclimatization prior to the lowering of heat production resulting from biochemical adaptation.


Canine ventilation after acid-base infusions, exercise, and carotid body denervation. We studied the effect of exercise and carotid body denervation on the ventilatory response which occurs following acute acid-base infusions. Studies were done in 6 dogs prepared with chronic tracheostomies and carotid loops. Ventilation (VE) and arterial pH were measured at rest and during exercise before and after infusions of lactic acid (70 meq), HCl (26 meq), NaHCO3 (45 and 90 meq), or normal saline alone (250 ml). The VE response to [H+] is expressed as 1.min-1/[H+] in nmol.kgH2O-1. Before carotid body denervation (CBD), the response was 0.1 l.min-1[H+] at rest, 1.2 1.min-1/[H+] during exercise. After CBD there was no ventilatory response to [H+] at rest or during exercise. We conclude that 1) Exercise potentiates the [H+] stimulus to breathing. 2) For small changes in arterial [H+], this exercise potentiation is a function of the carotid body. 3) Therefore, that exercise potentiates the carotid body response directly and/or the central projection of this input in the central nervous system. 4) Finally, since carotid body denervation eliminates only 7% (0.8 1.min-1) of ventilation at pH 7.35 in these dogs, that the effective threshold for this ventilatory response approximates a pH slightly greater than 7.35.


Effect of acute and established anemia on O2 transport at rest, submaximal and maximal work. Effects of acute and maintained isovolemic anemia on oxygen transport was studied during rest and exercise in normal males. Following 34% reduction in hemoglobin concentration (Hb), supine and standing Q rose acutely by 56% and 20%, respectively, but returned nearly to the control value by 10-14 days, producing a decrease in PVO2. Redistribution of blood flow appeared to compensate significantly for the decrease in systemic oxygen transport. 2,3-Diphosphoglycerate rose by 18%, in vivo PO2 at half-saturation of hemoglobin (P50) rose by approximately 2 mm over 7-9 days and probably afforded some compensation. The relationship between VO2 and external work intensity was independent of Hb. During exercise, Q/VO2 and VE/VO2 were increased in acute anemia, but PVO2, for a given VO2, decreased below control levels. After 10-14 days the relative increase in VE with exercise persisted; the increase in Q was less pronounced; and PVO2 was further decreased. With both acute and extablished anemia maximal exercise capacity and maximal VO2 (VO2 max) decreased in proportion to the Hb reduction.


Relationship between maximal oxygen uptake and left ventricular function in exercise. Left ventricular systolic time intervals (STI's), cardiac output, and arterial blood pressures were measured during bicycle ergometer work in three groups of 10 young men (aged 20-33 yr) who represented average (VO2max = 42-45 ml.kg-1.min-1), moderate (VO2max = 50-56 ml.kg-1.min-1), and high (VO2max = 59-72 ml.kg-1.min-1) levels of cardiovascular fitness. The subjects were studied using noninvasive procedures at steady-state heart rate of approximately 110, 130, and 150 beats.min-1. At all exercise levels, the fitter subjects displayed slightly shorter values for the preejection period (PEP) compared with less fit groups. Significantly (P less than 0.05) larger stroke volumes, longer left ventricular ejection times (LVET), faster mean systolic ejection rates and lower PEP/LVET ratios were found in the group with the highest aerobic capacity. Although these observations appear to indicate a superior mean level of left ventricular performance in groups who represent high levels of cardiovascular fitness, it was concluded that the use of STI values by themselves to describe the cardiovascular fitness of an individual would not be warranted.


Changes induced by exercise in rates of turnover and oxidation of ketone bodies in fasting man. Seven normal subjects fasted for 3 days were exercised for 30 min on a bicycle ergometer at 60 +/- 30% (mean +/- SE) of their maximal aerobic capacity. Rates of transport and oxidation of ketone bodies were determined at rest and during exercise using a primed constant infusion of [14C]acetoacetate. During the initial 7.5 min of exercise, ketone concentration abruptly decreased (-19.4 +/- 3.1%; P less than 0.001) as the result of a 22.2 +/- 7.0% reduction (P less than 0.05) in ketone production rate (Ra) and a 30.4 +/- 7.5% increase (P less than 0.01) in the rate of uptake (Rd) of ketones. As work progressed, blood ketones continued to fall slowly until the 15th min of exercise and thereafter plateaued at a level which was 21.9 +/- 4.1% lower (P less than 0.005) than resting values. During this apparent new steady state, both Ra and Rd were below control values but tended to be equal and to return simultaneously to their preexercise level. The metabolic clearance rate was increased throughout the entire period of exercise, the rise being more marked during the initial 7.5 min (+40.7 +/- 7.5%; P less than 0.01) than at later periods (+19.0 +/- 7.4%; P less than 0.05). Exercise greatly enhanced ketone oxidation which became virtually complete; despite this, the %CO2 derived from ketones, which averaged 17.6 +/- 1.6% at rest, decreased to 10.1 +/- 0.6% (P less than 0.01) after 30 min of work. This indicates that even during the hyperketonemia of fasting, the increased energy needs related to exercise are met primarily by fuels other than ketones.


Physical training and connective tissues in young mice: biochemistry of long bones. The effect of physical training on collagen, ground substance, and nucleic acid concentrations in long bones was studied in male mice of NMRI strain. The mice to be trained and their controls were about 2 wk old at the beginning of training, which took place on a 5 degrees inclined treadmill 5 days/wk for 3-22 wk. The duration of daily exercise was increased progressively over 3 wk. The final daily exercise bouts were 50 and 80 min for moderate programs and 180 min for the intensive program at a speed of 30 cm/s. Increased concentrations of nitrogen and hexosamines were found at both training intensities, especially after prolonged training. No conclusive changes in nucleic acid concentrations were observed after training. The hexosamine-hydroxyproline ratio was higher and the hydroxyproline-nitrogen ratio lower in the long bones of trained animals compared to the controls. In conclusion these data suggest that prolonged physical activity affects the organic matrix of long bones by maintaining above average concentrations of glycosaminoglycans in matured bones.


Reliability of noninvlasive methods for measuring cardiac function in exercise. Externally recorded systolic time intervals (STI's), indirect (CO2 re-breathing) cardiac output, and auscultatory blood pressures were measured during upright bicycle ergometer work in 20 healthy men, aged 24-56 yr. The subjects were studied on 2 separate days at steady-state work loads chosen to represent light (mean heart rate (HR) = 96 beats.min-1), moderate (HR = 118 beats.min-1) and heavy (HR = 147 beats.min-1) exercise. In addition to determinations of cardiac output, systolic time intervals, and blood pressure, the individual's mean systolic ejection rate (stroke volume/left ventricular ejection time) was calculated as a measure of left ventricular function. In general, heart-rate-corrected STI's were found to be reliable and reproducible. Reliability coefficients for total electromechanical systole and left ventricular ejection time ranged from 0.93 to 0.96 while those for the preejection period and its subcomponents were between 0.63 and 0.88. The reliability of cardiac output, stroke volume, mean systolic ejection rate, and systolic blood pressure was also considered to be acceptable but tended to be higher during moderate and heavy work (r = 0.85-0.95) compared to light exercise (r = 0.60-0.83).


Collateral flow resistance and time constants in dog and horse lungs. We studied collateral flow resistance in exsanguinated, excised lower lobes and accessory lobes of dog and horse lungs, respectively. A double lumen catheter obstructed a peripheral airway isolating a segment of the lobe. Oxygen flowed into the segment via a rotameter which measured flow (Vcoll) while the inner catheter recorded segment pressure (Ps). Gas delivered into the segment flowed out via collateral channels. Collateral flow resistance was calculated as (Ps - PL)/Vcoll, where PL = static transpulmonary pressure. Rcoll at PL = 20, 10, and 5 cm H2O averaged 0.24, 1.25, and 2.65 cmH2O.ml-1.s, respectively, in the dog, and 4.53, 6.00, and 12.62 cmH2O.ml-1.s in the horse. At a given PL, Rcoll measured during inflation. At constant PL, Rcoll increased with time at PL = 5 and 10 cmH2O, but was not time dependent at PL = 20 cmH2O. At constant PL, Rcoll increased at Vcoll increased. We conclude Rcoll is greater in horses than in dogs and is a function of PL, Ps - PL, and lung volume history in both species.


Intravascular volume and tonicity as factors in the regulation of body temperature. The effect of alterations in intravascular volume and tonicity on thermoregulatory and cardiovascular responses to heat and exercise have been compared in four subjects. Core temperatures were found to be significantly higher during dehydration, and when dehydration was prevented by administration of 1% saline, than when dehydration was prevented by water administration. These higher temperatures were associated with elevated levels of plasma [Na] and osmolarity, but no consistent relationship between temperature and changes in intravascular volume could be demonstrated. Relationships observed between core temperature and plasma tonicity were consistent with the hypothesis that the adverse effects of dehydration on thermoregulation can be attributed to an inhibition of sweating mediated by an increase in either plasma osmotic pressure or plasma [Na]. In separate experiments the heart rate response to exercise was shown to be reduced by saline, compared with water and dehydration, and this may be explained by the smaller reduction in intravascular volume which occurs during exercise following administration of hypertonic saline. It is concluded that the effects of reduced intravascular volume, and increased intravascular tonicity on physical work capacity may be distinguished by the adverse effect on the cardiovascular system of the former, and on the thermoregulatory system of the latter.


Potentiation of hypoxic pulmonary vasoconstriction by ethyl alcohol in dogs. Pulmonary and systemic hemodynamics and arterial blood gases were measured in anesthetized and mechanically ventilated dogs before and after oral or intravenous administration of ethanol. Increases in mean pulmonary artery pressure and pulmonary vascular resistance occurred. Platelet antiserum-induced thrombocytopenia inhibition of prostaglandin synthesis with meclofenamate, or alpha-adrenergic blockade did not alter the pulmonary pressor response to ethanol. However, the increase in resistance following ethanol was abolished by hyperoxia and potentiated by hypoxia. Thus, it appears that the effect of ethanol is to augment hypoxic pulmonary vasoconstriction, whereas ethanol per se has no independent pulmonary pressor activity.


Transient O2 uptake response at the onset of exercise. One purpose of this study was to determine whether the half time of the oxygen consumption (VO2) response at the onset of exercise varies with work intensity. Fifteen subjects performed 5 min of bicycle ergometer exercise across a range of relative work loads with VO2 determined continuously via computer analysis. The results revealed that the half time of the VO2 response increased as work intensity increased. Our second purpose was to determine whether VO2 response kinetics are dependent on the maximum oxygen uptake (VO2max) of the subject. When VO2 response kinetics were compared at equal absolute work loads for a highly trained (n = 8, mean VO2max = 70.2 ml/kg.min) and an untrained (n = 7, mean VO2max = 49.7 ml/kg.min) group, the highly trained group had a more rapid VO2 response (P less than 0.05). The same tendency was evident at equal relative work loads but the differences were not significant. The more rapid adaptation of the highly trained subjects at absolute work loads may be caused by the enzymatic and circulatory adaptations which result from endurance training. The more rapid adjustment of VO2 at the onset of exercise may explain the slower phosphagen and glycogen depletion found in highly trained subjects following short-term work at equal absolute loads.


Changes in arterial blood gas tensions during unsteady-state exercise. Arterial oxygen (Pao2) and carbon dioxide (Paco2) tensions and inspired minute ventilation were measured during the first 2 min of stair-climbing exercise in nine normal subjects. The subjects climbed a staircase at a rate of approximately 9 m vertical height every minute and arterial blood was drawn from an indwelling cannula at 15-s intervals. Large falls in Pao2 from a resting value of 92 +/- 2.0 (mean +/- SE) Torr to a lowest value of 65 +/- 3.4 Torr were recorded in the first 50 s of exercise while Paco2 oscillated around the resting value. Most subjects demonstrated an initial plateau of Pao2 for at least 7 s followed by a rapid fall and subsequent rise toward the resting level after 1 min. The falls in Pao2 measured were larger than those reported for laboratory exercise. The possible reasons for this discrepancy are discussed.


Ventilatory and gas exchange responses to cycling with sinusoidally varying pedal rate. To investigate factors controlling ventilation under conditions where the applied work load remains constant, but where hypothesized proprioceptive influences would be expected to vary, five subjects exercised at a constant work rate of 50 W on a cycle ergometer at pedaling rates which varied sinusoidally between 40 and 80 rpm. Each subject exercised continuously for 30 min at each of five sinusoidal periods. Minute ventilation (VE), carbon dioxide output (VCO2), oxygen uptake (VO2), and heart rate were computed breath-by-breath and amplitude and phase relations were extracted. We observed small fluctuations in VCO2 and VO2 engendered by varying metabolic requirements of moving the legs at varying rates. VE fluctuations were closely in phase with VCO2 and the amplitudes of the fluctuations were highly significantly correlated (r = 0.83, P less than 0.001); consequently end-tidal carbon dioxide tension fluctuations were small. Variation of pedaling rate, therefore, did not produce a ventilatory response independent of the effect of VCO2. The ventilatory responses to these forcings are inconsistent with an appreciable role for neurally mediated influences from the exercising limbs and provide further evidence that the exercise hyperpnea is linked to CO2 flow to the central circulation.


Effect of age on survivability of pups eating a carbohydrate-free diet. Seventy-nine pups from 18 litters were changed to a carbohydrate-free diet between 7 and 19 weeks of age. It was found that if the diet change was made after pups were 17 weeks old, the pups remained clinically normal. If the change was made before pups were 17 weeks old, the survival rate was less than or equal to 45.5%.


A comparative study of T-61 and pentobarbital for euthanasia of dogs. A comparative study of T-61 and double-strength pentobarbital for euthanasia of dogs was performed. Effects on EEG, ECG, arterial blood pressure, and respiration were monitored, and time to collapse was measured. In most respects, the effects of the 2 agents were similar; however, 3 of 12 dogs given pentobarbital resumed respiration and cardiac function. None of 9 dogs given T-61 evidenced signs of recovery.


Hemophilia A in three unrelated cats.

Hemophilia A in three unrelated cats. Factor VIII deficiency, consistent with hemophilia A, was detected in 3 unrelated male domestic short-haired cats. Clinical history of each case differed and illustrated the spectrum of clinical manifestations of this disease in cats. Clinical expression of feline hemophilia A was similar to that of man and other animals, in which the degree of factor VIII deficiency tends to determine the severity and frequency of bleeding. All 3 cats had severe protracted bleeding after minor surgical procedures. Hemorrhage was eventually controlled in each case by transfusions of fresh whole blood.


Infectious peritonitis in a cat that subsequently developed a myeloproliferative disorder. The effusive form of feline infectious peritonitis was diagnosed clinically and serologically in a 3-year-old male domestic cat. The cat responded to treatment for 9 months, then developed a myeloproliferative disorder with cytologic characteristics of reticuloendotheliosis.


Intracerebral migration of Cuterebra larva in a kitten. Verminous encephalitis in a 4-week-old kitten was manifested by depression, hysteria, and terminal convulsions. Necropsy revealed a second instar of Cutebra sp in the right cerebral hemisphere. The main lesions in the brain were those of acute focal hemorrhagic encephalomalacia.


The isolation and characterization of narasin, a new polyether antibiotic. Narasin is a new polyether antibiotic produced by a strain of Streptomyces aureofaciens. It is purified by organic solvent extraction and silica gel chromatography. Narasin is active in vitro against gram-positive bacteria, anaerobic bacteria, and fungi and is effective in protecting chickens from coccidial infections.


G2201-C, a new cyclopentenedione antibiotic, isolated from the fermentation broth of Streptomyces cattleya. Streptomyces cattleya produced a new cyclopentenedione antibiotic, G2201-C [C6H6O4(I)], which is moderatley active in vitro against Gram-positive bacteria, weakly active against Gram-negative bacteria, and inactive against fungi. G2201-C is toxic to mice.


A32390A, a new biologically active metabolite. II. Isolation and structure. An inhibitor of dopamine-beta-hydroxylase, designated A32390A, was isolated from the culture broth of a Pyrenochaeta species. The inhibitor showed antimicrobial activity against fungi and gram-positive bacteria. Spectroscopic analysis and chemical degradation studies indicated that the structure was 1,6-di-O-(2-isocyano-3-methylcrotonyl)-D-mannitol.


Quadrone, a new antitumor substance produced by Aspergillus terreus. Production, isolation and properties. A new antitumor compound named quadrone was isolated from the culture broth of Aspergillus terreus NRRL 11,156. Quadrone was active against KB cells in vitro, but died not possess antimicrobial activity.


Chemical modification of some gentamicins and sisomicin at the 3''-position. Chemical and photochemical oxidative methods of de-N-methylation of some gentamicins and sisomicins at the 3''-position are described. Selective acetylation of gentamicins and sisomicins at the 1, 3, 2' and 6' and of gentamicin B at the 1, 3, and 6'positions are achieved by treatment of the free bases with carbon dioxide prior to acetylation. De-N-methylation of the above selectively blocked gentamicins and sisomicins followed by re-alkylation at the 3''-position and de-N-protection gives a series of 3''-N-alkyl analogues. The in vitro antibacterial properties of the new derivatives of gentamicins and sisomicins are given.


The structures of the m-chloroperbenzoic acid oxidation products of 8,9-anhydroerythromycins A- and B-6,9-hemiacetal and of (8S)-8-hydroxyerythromycin B. 13C-NMR studies have confirmed the structures of (8S)-8-hydroxyerythromycins A- and B-6,9;9,11-acetal proposed by KROWICKI and ZAMOJSKI2,3) for the products of the m-chloroperbenzoic acid oxidation of 8,9-anhydroerythromycins A- and B-6,9-hemiacetal. The preparations of (8S)-8-methylthiomethoxy- and (8S)-8-methoxyerythromycin B-6,9;9,11-acetals are described. The latter are stable in aqueous acetic acid under conditions which convert (8S)-8-hydroxyerythromycin B-6,9;9,11-acetal into (8S)-8-hydroxyerythromycin B.


Biotransformation of lankamycin, darcanolide, and 11-acetyllankolide by a blocked mutant of the erythromcyin producing organism Streptomyces erythreus. The biotransformation of lankamycin and congeners darcanolide and 11-acetylankolide by a blocked mutant of the erythromycin-producing organism Streptomyces erythreus, which cannot synthesize erythromycin without supplementation with erythromycin precursors, was investigated. Darcanolide and 11-acetyllankolide were converted into the corresponding 15-deoxy-15-oxo derivatives. Lankamycin was transformed to 15-deoxy-15-oxolankamycin, 4''-deacetyl-15-deoxy-15-oxolankamycin and 3'-de-O-methylankamycin. None of the derivatives possessed high antimicrobial activity.


Sub-unit assembly in the biosynthesis of neomycin. The synthesis of 5-O-beta-D-ribofuranosyl and 4-O-beta-D-ribofuranosyl-2,6-dideoxystreptamines. The preparation of the deoxy- analogues of two pseudodisaccharide fragments of neomycin, 5-O-beta-D-ribofuranosyl-2,6-dideoxy-streptamine and 6-deoxyneamine is described. When added to the growth medium of a deoxystreptamine-idiotroph of Streptomyces rimosus forma paromomycinus only the latter was incorporated into antibiotic, suggesting an obligatory order for the assembly of sub-units. 4-O-beta-D-Ribofuranosyl-2,6-dideoxystreptamine was also prepared. When added to the growth medium of a deoxystreptamine-idiotroph of Streptomyces fradiae it was converted into the 6-deoxyneomycins, apparently after hydrolysis to 2,6-dideoxystreptamine. The structure of the protected derivatives of the ribofuranosyl 2,6-dideoxystreptamines, potentially useful intermediates for the synthesis of novel antibiotics, was shown by using 15C NMR spectroscopy.


Biological and chemotherapeutic studies on three semisynthetic cephamycins. Three semisynthetic cephamycin antibiotics (7alpha-methoxy-cephalosporins), SK&F 73678, SK&F 83088 (CS-1170) and cefoxitin, have been found to possess favorable biological and chemotherapeutic properties. All three cephamycins are active in vitro against strains of Staphylococcus aureus and a variety of gram-negative bacilli. Beta-lactamase producing organisms including indole-producing Proteus spp., Enterobacter spp. and Serratia strains as well as certain anaerobic bacteria were found to be susceptible to these antibiotics. SK&F 73768 showed somewhat better MIC values than cefoxitin against multiple strains of bacteria. Strains of Pseudomonas aeruginosa and group D streptococci are essentially insensitive to these compounds. Their binding to serum proteins is relatively low. In mice, cefoxitin showed the most favorable pharmacokinetics with respect to peak serum level, serum half-life and urinary recovery. These cephamycins protected mice experimentally infected with a variety of bacterial strains. All three compounds are rapidly bacteriolytic to the logarithmically growing Escherichia coli and belatedly so to Staphylococcus strains with complete sterilizing effect. SK&F 73678 and SK&F 83088 showed activity and potency comparable to or better than cefoxitin and thus can be considered candidates for clinical study.


Regulation of the beta-1,3-glucanase system in Penicillium italicum: glucose repression of the various enzymes. The microscopic fungus Penicillium italicum when grown in a synthetic liquid medium produced at least three enzymes with beta-1,3-glucanase activity which were separated by diethylaminoethyl-Sephadex column chromatography. These were named beta-1,3-glucanases I, II, and III respective to their order of elution from the column. A tentative characterization of these three enzymes indicated that they have different modes of action; the first one is an endoglucanase, the second is an exoglucanase, and the third probably has both mechanisms of action. Glucose had a repressive effect on all three enzymes. Only small amounts of beta-1,3-glucanases II and III were present in the cells when they were actively growing in the presence of this sugar. However, when the cells were transferred to a medium low in glucose, a significant increase in the specific activity of beta-1,3-glucanase took place; this was due in part to a much more active production of beta-1,3-glucanases II and III and in part to the appearance of beta-1,3-glucanase I, which could only be detected after more than 12 h of incubation in this medium. The results are discussed in the context of possible beta-1,3-glucanase functions in the fungal cells.


Physiology of lysine permeases in Saccharomycopsis lipolytica. Two active lysine transport systems were detected in Saccharomycopsis lipolytica. No excretion of lysine out of the cells could be obtained, even by chasing with L-lysine or by poisoning with sodium azide. The kinetic properties of one of the permeases, the high-affinity lysine permease, were studied in detail. Its Km was 1.91 +/- 0.23 X 10(-5) M. It proved highly specific, the only potent competitive inhibitors being (i) arginine and its analogs L-canavanine and L-ornithine, and (ii) the lysine analogs L-5 aminoethylcysteine and L-4,5-transdehydrolysine. It is suggested that the high-affinity lysine permease is common to L-lysine, L-ornithine, and L-arginine. The other amino acids tested behaved as noncompetitive inhibitors. The variation of uptake during a growth cycle was studied on ammonia-rich, ammonia-poor, and ammonia-free media. In each case, the uptake exhibited a peak in the early exponential growth phase. No new permease activity was detected during the lag phase or the stationary phase. Ammonia ions competitively inhibited the uptake and also decreased the Vmax value.


Stability of ribosomes of Staphylococcus aureus S6 sublethally heated in different buffers. Cells of Staphylococcus aureus heated at 52 degrees C in magnesium-chelating buffers [pH 7.2, 50 mM potassium phosphate or 50 mM tris(hydroxymethyl)-aminomethane containing 1 mM ethylenediaminetetraacetic acid] leaked 260-nm absorbing material, shown to be RNA, and suffered destruction of their ribosomes. These cells did not regain their salt tolerance when repair was carried out in the presence of actinomycin D (5 microgram/ml). Cells similarly heated in magnesium-conserving buffers [pH 7.2, 50 mM tris(hydroxymethyl)aminomethane containing 10 mM MgCl2 or piperazine buffer] did not leak RNA, suffered no ribosomal damage when heated for 15 min, and recovered, at least partially, in the presence of actinomycin D. Ribosomal damage, is therefore, a consequence of Mg2+ loss and is not an effect of heat per se. Cells suspended in either Mg2+-chelating or Mg2+-conserving buffers lost salt tolerance to about the same extent during heating at 52 degrees C. Therefore, sublethal heat injury can not be attributed to ribosomal damage.


Polyadenylic acid sequences in the RNA of Hyphomicrobium. Heterogeneous RNA containing polyadenylic acid [poly(A)] sequence has been isolated from Hyphomicrobium by affinity chromatography on oligothymidylic acid cellulose and polyuridylic acid Sepharose columns. About 0.1 to 0.3% of [3H]adenine-labeled RNA over a 60-min period is associated with poly(A) sequences. This percentage decreases to about 0.03 in a 20-h labeling period. The poly(A) tracts recovered after digestion with ribonuclease A and T1 are composed of greater than 95% adenine residues and are up to 200 nucleotides in length with a predominant range of 15 to 40 nucleotides. Adenosine and AMP are present in the ratio of 1:36 in alkaline digests of Hyphomicrobium poly(A) tracts. This is compatible with nucleotide lengths determined on acrylamide gels and location at the 3'-OH terminus of the RNA molecule.


Stimulation of lipase production during bacterial growth on alkanes. Acinetobacter lwoffi strain O(16), a facultative psychrophile, can grow on crude oil, hexadecane, octadecane, and most alkanes when tested at 20 but not at 30 degrees C. Growth occurred on a few alkanes at 30 degrees C but after a longer lag than at 20 degrees C. Cells grown on alkanes as sole carbon sources had high levels of cell-bound lipase. In contrast, previous work has shown that those grown on complex medium produced cell-free lipase and those grown on defined medium without alkanes produced little or no lipase. Low concentrations of the detergent Triton X-100 caused the liberation of most of the lipase activity of alkane-grown cells and increased total lipase activity. When ethanol and hexadecane were both present in a mineral medium, diauxic growth occurred; until the ethanol was completely used up, hexadecane was not utilized, and the lipase activity was very low. When growth on hexadecane began, lipase activity increased, reaching a level 50- to 100-fold higher than that of cells growing on ethanol. A similar pattern of lipase formation and hexadecane utilization was observed with Pseudomonas aeruginosa. Whenever A. lwoffi and other bacteria degraded alkanes they exhibited substantial lipase activity. Not all bacteria that produced lipase, however, could attack alkanes. Bacteria that could not produce lipase did not attack alkanes. The results suggest that a correlation may exist between lipase formation and alkane utilization.


Effects of streptomycin and novobiocin on Staphylococcus aureus gene expression. Streptomycin and novobiocin induced production of protein A and inhibited production of alpha- and beta-hemolysins in mutants of Staphylococcus aureus strains RN450 and RN1 resistant to these antibiotics. Streptomycin, but not novobiocin, also inhibited propagation of bacteriophages of serological group B, whereas phages of group A were unaffected. Streptomycin had to be present at adsorption of the phage, and 10 mM CACL2 reversed the inhibitory effect. Lysogenization and competence induction occurred in the presence of streptomycin, suggesting that some early phage genes were expressed.


dnaA acts before dnaC in the initiation of DNA replication. We constructed a double mutant of Escherichia coli K-12 carrying dnaA(Ts) and dnaC(Cs) lesions. In this mutant DNA synthesis proeceeds normally at 32 degrees C and initiation is inhibited at both 41 and 20 degrees C. By shifting this culture grown at 32 degrees C to the two restrictive temperatures in different time sequences and assaying protein and DNA synthesis of cells growing at different temperatures, we found that dnaA and dnaC genes work independently with dnaA acting before dnaC. While preparing special strains for this work, we also showed that the order of genes in the neighborhood of dnaA is dnaA-tnaA-phoS-ilv.


Utilization of oxalacetate by Acinetobacter calcoaceticus: evidence for coupling between malic enzyme and malic dehydrogenase. Growth of Acinetobacter calcoaceticus strain BD413 in malate-mineral medium resulted in the excretion of large quantities of oxalacetate. Malate was virtually depleted by the time the cell density reached 60% of its final value; most of the remaining growth took place at the expense of oxalacetate. Experiments in which oxalacetate was used as the initial substrate showed that pyruvate was not utilized until most of the oxalacetate disappeared. The generation time for growth on malate or oxalacetate was approximately 40 min; the generation time for growth on pyruvate was 62 min, which implies that pyruvate transport may be rate limiting. Oxalacetate and pyruvate, however, supported approximately the same growth yield. These observations suggested that the first step in the utilization of oxalacetate as an energy source consisted of an enzymatic decarboxylation of the keto acid to pyruvate and CO(2). Three enzyme reactions that carry out this decarboxylation have been detected in extracts of A. calcoaceticus. The first, which functioned maximally at pH 4.8, was attributable to the oxalacetate decarboxylase activity of oxidized diphosphopyridine nucleotide-malic enzyme. The second and third, which functioned in the neutral pH range, resulted from coupling of oxidized diphosphopyridine nucleotide-malic enzyme to reduced diphosphopyridine nucleotide-dependent malic dehydrogenase, and oxidized triphosphopyridine nucleotide-malic enzyme to a reduced triphosphopyridine nucleotide-dependent malic dehydrogenase. The efficiency of these coupled reactions was high enough so that the overall reaction could be physiologically significant.


Protein-lipid-lipopolysaccharide association in the superficial layer of Spirillum serpens cell walls. The backing layer of the Spirillum serpens VHA cell wall, which supports and is bonded to the outer, structured protein layer, was isolated and shown to be similar in composition to the same elements of the outer membrane. It contained a lipopolysaccharide that was similar, but not identical, to that of the intact wall and the same phospholipids. The interaction of the isolated wall lipopolysaccharide with the loosely bound wall lipids provided lamellae, whose surfaces were an effective template for a lifelike reassembly of the isolated outer-layer hexagonal protein in the presence of Ca2+. Assembly did not take place on pure lipopolysaccharide, which dispersed in differing forms. A lipid-lipopolysaccharide-water interface appeared to be required as a template surface for the assembly. Lipopolysaccharide from Pseudomonas aeruginosa was able to replace that of S. serpens in the template. These observations suggest that lipid-lipopolysaccharide complexes are highly ordered, and this order is important to the nucleation and assembly of the protein array.


Chemical and structural alterations at the cell surface of Candida tropicalis, induced by hydrocarbon substrate. The surface-localized polysaccharide of alkane-grown cells of Candida tropicalis was identified as mannan containing approximately 4% covalently linked fatty acids. Glucose-grown cells lacked the mannan-fatty acid complex. The surface structure of alkane-grown cells showed a radial arrangement of the wall polymers, with protruding parts. The cell surface of glucose-grown cells was smooth, with a coherent outer limit. The mannan was localized by using concanavalin A. Masking of the mannan with concanavalin A reduced the binding affinity of the surface for alkane, indicating the involvement of the surface-localized mannan-fatty acid complex in the binding of alkanes.


Caulobacter crescentus cell envelope: effect of growth conditions on murein and outer membrane protein composition. The murein and membrane protein compositions of Caulobacter crescentus strains CB13B1a and CB15 have been characterized, and the influence on cell envelope constituents of culture conditions which affect morphogenesis have been studied. Amino acid and sugar analysis of murein sacculi revealed a simple A1gamma murein configuration typical of gram-negative bacteria. The membranes of C. crescentus had low levels of 2-keto-3-deoxyoctonate relative to enteric bacteria, in addition to the absence of lipid A components (Shapiro et al., Science 173:884-892, 1971; Chow and Schmidt, J. Gen. Microbiol, 83:369-373, 1974). Nevertheless, C. crescentus membranes could be fractionated into inner and outer membrane components by sucrose density gradient centrifugation procedures developed for Escherichia coli. The proteins of the outer membrane were distributed between three major (I, II, and III) and two minor (IV and V) protein classes. Class I proteins were greater than or equal to 74,000 daltons and constituted the primary proteins of the outer membrane. Class I proteins were separated into approximately 50 polypeptides by two dimensional gel electrophoresis; the protein composition of thi s class was affected by culture conditions in both CB13B1a and CB15. Class II (47,000 to 39,000 daltons) and III (20,000 to 11,500 daltons) proteins differed in each strain in composition and response to culture conditions.


Identification of an activator protein for myosin light chain kinase as the Ca2+-dependent modulator protein. Myosin light chain kinase which phosphorylates g2 light chain of skeletal muscle myosin requires an activator for the activity (Yazawa, M., and Yagi, K (1977) J. Biochem. (Tokyo) 82, 287-289). This activator has now been identified as the modulator protein known to be a Ca2+-dependent regulator for phosphodiesterase, adenylate cyclase, and ATPases. The identification is based on the quantitative cross-reactivity of muscle activator protein and brain modulator protein in activating myosin light chain kinase and brain phosphodiesterase and identical properties of both proteins in regard to sensitivities to Ca2+, UV absorption spectra, UV absorption difference spectra with or without Ca2+, and mobilities upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In the presence of modulator protein, the activity of myosin light chain kinase was reversibly controlled by the physiological concentration of Ca2+. We suggest that two Ca2+-receptive proteins, i.e. modulator protein and troponin-C, may play roles in the contraction-relaxation cycle of skeletal muscle.


Self-association of glucagon as measured by the optical properties of rhodamine 6G. The fluorescence of rhodamine 6G is completely quenched in glucagon solutions in 0.6 M K2HOP4 at pH 10.6. The absorption of rhodamine 6G is red-shifted by the same reaction. A single rhodamine 6G molecule appears to be bound to a hydrophobic patch in the center of the trimer of glucagon. Since the glucagon monomer has almost no organized structure this site exists only in the associated trimer form of glucagon. The self-association of glucagon to the trimer has been determined from the variation in rhodamine 6G fluorescence and absorption measured over a 60-fold range of dye concentration. The self-association constant agrees with values determined by other methods in the absence of dye. The binding isotherms of rhodamine 6G to glucagon shift with glucagon concentration and exhibit negative cooperativity.


Purification and properties of a thioesterase from lactating rat mammary gland which modifies the product specificity of fatty acid synthetase. An acyl coenzyme A hydrolase (thioesterase II) has been purified to near homogeneity from lactating rat mammary gland. The enzyme is a monomer of molecular weight 33,000 and contains a single active site residue. The enzyme is specific for acyl groups, as acyl-CoA thioesters, containing eight or more carbon atoms and can also hydrolyze oxygen esters. Thioesterase II is capable of shifting the product specificity of rat mammary gland fatty acid synthetase from predominately long chain fatty acids (C14, C16, and C18) to mainly medium chain fatty acids (C8, C10, and C12). Thioesterase II can restore the capacity for fatty acid synthesis to fatty acid synthetase in which the thioesterase component (thioesterase I) has been inactivated with phenylmethanesulfonyl fluoride or removed by trypsinization. No evidence was found of significant levels of thioesterase II in lactating rat liver. The presence of thioesterase II in the lactating mammary gland and the ability of the enzyme to hydrolyze acyl-fatty acid synthetase thioesters of intermediate chain length, are indicative of a major role for this enzyme in the synthesis of the medium chain fatty acids characteristic of milk fat.


Studies on the regulation of leucine catabolism in the liver. Stimulation by pyruvate and dichloroacetate. The rate of oxidation of L-[1-14C]leucine to 14CO2 by isolated rat hepatocytes is increased by pyruvate and dichloroacetate. This effect is specific for L-leucine, not being observed for L-valine, L-isoleucine, or D-leucine. Transamination, the rate-limiting step of L-leucine catabolism in the liver, is the site of stimulation, because uptake of L-leucine by the cells and the oxidation of its transamination product, alpha-ketoisocaproate, are not increased. Measurement of steady state levels of alpha-ketoisocaproate indicate that both pyruvate and dichloroacetate promote the transamination of L-leucine, thereby increasing the availability of substrate for decarboxylation by the alpha-ketoisocaproate dehydrogenase complex (EC 1.2.4.3). Pyruvate stimulation of transamination is secondary to the provision of keto acid acceptors for the amino group of L-leucine. The mechanism of the effect of dichloroacetate remains unknown.


Size and detergent binding of adenylate cyclase from bovine cerebral cortex. Adenylate cyclase from the bovine cerebral cortex has been solubilized with the nonionic detergent Triton X-100. The physical properties of the enzyme have been determined. These are: sedimentation coefficient, 8.1 S; Stokes radius, 70 A; partial specific volume, 0.79 ml/g; mass, 305,000 daltons; f/f0 1.5. The high partial specific volume measured in the presence of detergent indicates that the enzyme binds a large quantity of detergent (0.28mg of Triton X-100/mg of protein). The mass of the enzyme protein alone is 220,000 daltons. The size and detergent-binding properties of adenylate cyclase from bovine cerebral cortex are different from those previously reported for adenylate cyclase from the rat renal medulla (Neer, E. J. (1974) J. Biol. Chem. 249, 6527-6531). The renal enzyme is smaller by 60,000 daltons than the cerebral enzyme and does not bind measurable amounts of detergent. The amount of detergent bound by cerebral adenylate cyclase is characteristic of intrinsic membrane proteins. It suggests that there is a hydrophobic region on the surface of the molecule by which it is held in the plasma membrane.


Rat liver peroxisomes catalyze the beta oxidation of fatty acids. Peroxisomes were purified by differential and equilibrium density centrifugation from the livers of rats treated with clofibrate to enhance their peroxisomal system of fatty acid oxidation. These purified peroxisomes were tested for the presence of crotonase, beta-hydroxybutyryl-CoA dehydrogenase and thiolase using spectroscopic techniques that utilize the characteristic absorption bands of the appropriate 4-carbon acyl-CoA substrates. All three enzymes were found. Analysis of the fractions from equilibrium density centrifugation revealed major peaks of these enzyme activities in peroxisomes and excluded contamination by mitochondria as an explanation of the results. In the presence of excess CoA the purified peroxisomes oxidized palmitoyl-CoA to acetyl-CoA, and reduced NAD, with a 1:5:5 stoichiometry. The peroxisomes were inactive with butyryl-CoA and less active with octanoyl-CoA than with lauroyl-CoA or palmitoyl-CoA; they appear specialized for the beta oxidation of long chain fatty acids.


Metabolic properties of maintained oligodendroglia purified from brain. Oligodendroglia isolated in bulk from dissected white matter of lamb or calf brain can now be maintained in culture for several days. The cells remain intact and viable during this time. The in vitro incorporation of various radiolabeled precursors into lipids or proteins was examined, and its was found that oligodendroglia synthesize the lipids which are predominant in brain, and especially in myelin. They actively synthesize cholesterol, cerebrosides, sulfatides, and all of the phospholipids, especially phosphatidylcholine. In addition, they produce a variety of radiolabeled proteins, with those of high molecular weight exhibiting the most radioactivity. Thus the cells are metabolically very active in vitro and can be used as a model system to study turnover of lipids in the cell, as well as the synthesis of myelin-specific components.


The in vivo stimulation of mannose incorporation into mannosylretinylphosphate, dolichylmannosylphosphate, and specific glycopeptides of rat liver by high doses of retinylpalmitate. Excess vitamin A stimulated the incorporation of mannose into rat liver mannosylretinylphosphate (MRP), dolichylmannosylphosphate (DMP), and into glycoproteins by over 200% during a 20-min labeling period. The glycoproteins were digested with pronase and separated into three components by molecular sieve chromatography. The stimulation of mannose incorporation was greatest in the Peak II glycopeptide (Mr = 6500). In contrast, the incorporation of galactose into glycolipids or glycopeptides was not altered by vitamin A treatment. Analysis of the glycopeptide stimulated by vitamin A treatment showed it to contain mannose, glucose, galactose, and glucosamine in the respective molar ratios of 7:10:17:1 and to be rich in glutamic acid, serine, glycine, aspartic acid, and threonine. The results suggest that excess vitamin A stimulates the incorporation of mannose into glycoproteins by enhancing the synthesis of lipid intermediates involved in specific mannosyl transfer reactions.


Proteinase inhibitors from the excretory gland cells of Stephanurus dentatus. Purification and properties of three secretory proteinase inhibitors. Three proteinase inhibitors designated as I, II, and III were isolated from the excretory gland cells of the swine kidney worm, Stephanurus dentatus. The inhibitors, which were trichloroacetic acid-soluble, were purified by affinity chromatography and ion exchange chromatography. The homogeneity of each inhibitor was shown by polyacrylamide gel electrophoresis and electrofocusing. The molecular weights of the inhibitors estimated by sodium dodecyl sulfate gel electrophoresis fell within a limited range of 9300 to 9700, and the isoelectric points were 6.45, 6.20, and 5.34 for Inhibitors I, II, and III, respectively. The inhibitors formed complexes with trypsin having apparent dissociation constants (Ki) of 2.9 X 10(-11), 7.6 X 10(-11), and 6.4 X 10(-11) M, respectively. Each inhibitor inhibits the esterolytic and proteolytic activities of both trypsin and chymotrypsin. A proteinase inhibitor present in the reproductive organs, intestines, body walls, and esophagi was identical with Inhibitor II found in the excretory gland cells. Culture medium collected after 24-h incubation with adult worms contained the same three inhibitors as the excretory gland cells. These data suggest that the gland cells may secrete the inhibitors internally and externally.


Carnitine biosynthesis. beta-Hydroxylation of trimethyllysine by an alpha-ketoglutarate-dependent mitochondrial dioxygenase. Rat liver mitochondria were found to hydroxylate epsilon-N-trimethyl-L-lysine to produce beta-hydroxy-epsilon-N-trimethyl-L-lysine, an intermediate in carnitine biosynthesis. The hydroxylating system requires alpha-ketoglutarate, Fe2+, and ascorbate, but does not require NADPH nor NADH. No activity was found in the microsomal or soluble fractions of liver extracts. The hydroxylated alpha-amino acid was isolated and characterized by column chromatography using Dowex 50-H+ and Chelex 100-Cu2+ resins and by high voltage paper electrophoresis. The enzymatically produced beta-hydroxy-epsilon-N-trimethyl-L-lysine was shown to be periodate-sensitive and one periodation product was characterized as gamma-butyrobetaine aldehyde. The hydroxylated product was acted upon by crystalline serine transhydroxymethylase (EC 2.1.2.1) to yield gamma-butyrobetaine aldehyde and glycine. Conversion of about 40% of the epsilon-N-trimethyl-L-lysine to beta-hydroxy-epsilon-N-trimethyl-L-lysine was accomplished by this system with little or no further metabolism.


Identification of pyruvate in S-adenosylmethionine decarboxylase from rat liver. S-Adenosylmethionine decarboxylase has been purified to homogeneity (26,000-fold) from rat liver. The enzyme has a molecular weight of 155,000 and a subunit molecular weight of 42,000. One mole of covalently bound pyruvate was found to be present per mole of enzyme subunit. This is the first mammalian enzyme found to contain covalently linked pyruvate.


Influence of 5'-terminal cap structure on the initiation of translation of vaccinia virus mRNA. The ability of methylated vaccinia virus mRNA to bind to ribosomes derived from wheat germ of rabbit reticulocyte lysates has been studied after beta elimination, to remove the 5'-terminal m7G, and after "recapping" of beta-eliminated mRNA molecules using guanylyltransferase.guanine-7-methyltransferase complex from vaccinia virions. Removal of m7G from the mRNA results in significant loss of ability to bind to ribosomes and to simulate protein synthesis in vitro. Readdition of m7G, but not of unmethylated guanosine to the 5' end results in recovery of both of these functions. To evaluate the role of 2'-O-methylation of the penultimate ribonucleoside, mRNAs containing m7G-(5')pppA- and m7G(5')pppG- as well as m7G(5')pppAm- and m7G(5')pppGm- ends were synthesized in vitro at limiting S-adenosylmethionine concentrations by vaccinia virus cores. By comparing the cap sequences of ribosome-bound and unbound mRNAs, we concluded that 2'-O-methylation has at most a minor effect compared to that of m7G upon ribosome binding under in vitro conditions. Only at high input mRNA concentrations, at which competition might occur, was there some ribodomal enrichment of mRNAs containing a specific terminal structure, namely m7G(5')pppAm-.


Cystic angiomatosis (hamartous haemolymphagiomatosis) of bone. A clinicopathological study of three cases. The clinical and radiological features in three cases of cystic angiomatosis of bone are reported. Although these features are generally diagnostic except from histiocytosis X, the definitive diagnosis must be established by a pathological study, preferably of a segment of an involved rib or fibula. The prognosis varies according to the type of clinical presentation-in particular upon whether the lesions are solely skeletal or whether there is extraskeletal visceral involvement. Whereas these last cases may often prove fatal, those with only skeletal involvement have a favourable prognosis: indeed, the cystic bone lesions may regress without any treatment, as occurred in some cases reported in the literature and in two of our three cases.


Points in the technique of recementing in the revision of an implant arthroplasty. When aseptic failure of the bond between the cement and the prosthesis of a total joint replacement occurs, complete removal of the cement has been advocated before the insertion of a new prosthesis. The present study questions this practice. Both laboratory tests on variously prepared specimens of cement and clinical experiences demonstrate that recementing over old cement is a practical alternative if the technique employed includes the removal of blood from the old cement surface, rasping of this surface and the early application of fresh cement.


Partial amputation of the foot for diabetic or arteriosclerotic gangrene. Results and factors of prognostic value. One hundred and eighty-three conservative amputations of some part of the foot in 161 patients with gangrene from diabetes or arteriosclerosis have been studied retrospectively. They constituted 48 per cent of all amputations in one orthopaedic service over a period of twelve years, during which the minimal feasible procedure was always chosen. Sixty per cent healed soundly, but in over a third of these cases at least one revision to a higher level on the foot had been required. Factors that significantly influenced the outcome of the initial operation were the level of amputation, the age of the patient, the interval between the onset of gangrene and operation, anaemia and pyrexia.


Pemberton's acetabuloplasty for congenital dislocation or subluxation of the hip. The results obtained in a consecutive series of thirty-seven Pemberton operations for congenital dislocation or subluxation of the hip are reported. Over the period under review, 1967 to 1973, it was the only type of acetabuloplasty employed at Winford. Unless the mandatory concentric reduction could be obtained with ease, preliminary open reduction was favoured, especially in cases of primary care. Femoral rotation osteotomy was added for marked anteversion. The programme was designed to be complete inside fourteen weeks, and was so for eighteen hips. Secondary acetabuloplasty was performed on hips with instability or dislocation persisting despite previous treatment. The operations were performed from eighteen months to thirteen years of age. One initial failure required a repeat operation which was successful, but one severely dysplastic hip remained so. The average follow-up was six years.


A long-term follow-up of congenital dislocation of the hip. Drawing upon a total experience of 450 hips affected by established congenital dislocation or subluxation, the author presents the long-term results in 177 hips treated for the first time between the ages of nine months and three and a half years, in support of his contention that surgical endeavour should in the first place be directed towards the limbus and upper end of femur rather than the acetabulum. The 144 patients, all treated on lines previously described in this journal (Scott 1953; Somerville 1953a, b; Somerville and Scott 1957), have now been followed up annually for between ten and twenty-five years, both hips receiving equal scrutiny. In brief, the routine has consisted of arthrography, excision of any limbus shown to be inverted, reduction by traction in abduction, and rotation osteotomy of 70 degrees. The addition of 10 to 15 degrees of varus was found beneficial and has become routine. Some hips required secondary procedures, and regret is expressed that these were not carried out sooner. The upper age at which recovery of the acetabulum may occur was found to be much higher than generally supposed, with a critical period between eleven and fourteen. The main conclusion is that in the great majority of cases first seen in this particular age group, improvement of the mechanics of the joint, especially by attention to the upper end of femur, leads to satisfactory development of the acetabulum and good functional results, at least up to early adult life.


The surgical management of kyphosis in older children with myelomeningocele. An operative technique for correction of the severe kyphosis that may develop in cases of myelomeningocele is described. It includes excision of the apical vertebrae, correction of the full length of the kyphotic segment and of rigid compensatory curves, and extensive fusion aided by internal fixation with longitudinal threaded wires. The technique evolved during the management of eleven children. Of these, in the first two the internal fixation employed was inadequate; two others died; and in the remainder good correction was maintained up to the time of review one to six years later.


Proximal femoral focal deficiency.

Proximal femoral focal deficiency. Children with congenital focal deficiency of the proximal femur present many problems that are but rarely encountered by the individual surgeon who is thus unable to accumulate a wide experience. This paper reviews the literature and analyses the treatment of twenty-three cases at the Royal Children's Hospital, Melbourne, with the object of producing a rational plan of treatment. The management of instability of the hip, malrotation, inadequate proximal musculature and leg length inequality are separately considered for five grades of deficiency. Milder forms are amenable to subtrochanteric osteotomy to correct varus deformity. Exploration and grafting of the pseudarthrosis is indicated where progressive deformity develops. In the more severe deficiency, conservative management of the proximal bony defect provides a better result with an operation only rarely indicated. The gross leg length inequality may be most successfully overcome by Syme's amputation with subsequent fusion of the knee to create an above-knee amputee with an end-bearing stump allowing ready fitting of a prosthesis.


Snapping elbow due to dislocation of the medial head of the triceps. A report of two cases. Two cases of unusual snapping at the elbow are described. In both, the medial head of the triceps was found to be separated from the main muscle belly. During flexion of the elbow, the medial head dislocated over the medial epicondyle, producing a characteristic snapping phenomenon. Both cases were of long standing and had been asymptomatic for years. The first clinical symptoms were those of an ulnar neuropathy. In order to restore the normal position of the medial head of the triceps, its tendon was detached from the olecranon, passed under the central tendon and interlaced to it. The ulnar nerve was left in the epicondylar groove in one case and transposed anteriorly in the other. At the end of the procedure flexion of the elbow was unobstructed and the snapping phenomenon had disappeared.


Changes in serum alkaline phosphatase after femoral fractures. Osteomalacia may be a contributory factor in some patients in the development of fractures of the femoral neck and complicate the subsequent management. The level of serum alkaline phosphatase is often valuable in the diagnosis of metabolic bone disease but rises after any uncomplicated fracture, and since such a rise may limit the diagnostic usefulness of this measurement in detecting osteomalacia its extent was assessed in 106 patients. In the majority serum levels were normal on admission, rising after seven to nine days to reach a maximum within a month after fracture. Elevated levels on admission were found in patients with osteomalacia, liver damage or where there had been a delay of several weeks between injury and admission. In a small number of patients normal levels on admission subsequently reached very high values, usually in association with comminution or instability of the fracture. Elevated levels persisted for six to twelve weeks after fracture, the major influence upon the level at this time being the maximum value achieved rather than the presence of osteomalacia. If patients are to be screened for osteomalacia, the alkaline phosphatase must be measured within the first week after a fracture to avoid the distorting influences of the fracture itself.


Patella alta and chondromalacia.

Patella alta and chondromalacia. The Insall-Salvati method of determining the position of the patella by radiography was employed in fifty-one patients with chondromalacia patellae that had been proved at arthrotomy. There was no definite relationship between chondromalacia patellae and patella alta. There was, however, a highly significant statistical difference between the ratio of the length of the patella to that of the patellar tendon in normal men and women.


The surgical treatment of chondromalacia patellae. Four methods of surgical treatment of chondromalacia patellae have been evaluated after periods ranging from two to thirty years (average seven years), to discover the success rate, complications and indications for each. A total of 140 operations had been performed in 98 patients. Overall, satisfactory results were achieved in 25 per cent after forty shavings of the patellar cartilage, 35 per cent after twenty cartilage excisions and drilling of the subchondral bone, 60 per cent after twenty medial transfers of the patellar tendon and 77 per cent after sixty patellectomies. Thirty-four primary patellectomies gave 82 per cent satisfactory results compared with 62 per cent after twenty-six patellectomies performed after a previously unsuccessful operation. The results were worst in patients below twenty years of age especially women and in those with Grade IV changes in the patellar cartilage. Weakness of the quadriceps after any procedure predisposed to an unsatisfactory result. Extensive late radiological degenerative changes in the knee were not seen. On the basis of the results in this report, patellar tendon transfer is recommended in adolescents and athletes with Grade I, II or III changes in the patellar cartilage. In adults over twenty years of age with Grade I and II changes cartilage excision and drilling is satisfactory. In adults with Grade III and adults or adolescents with Grade IV changes patellectomy is the treatment of choice.


The pathogenesis of hallux rigidus.

The pathogenesis of hallux rigidus. The clinical, radiological and pathological features of hallux rigidus affecting nine toes (in seven patients) are described. Characteristic chondral and osteochondral lesions are seen to occur at a specific site on the metatarsal head, and account for the limitation of dorsiflexion but relatively unrestricted plantarflexion typical of hallux rigidus. Radiologically these lesions are often missed because they are mainly cartilaginous and are later obscured by secondary degenerative changes. Histological evidence indicates a traumatic aetiology and a mechanism of injury is suggested.


Total replacement of the first metatarsophalangeal joint. Various prostheses for total replacement of the first metatarsophalangeal joint for painful hallux valgus and hallux rigidus are briefly discussed. Altogether, the results of eighty-six replacements in sixty-nine patients have been recorded after an average interval of two years. In seventy-eight operations a Silastic* prosthesis as designed by Swanson for the replacement of metacarpophalangeal joints was used, with no case of fracture or deep infection up to date. Overall, the assessment of pain showed that 98 per cent of operations gave either complete or considerable relief. For hallux valgus, the objective assessment showed excellent or good results in 79 per cent, fair in 16 per cent and poor in 5 per cent. For hallux rigidus the corresponding figures were 86, 14 and 0. The technique of replacement described promises to be most satisfactory, especially for hallux rigidus. In selected cases of hallux valgus, a basal osteotomy of the first metatarsal should be added.


The problems associated with flail feet in children and their treatment with orthoses. Children with flail feet who have control of their hips and knees are able to walk but are unable to stand still and find it difficult to walk slowly. These problems have been overcome in fifty-three children with flail feet due to myelomeningocele by fitting them with below-knee orthoses which provide maximum stability and yet allow normal walking. The biomechanical principles and the development of the orthoses are discussed.


The management of the foot and ankle in arthrogryposis multiplex congenita. Sixty-six deformities of the foot and ankle in forty-two patients with arthrogryposis multiplex congenita were reviewd. Pes equinovarus proved particularly difficult to treat as there was a tendency for the deformity to recur because of the thick and rigid capsule. Intracapsular procedures were more successful than those done adjacent to the joint. Wide capsulotomy or talectomy were the best procedures under the age of three years and triple arthordesis was most successful for the older child.


Combined application of metabolic myocardial arrest and general profound hypothermia in aortic valve replacement. To determine the value of general hypothermia in combination with magnesium-aspartate-procaine induced metabolic myocardial arrest, the surgical results of 2 similar groups of patients subjected to aortic valve replacement were compared. Metabolic arrest of the myocardium was achieved under mild hypothermic conditions in group I (71 patients) and in profound hypothermia in group II (48 patients). The operative mortality was 5.6% in group I and 4.1% in group II. There was no cardiac related with in group II. In group I two deaths were due to a low cardiac output state. In addition, 3 patients required inotropic support during the early postoperative period. Our results indicate, that magnesium-aspartate-procaine induced cardioplegia in combination with general profound hypothermia can provide effective myocardial protection during aortic valve replacement.


One stage correction of coarctation of the aorta associated with intracardiac defects in infancy. A two stage surgical approach is usually prepared for symptomatic babies affected by coarctation of aorta associated with intracardiac defects, the preliminary operation being the resection of the coarctation. One stage correction has been attempted in six patients, ranging from 2 to 24 m. of age and from 3,2 to 9,7 Kg. of weight. The associated lesions were a large V.S.D. (3 patients), a V.S.D. with pulmonary stenosis, a mitral stenosis and a mitral insufficiency. The coarctation was approached through a left thoracotomy and the intracardiac defect through a midline sternotomy incision, after the patient was turned around. All patients survived and did well following the double procedure. The two-stage surgical approach is unnecessary and probably unadvisable for patients with coarctation of the aorta and associated intracardiac lesions.


Repair of coarctation of the thoracic aorta without resection. Patch graft aortoplasty. Follow-up study of 46 cases. Two hundred and seventy-six patients (276) over the age of one were operated for coarctation of the thoracic aorta from 1949 to 1972. Patch graft aortoplasty, rather than resection of the coarcted segment was the operation of choice in forty-six patients. No hospital or late deaths occurred in this group of patients. Associated defects were present in 20% of these cases. The first patch aortoplasty was performed in 1962. The follow-up study covers a period of 12 years. Haemodynamic study and aortography were performed to assess the long term results. There was no evidence of recoarctation in the follow-up period up to 14 years following surgery and the pressure difference across the repair varied between a minimum of 5 mmHg to 18 mmHg (Table 2). The technique of patch aortoplasty without resection as been extended to a large number of patients and seems to be a very important factor in avoiding a circular constricting anastomosis, preventing the growth of the anastomosis. Maintaining the whole wall of the aorta may also influence the further growth and prevent recurrence in infants and children. Very little surgical dissection is required and suture line tension is also avoided. The technique of patch aortoplasty should be recommended in infants, children and adults with difficult and hazardous dissection.


Pattern formation in early insect embryogenesis - data calling for modification of a recent model. A mathematical model of biological pattern formation based upon lateral inhibition has recently been applied by Meinhardt to insect embryogenesis. This model has stimulated a re-evaluation of previous results, and new experiments designed to test the validity of the model. Split u.v. dose experiments with eggs of the chironomid midge Smittia show that the effective targets for the production of the aberrant 'double abdomen' are not subject to the rapid turnover which is required by the model in its currently published version. Certain types of segment pattern, and differences in the length of segments as predicted by the model could not be observed. Other data conflict with the rather unusual type of photoreversal and the particular view of determination associated with the model. The model can be reconciled with part of the conflicting data if the effective targets for double abdomen induction are regarded as morphogen-producing structures, rather than the morphogen itself which specifies the segment pattern (Meinhardt, personal communication). This version of the model, however, is still at variance with some of the data discussed here. A complementary explanation is proposed taking into account relevant aspects of homoeotic transformations.


Pseudoaneurysm of the axillary artery following a modified Bristow operation: report of a case and review. The Bristow operation with its modifications has become a popular therapeutic modality for recurrent subluxation of the humerus. A patient was recently treated for a false aneurysm of the axillary artery six months after such a procedure. The artery had been lacerated by the dislodged fixation screw. The clinical presentation, intraoperative management and postoperative course were described, and a suggestion for removing the orthopedic screw eight weeks after a Bristow procedure was offered.


The dissociation of nuclear proteins from superhelical DNA. Structures retaining many of the morphological features of nuclei may be released by gently lysing human cells in solutions containing non-ionic detergents and high concentrations of salt. These nucleoids contain superhelical DNA. Using a double-labelling procedure we have compared, at different salt concentrations, the amounts and types of protein associated with human nucleoides containings superhelical or relaxed DNA. We find that the slightly lysine-rich histones (H2A and H2B) but not the arginine-rich histones (H3 and H4) dissociate more slowly from nucleoids containing superhelical DNA than from those containing relaxed DNA. A protein of apparent molecular weight of 22000 also binds more tightly to superhelical DNA. We conclude that this protein and the slightly lysine-rich histones transmute free energy of supercoiling into binding energy when they bind to superhelical DNA.


Echocardiographic evaluation of mitral annuloplasty. Immediate and longterm follow up results. Mitral annuloplasty is preferable to mitral replacement for patients with pure mitral insufficiency. This conservative procedure can be applied to those patients with torn or significantly stretched chordae tendineae unrelated to the site of lesions of the chordae tendineae. The 6 recent recent patients were examined by echocardiography pre- and postoperatively. The study of 35 long term follow up patients revealed longevity and safety of this procedure. Echocardiograms document normalization of abnormal mitral motion, reduction of excursion, and diastolic velocity of the leaflets, and decrease in left atrial size. These findings, showing apparent diminution of mitral flow, suggest significant improvement in valve function.


Involvement of the iliac artery in Buerger's disease (pathogenesis and arterial reconstruction). Out of 216 patients with Buerger's disease, 14 showed the involvement of the iliac artery in addition to the occlusive lesion in the leg arteries. It occurred within 2 to 8 years after the onset of symptom. The obliterative lesion of the iliac artery might owe its cause to a direct proximal progression of thromboangitis obliterans of the femoral artery or to a skip progression of the disease. The skip lesion of the iliac artery might be due to an ascending progress of the thromboangiitic lesion in the branch arteries of the iliac artery. The indication for arterial reconstruction in the case with the involvement of the iliac artery depends on run-off in the deep femoral artery. Out of the 14 cases, 2 underwent thromboendarterectomy with autogenous vein patch grafting of the external iliac artery and 6 underwent bypass grafting, but their long-term follow-up result was poor.


Near fatal hemolysis following repair of ostium primum atrial septal defect. A 15 month old black male who developed near fatal intravascular hemolysis two days after repair of an ostium primum septal defect with a teflon patch is presented. One month after initial operation, reoperation was required for control of hemolysis. At that time a piece of pericardium was placed over the underendothelialized left atrial side of the patch and the mitral cleft was repaired. He initially did well but within several days developed mitral regurgitation. Hemolysis recurred two months postoperatively, but then spontaneously subsided. We propose that the recurrent mitral insufficiency created a defect in the pericardial patch and that caused recurrent hemolysis. Normal endothelial ingrowth probably covered this small defect and hemolysis subsided. Following the second hemolytic episode six months ago, the patient continues to do well.


Assessment of the heart by histochemical and birefringence techniques as compared with ventricular function tests. We have compared results of isovolumic ventricular function studies and results of histochemical and birefringence examinations of biopsy specimens from  80 isolated canine hearts both before and after three hours of preservation utilizing several variables. The direct and derived mechanical tests and histochemical and birefringence assessments of the myocardium were correlated by multiple regression analysis. Our results suggest that initial ventricular health may be indicated by acid hematein staining of free phospholipids and by the response of myocardial birefringence to adenosine triphosphate and calcium, while succinic dehydrogenase staining and birefringence without adenosine-triphosphate treatment are more useful in showing ventricular deterioration during operation.


Effect of intra-arterial reserpine in patients suffering from Raynaud's phenomenon. Several papers have established the beneficial effect of Reserpine applied intra-arterially in patients suffering from Raynaud's phenomenon. In the present study this effect was investigated in 15 patients, some of whom had been submitted previously to sympathectomy, and in healthy controls. A solution of 1.25 mg Reserpine in 10 ml normal saline was injected in the course of one minute in the brachial artery. Blood pressure, pulse rate, bilateral finger venous occlusion plethysmography and fingertip temperature measurements were carried out for two hours following the injection. Later, venous occlusion plethysmographic and clinical controls were continued weekly, until the Reserpine effect had ceased. Normal subjects, patients suffering from Raynaud's phenomenon undergoing no surgical treatment and those submitted to preganglionic sympathectomy showed a conspicuous increase in digital blood flow, while in one patient, who had stellectomy the fingerflow remained unchanged. This effect remained chiefly located on the injected side. There were no blood pressure or pulse rate changes while in the lying position. Hypotension by passive orthostasis was observed two hours after the injection. The possible mechanism of Reserpine is discussed.


The effect of oxygen tension and mechanical stress on the outcome of fresh autogenous tissue implants in the canine blood stream. Fresh autogenous collagenous tissue grafts were implanted in the lumen of the aorta, vena cava, pulmonary artery and vein of thirty dogs. Grafts were studied at varying periods postimplantation. Grafts in the latter three situations were vascularized and absorbed early and were replaced by host fibrous tissue; whereas grafts in the aorta remained unabsorbed because of lack of vascularization. We conclude that fresh autogenous fibrous tissue is not a good material for the reconstruction of the right ventricular outflow tract with a competent valve.


Development of Hydra lacking nerve and interstitial cells. Hydra attenuata were rendered free of interstitial cells (I cells) and interstitial cell derivatives by colchicine treatment. These hydra were then cloned and cultivated for 18 months and their developmental capacities were studied. Some experimental hydra possessed a few (about 1% of the normal numbers) interstitial cells and retained this low level during prolonged culture and active growth without the differentiation of I-cells into specialized cells. Other hydra were completely freed of interstitial cells by the colchicine treatment. Maceration and histological analyses showed that once a hydra is freed of all interstitial cells it does not recover them, nor do its buds contain interstitial cells. I cell-free hydra also lack nerve cells, nematocytes, gametes and endodermal gland cells, and the tissue consists only of ectodermal and endodermal epithelial cells. Hydra completely lacking interstitial cells grow, bud, exhibit tissue renewal patterns, regenerate and preserve polarity generally typical of normal hydra. I cell-free hypostomal tissue has inductive capacity, as does normal hypostomal tissue, when implanted in I cell-free or normal gastric tissue. Regenerating I cell-free tissue undergoes precocious determination as does normal tissue. Only in some quantitative aspects do I cell-free hydra develop abnormally. We conclude that hydra consisting only of epithelial cells are capable of essentially normal development.


A microtubule--nuclear envelope complex in the spermatozoid of Pteridium. The microtubules on the outside of the nucleus of the mature spermatozoid of Pteridium form a complex with the envelope, and the perinuclear space is eliminated. These dissimilar components are held firmly together, possibly by hydrophobic bonding. The outer part of the chromatin is also attached to the inner boundary of the envelope, and pulls away when the envelope is detached.


The cellular origin of chemically induced tumours. Tumours were induced by chemical carcinogens in chimaeric mice made by the amalgamation of two embryos each producing a different electrophoretic variant of the enzyme glucose phosphate isomerase. Although, in these chimaeras, the smallest samples of normal tissue that could be analysed almost invariably contained both isoenzymes, almost all the tumours contained only one. An analysis of the size of the clones that formed the chimaeric epidermis permitted the conclusion that the epidermal tumours produced could not have risen from more than 8 cells; and the most probable interpretation of the data is that these tumours were clonal growths.


The effects of tumour cells on embryonic cell aggregation, adhesion and detachment. The presence of small numbers of tumour cells inhibits the aggregation of embryonic chicken neural retina cells grown in gyratory shaker culture. The aggregation of neural retina cells was also inhibited by ascites cell medium. We investigated whether the inhibitory effect of the tumour cells on aggregate size is effected by inhibition of the initial adhesion or by enhancement of their separation. The number of neural retina cells adherent to microtest plate surfaces was significantly reduced after incubation with either Ehrlich ascites cells or cell-free, conditioned medium, while the percentage of cells removed from glass by shearing was unchanged under those conditions. These results suggest that the reduction in neural retina cell aggregate size produced by Ehrlich ascites cells and their products is due to partial inhibition of neural retina cell adhesion processes, as distinct from enhancement of separation.


Lectin-binding sites are found in rat liver cell plasma membrane only on its extracellular surface. In the present study ultrastructural localization of binding sites for 5 lectins was studied in rat liver cell surface membrane fractions. For this purpose ferritin-coupled Concanavalin A, wheat germ agglutinin, soybean agglutinin, Ricinus communis agglutinin 120 and Lotus tetragonolobus agglutinin I were used as probes for mannose, N-acetyl glucosamine, N-acetyl galactosamine, galactose and fucose moieties in glycoproteins and glycolipids. Although recent reports suggest presence of glycogroups on the cytoplasmic surface of cellular membranes ultrastructural identification of membrane surfaces in the present study indicated an asymmetric localization of lectin-binding sites exclusively on the extracellular side of the membranes.


Genetic analysis of developmental mechanisms in Hydra. II. Isolation and characterization of an interstitial cell-deficient strain. A mutant strain (nf-I) of Hydra magnipapillata was isolated that contained no interstitial cells, nerve cells or nematocytes. This strain appeared spontaneously in a sexually inbred clone of hydra, and it was recognized by its inability to eat. When force-fed, however, it grew, multiplied by budding and regenerated. In this and in many other respects, nf-I was very similar to the interstitial cell-deficient strain produced by Campbell (1976) by means of colchicine. A chimera strain was produced by the reintroduction of interstitial cells from another strain into nf-I. The properties of nf-I, the chimera and other related strains were examined, and the possible roles that the interstitial cells and the nerve cells play in growth and morphogenesis of hydra are discussed.


DNA replication studies in genus Vicia through fibre radioautography. An experiment has been conducted to study the mode of replication in 3 species of Vicia having different C-values by means of fibre radioautography. Characteristics of the units involved in DNA replication of these species have been compared. It has been observed that replication occurs predominantly through bidirectional chain growth, replicating units are arranged in tandem, and replication proceeds from the point of origin in both directions. It has also been shown that there is a positive, though not proportionate, correlation between replication rates, the sizes of replication units and C-values.


Complex nature of serum lysozyme activity: evidence of thermolability in inflammatory bowel disease. In patients with Crohn's disease and ulcerative colitis, alterations in serum storage temperature produced significant changes in serum lysozyme activity (SLA) as measured by the lysoplate method. This was not the case in healthy controls or in a group with other gastrointestinal disorders. Electrophoretic separation of serum revealed two components of lysozyme-type lytic activity but only one in extracts of gut mucosa, leucocytes, and egg white. The major lytic component of serum migrated towards the cathode and reacted with specific antilysozyme serum, but the minor component which migrated towards the anode did not. Although the cause of this anionic lytic activity is uncertain, it contributes to total serum activity as estimated by any method utilising the lysis of Micrococcus lysodeikticus, and may possibly be related to the observed thermolability.


Early detection of folic acid deficiency in elderly patients. Folic acid deficiency is very common in elderly patients who have a mean red cell volume greater than 94 fl. Eighteen out of 40 elderly patients with red cell macrocytosis who initially had normal red cell folate and serum B12 levels subsequently developed folic acid deficiency.


Measurement of red cell folate with 75Se-selenofolate radioassay. 75Se-selenofolate radioassay was adapted for the purpose of red cell folate measurement. The results of assay of 124 whole blood samples by Lactobacillus casei microbiological assay and by radioassay using the adapted selenofolate method were concordant (r = 0.8). Red cell folate levels by the radioassay were below 170 microgram/1 in people who had haematological features of folate depletion. The distribution of results among reputedly healthy subjects extended down to 140 microgram/1.


A whole blood control for the Coulter Model S. The white cell count, red cell count, haemoglobin, and mean cell volume of a new preserved whole blood preparations used as a control material for the Coulter Model S were found to be stable for at least two months. This material provides a cheap and stable whole blood control which is now in routine use in 18 laboratories throughout Wales.


A rapid screening test for reduced fibrinolytic activity of plasma: streptokinase activated lysis time. A simple screening method for determining the fibrinolytic activity of plasma is described. A streptokinase activated system is used, which measures the result of the interplay of all components of the fibrinolytic system with the exception of activators, which are added in excess. Mean lysis time and standard deviation with this method is 6.9 +/- 1.4 min. The sensitivity of the method in detecting minor delays in clot lysis time is demonstrated.


Serum bile acids in patients with hyperlipidaemia. Individual serum bile acids were analysed by an improved gas liquid chromatography method in 12 patients with primary hyperlipidaemia. Total serum bile acid concentrations were raised in 10 subjects. Ursodeoxycholic acid was found in all 12 patients. It was present in significantly greater concentrations, accounted for a greater proportion of the total serum bile acids, and occurred more frequently than in patients with various forms of hepatobiliary disease. Patients with hyperlipidaemia had proportionately less deoxycholic acid than controls but more than patients with liver disease. There was proportionately less chenodeoxycholic acid in patients with hypercholesterolaemia, in whom the primary bile acid ratio was raised.


Idiopathic fibrosis of mediastinum: a discussion of three cases and review of the literature. In two out of three cases of idiopathic fibrosis of the mediastinum there was also retroperitoneal fibrosis, and in one of them extensive fibrosis of the lungs. One of these, a fit middle-aged man, died suddenly with severe coronary artery stenosis due to fibrosis. The third case was that of a young man with a solitary fibrous mass around the right lower lobe bronchus mimicking a malignant tumour and producing bronchial stenosis. Of the various hypotheses about the aetiology and pathogenesis of the fibrous proliferation, that of an autoimmune process seemed the most likely in these cases.


Dysfibrinogenaemia and liver cell growth. From the evidence presented, it is proposed that 'dysfibrinogenaemia' represents the production of normal fetal fibrinogen by rapidly proliferating liver cells in both regenerating and neoplastic tissue. Prolongation of the reptilase clotting time, which was formerly believed to reflect dysfibrinogenaemia, may be rather the result of hepatocytic death.


Morphological and ultrastructural study of the ventral nerve cord in Branchiobdella pentodonta Whitman (Annelida, Oligochaeta). The ventral nerve cord of Branchiobdella pentodonta consists of paired ganglia containing three different types of neurons, a central neuropil showing characteristic synapses and of an intersegmental apparatus made up of two large lateral connectives and a smaller ventral one. Some remarkable differences exist between the ventral nerve cord and that found in other oligochaetes; these include the presence of a large dorsal blood vessel, two dorsal muscular cells per metamere, and supportive glial cells ventral to the neuropil which have long transverse processes coming into contact with the segmental nerves.


Hyperphagia in rats with experimental diabetes mellitus: a response to a decreased supply of utilizable fuels. Alloxan-diabetic rats were hyperphagic when fed diets containing little fat, but they ate normal amounts of food when given diets rich in fat. Normal rats increased food intake to the same degree when the caloric density of their diet was decreased by reducing the content of fats or carbohydrates in isocaloric amounts. Diabetic rats did not respond substantially to changes in caloric density of their diet which were produced by altering the content of dietary carbohydrates, but they systematically increased food intake as the amount of fat in their diet was reduced. Diabetic rats ate normal amounts of a high-fat diet despite continued loss of nutrients in urine and persisting impairments in glucose utilization, fat storage, and liver glycogen deposition. These findings suggest that hyperphagia in experimental diabetes mellitus is a compensatory response to a lack of utiliziable fat fuels rather than the result of a metabolic disturbance per se.


Genetic dominance for low activity in infant mice. Locomotor activity of 2,140 four-day-old inbred, hybrid, and wild mice was measured. Consonant with the prediction that high rates of locomotor activity would be maladaptive in infant mice, hybrids were less active than inbred lines. A triple test cross analysis indicated low heritability and nearly complete dominance toward low activity. Mice from wild stock were even less active than hybrids, which suggests that selection pressures for low infantile activity have relaxed during laboratory domestication. A test is described for estimating changes in selection pressures resulting from laboratory rearing.


Hepatic versus cerebral origin of stimulus for feeding induced by 2-deoxy-d-glucose in rats. Intravenous administration of 2-deoxy-D-glucose (2-DG), a competitive inhibitor of glucose utilization, increased the food intake of rats. Infusions of glucose or mannose abolished this effect, whereas equimolar fructose solutions did not affect 2-DG-induced feeding. Similar results were obtained when 2-DG and the hexoses were administered into the hepatic portal vein. These findings suggest that 2-DG elicits feeding due to glucoprivation at a site that is inaccessible to fructose. This site is likely to be in the brain, not the liver, because all three sugars can nourish peripheral tissue but only fructose cannot penetrate the blood-brain barrier. Moreover, 2-DG-induced feeding was abolished by intravenous infusion of beta-hydroxybutyrate, a substrate that can be oxidized by brain and other tissues but not by the liver.