Laser light-scattering analysis of protoplasmic streaming in the slime mold Physarum polycephalum. Laser light scattering is shown to be an effective means of obtaining a rapid, objective assessment of dynamic changes in the intact plasmodium of the myxomycete Physarum polycephalum during bidirectional (shuttle) streaming. The motion of material in a 100 mum diameter region of a plasmodial vein was studied by following changes in the autocorrelation function of the fluctuations in the scattered light intensity. The autocorrelation function was recorded at 10 s intervals and analyzed to follow changes in the flow velocity of protoplasm associated with shuttle streaming. Rhythmic velocity changes and a "beating" pattern of velocity maxima were readily observed. In an attempt to locate the site of underlying structural changes in the vein responsible for the changing pattern of flow, the average scattered intensity was separated into components derived from moving and stationary scatterers. Periodic variations in the light intensity due to stationary scatterers are related to the streaming cycle and indicate the occurrence of important structural changes in the vein walls. Two possible interpretations of the data are offered; one involving gross dynamic changes in vein structure, the other involving the formation, contraction, or breakdown of fibrillar material in the vein wall during the streaming cycle.


Studies on the in vitro incubation procedure for [35S]sulphate labelling of articular cartilage glycosaminoglycans. Articular cartilage from cow and calf femoral condyles was incubated in Tyrodes solution containing [35S]sulphate for different periods up to 80 min. Glycosaminoglycans from the cartilage tissue and incubation medium were fractionated on Cetylpyridinium chloride and ECTEOLA cellulose microcolumns. The incorporation of [35S]sulphate into all individual fractions of chondroitin sulphate and keratan sulphate was found to be linear from 20 to 80 min incubation time. As a rule the total specific activities of keratan sulphate and chondroitin sulphate were similar for both calves and cows. The proteoglycan material recovered from the medium amounted to about 1% of the tissue dry weight and was found to have a higher chondroitin sulphate: keratan sulphate ratio than the corresponding cartilage tissue for both calf and cow. The solubility profiles for the newly synthesised glycosaminoglycans, obtained from determination of the radioactivity in the individual fractions, were compared with those of glycosaminoglycans already present. These curves indicated that newly synthesised chondroitin sulphate had a higher average molecular size than that present in the tissue whereas the newly synthesised keratan sulphate had a smaller average molecular size. These newly synthesised components were also detected in the proteoglycans recovered from the incubation medium.


Development of Amphioplus abditus (Verrill) (Echinodermata: Ophiuroidea): I. Larval biology. A fraction rich in secretory granules was prepared from the pyloric caeca of Asterias amurensis by sucrose density gradient centrifugation. The freshly prepared fraction exhibited no casein-hydrolyzing activity- but showed nine times as much specific activity as that of tissue homogenates after incubation at 37 degrees C for thirty minutes. Electron microscopy showed that the secretory granules were membrane-bound granules measuring 0.5-290 mu in diameter and contained dense and/or light amorphous substances.


Schizophrenia, brain asymmetry development, and twinning: cellular relationship with etiological and possibly prognostic implications. Discordance for schizophrenia in monozygotic twins has been interpreted as evidence of the role of environment in the etiology fo schizophrenia, with "environment" usually, in practical parlance, carrying Freudian and Sinnerian connotations. MZ twins are often discordant for certain brain development deviations (i) to which they are specially prone, (ii) which show other evidence of having genetic etiologies, and (iii) which involve embryonic symmetry development. MZ twinning is an embryonic symmetry anomaly; its association with others is not surprising. Neurophysiological manifestations of schizoprenia are markedly asymmetric, as is much of normal human mental function. Unusual brain function asymmetry is herein shown to be a factor in virtually every appropriately tested case of MZ discordant schizophrenia.


Acetylcholine-induced current fluctuations in tissue-cultured muscle cells under voltage clamp. Acetylcholine applied ionophoretically to chick skeletal muscle cells grown in tissue culture produces membrane current fluctuations. Cells treated with vinblastine are transformed to a roughly spherical shape. Such transformed cells can be voltage-clamped with microelectrodes. The frequency spectrum of the current fluctuations at fixed voltage obeys a relation of the Lorentz form. From analysis of the current noise, the conductance of a single ionic channel is estimated to be 39 pmho at a temperature of 28 degrees C, and increases with increasing temperature, exhibiting a Q10 of 1.7. The relaxation time for the channel conductance is more sharply temperature dependent, showing a Q10 of approximately 5. These results are in agreement with the picture of acetylcholine-activated ionic channels determined from experiments on frog end plate (Anderson and Stevens, 1973). The relaxation time for carbachol activation is shorter than for acetylcholine, and appears to be more temperature sensitive.


Modification by capsaicin and compound 48/80 of dye leakage induced by irritants in the rat. Concentration-related dye leakage produced by intracutaneous injections of irritants was measured in rats by an Evans blue technique. 2 In rats pretreated with a total dose of 50 mg capasaicin over 4 days, the response to capsaicin, formalin, HCl, KCl, prostaglandin E1, bradykinin and bradykinin with prostglandin E1 (10(-6) M) were greatly reduced, the responses to histamine and 5-hydroxytryptamine were slightly reduced and those to adenosine 5'-triphosphate (ATP) and compound 48/80 were unaffected. 3 Pretreatment with intracutaneous injections of compound 48/80 (0.5 mug, 24 and 48 h previously) recuded the responses to ATP, compound 48/80, HCl, KCl, prostaglandin E1, and bradykinin but did not affect those to histamine, 5-hydroxytryptamine or bradykinin with prostaglandin E1 (10(-6) M). 4 Responses to capsaicin and formalin produced spotted blueing extending over a large area and were suppressed by compound 48/80 in the smaller pretreated area only. Capsaicin responses were reduced with larger doses of compound 48/80 (total dose 15 mug). 5 It is concluded that the production of neurogenic oedema involves both sensory nerves and mast cells.


Inhibitory action of the adenine analog, 4-aminopyrazolo(3,4-d)pyrimidine, in Crithidia fasciculata. The adenine analog 4-aminopyrazolo(3,4-d)pyrimidine inhibits the growth of the kinetoplastid (trypanosomatid) flagellate Crithidia fasciculata. This inhibition is partially overcome only by adenine (of a number of purines tested), with an inhibition index of 0.025. More effective reversal of inhibition is obtained with any of a number of naturally occurring pyrimidine compounds, up to a concentration of 0.18 mM. Higher concentrations of pyrimidines or addition of guanine, as well as adenine and uracil, to the medium increases inhibition. The analog (presumably as the ribonucleotide) was found not to be inhibitory to any enzyme of the pyrimidine biosynthetic pathway that could be tested. It is suggested that the analog competes with adenine for adenine phosphoribosyltransferase (AMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.7), is converted to a ribonucleotide, and is incorporated into nucleic acid.


A xanthine-negative, melibiose-positive species of Streptomyces. The organism VH/CN-71 which had been isolated from a growth in a cat and described initially as an "unusual strain of Nocardia" is a streptomycete. This organism can also produce acid from melibiose although it is unable to decompose xanthine. Its sporophores are short, hooked, curved, or open-looped with extended spirals. Present data suggest that VH/CN-71 may belong to the "flavus" series of Streptomyces.


Establishment of a human leiomyosarcoma cell line. A cell line designated SKN was established from the human uterine leiomyosarcoma of a 52-year-old female. The cell line has grown well and the serial passages were successively carried out 82 times within 12 months. The monolayer cultured cells revealed anaplastic and pleomorphic features, and they multipled rapidly without contact inhibition. Electron microscope studies revealed myoibrils but no virus-like particles, while chromosomal studies showed that all cultured cells were hyperploid, the modal number was 112, and the marker chromosome was present. The cells were transplanted into an immune-depressed hamster cheek pouch and produced a histological leiomyosarcoma resembling the original tumor.


Specialized perinatal care: impact on perinatal mortality. A neonatal intensive care unit was established at one hospital in 1972 when the neonatal mortality was 7.6 and the perinatal mortality 20.9 per 1000 deliveries. In 1973, with full operation of that unit and partial introduction of a high-risk pregnancy unit for fetal monitoring, the rates decreased to 6.4 and 14.9, respectively. With full operation of both units the rates decreased further, to 3.4 and 9.0 in 1974 and 3.8 and 8.9 in 1975. The frequency of cesarean section was 10.1% in 1972-73 and 11.6% in 1974-75. It is concluded that the centralization of obstetric and neonatal care, together with the development of qualified medical and nursing teams, had a major impact in reducing perinatal mortality, and that the frequency of cesarean section was not affected by the introduction of fetal monitoring, although the indications for this precedure became more specific.


Actin-like filaments in the myoid cell of the testis. Microfilaments in the myoid cells of the peritubular tissue in the mouse, swine and human testis bind heavy meromyosin (HMM) and form arrowhead complexes. The periodicity of the arrowhead complexes is about 35 nm. Individual filaments show arrowheads that point in the same direction. Opposing polarity of the HMM-bound filaments is also observed. The microfilaments do not bind HMM in the presence of 10 mM ATP. After treatment with the contraction medium of Hoffmann-Berling, the filaments appear to be undulated. These observations indicate that the microfilaments in the myoid cell are actin-like in nature. A small number of thicker filaments (about 10 nm in diameter) which do not bind HMM is also observed in the cell. Microfibrils which have been reported around the human myoid cell are also found in the swine.


Small angle neutron scattering studies of chromatin subunits in solution. Neutron scattering studies have been performed on dilute solutions of the fundamental subunit of chromatin, the nucleosome. The subunits contain approximately 195 base paris (bp) of DNA and histones H2A, H2B, H3, and H4. Measurements of the small angle scattering curves in various H2O/D2O solvents allow the contrast dependence of the radius of gyration of the subunits to be examined and give the mean scattering density of the particle. Further application of contrast variation to the higher angle scatter curves allows the contributions from the shape and internal structure of the subunits to be analyzed separately. From these results, we are able to propose a spherically averaged structure with most of the histones closely packed into a core of radius 3.2 nm surrounded by a loosely packed DNA-rich shell of 2.0 nm thickness resulting in a particle of 5.2 nm average radius. Model calculations for ellipsoids show that the outer shape of the subunit must have an axial ratio between 0.5 and 1.4 but is probably best described by more spherical particle. These results are correlated with the diffraction from chromatin films to provide an explanation for some of the diffraction rings.


Interference in a radioimmunoassay for human thyrotropin. Abnormally high plasma thyrotropin values were found by radioimmunoassay in some patients when an antiserum to porcine thyrotropin was used, normal results being obtained with an antiserum raised to human thyrotropin. These discrepancies were found in some subjects with no biochemical or clinical evidence of hypothyroidism and occasionally in sera from patients with unequivocal hyperthyroidism. We found a substance in serum that cross reacts with the anti-porcine antiserum, is stable on repeated freezing and thawing, and is independent of the 125I-labeled tracer preparation. It is unlikely that this substance is a separation-stage artefact related to immunoglobulins. Its apparent molecular weight (gel filtration) is 114 000, as compared with apparent molecular weights for standard thyrotropin and endogenous thyrotropin (as found in idiopathic hypothyroidism) of 34 700 and 38 000, respectively. We believe the substance is a normal serum constituent that is present in large quantities in a minority of subjects. Apparently unrelated to TSH, its exact chemical nature remains unidentified.


Autoerthrocyte sensitization---a psychogenic purpura? There are two schools of thought regarding the pathogenesis of this syndrome. Gardner and Diamond and others demonstrated that these patients have a hypersensitivity to extravasated RBCs. They assumed that a fixed tissue antibody reacted with red blood cell stroma producing edema, increased capillary permeability, and further extravasation of blood into the tissues. Ratnoff, Agle, and others, however, believe the syndrome is a psychiatric disorder and that the ecchymotic lesions represent a form of conversion reaction--a mechanism that could explain the patient's myriad complaints. There is evidence that vascular beds are controlled by psychic processes. Neurogenically elaborated kinin-like agents might serve as humoral mediators between the central nervous system and the local tissue reaction.2,4,12 Regardless of the basic etiology, we think the syndrome is more common than has previously been considered and should be thought of in patients with recurrent purpuric eruptions of the extremities.


Acute and chronic elevations in serum levels of FSH after unilateral ovariectomy in the cyclic hamster. During the normal 4-day estrous cycle of the hamster, 12.0 +/- 0.4 (n = 34) ova were shed by the paired ovaries on Day 1. When unilateral ovariectomy (ULO) was performed at 0900 h on any one of the first 3 days of the cycle, approximately the same number of ova were released from the remaining ovary on Day 1 of the ensuing postoperative cycle (compensatory ovulation), except after ULO on Day 4. In view of these results, the principal aim of the present study was to determine if ULO affected acute (24 h postoperative period) and chronic (postoperative cycles) alterations in serum levels of immunoreactive FSH. Multiple bleedings were obtained by cardiac puncture without anesthesia. The acute alteration in FSH levels was first examined at 24-h intervals following ULO at 0930 h on any one day of the cycle, and significant elevations in these levels were found 24 h postoperatively in all but Day-2 hemicastrates. Several experiments were then performed to determine the time-sequence for this 24-h response to ULO. The serum level of FSH was measured at 4-h intervals during the remainder of the cycle in which ULO was done at 0900 h on any one day. The results demonstrated that an acute but transient increase (approximately 100 ng FSH/ml serum) occurred either 7 h or 11 h postoperatively in all the hemicastrates. These transient elevations prior to Day 4 were interpreted to be possible causative factors for the subsequent compensatory ovulation. That they were not the sole cause was evident from the results of another experiment, where compensatory ovulation occurred after ULO at 0030 h on Day 4 (but to a reduced degree; 9.0 +/- 0.6 ova), although serum levels of FSH were not acutely altered. The capability of the remaining ovary to undergo compensatory ovulation after ULO on Day 4 was lost between 0300 h and 0230 h (6.7 +/- 0.8 ova). The serum profile of FSH during the second postoperative cycle after ULO showed a prolonged elevation of FSH levels during the first two days of the cycle compared to the intact estrous cycle. Specifically, the level of FSH gradually declined to 400 ng/ml by 2000 h on Day 1 of the intact (preoperative) cycle but did ont reach this concentration until 2000 h on Day 2 of the postoperative cycle. It is suggested that the prolonged elevated levels of FSH in chronic hemicastrates effect development of a greater number of follicles in the early growth stages in the remaining ovary than the number of follicles per ovary that would otherwise develop in the intact hamster.


A laboratory model demonstrating osteocyte-osteoblast control of plasma calcium concentrations. Table model for plasma calcium control. This report describes a laboratory model designed to illustrate a newer concept for the control of plasma calcium concentrations. This postulate suggests that plasma calcium levels are the result of a balance or imbalance in opposing calcium ion fluxes between plasma and bone fluid compartments existing around osteocyte-lining cells (osteoblasts) units in bone. The metabolic control of these fluxes is postulated to reside within the lining cells on the surface of bone. These cells serve as a cellular interface between the two fluid compartments. The model described in this report illustrates these principles by using the height of columns of water in cylinders to represent calcium ion concentrations. A water pump, representing the calcium transcellular transport system in the lining cells, maintains a higher level of water in the cylinder representing plasma than in that representing bone fluid. This is accomplished by continuous pumping of water from the bone fluid cylinder to the plasma cylinder. Water is returned to the bone fluid cylinder as long as a differential in the height of water in the two cylinders exists. A constant height of water in the plasma cylinder is maintained when the two fluxes are in equilibrium. A constant height of water in the cylinder representing bone fluid is maintained by the level of water in a much larger cylinder representing calcium equilibrium between the solid and liquid phases of bone. The primary hormone controlling the rate of calcium transfer from bone fluid to blood, or in the model regulating the speed of water pump, is parathyroid hormone.


Comparison of the biological effects of thyroxine and triiodothyronine in the rat. To compare the biological effects of thyroxine (T4) and triiodothyronine (T3), the results of varying the production rates of T3 and T4 independently were evaluated. In one set of experiments, the responses of hypothyroid rats to thyroid hormones were measured in terms of weight gain, hepatic mitochondrial alpha-glycerophosphate dehydrogenase (alphaGPD) and serum TSH. T4 was given with, and without, 6-n-propylthiouracil (PTU) and alphaGPD activity paralleled, and could completely be accounted for, by the effect of the quantities of the T3 produced. The direct role of T3 production in the maintenance of hepatic alphaGPD activity was supported by finding normal serum T3 and alphaGPD activities, but reduced T4, in rats on low iodine diet for 2 months. Only after 4 months of iodine deficiency was alphaGPD reduced in the presence of a normal serum T3. These results suggest that T4 per se plays minimal role in this system. In contrast, there were significant effects of T4 administration on stimulation of weight gain and suppression of TSH release in hypothyroid animals which were not due to the T3 produced by peripheral conversion. While T3 given parenterally was about tenfold more potent than T4 in acute suppression of TSH, PTU retreatment did not alter the acute decrease in TSH after T4 which lasted for at least 22 h, as opposed to less than 7 h for T3. Despite the direct effect of T4 on TSH suppression , acute reduction in T3 in normal rats resulted in an elevation of serum TSH even though serum T4 concentrations were unchanged or even increased at this time. The results indicate that the thyrotroph, unlike the hepatocyte, can respond acutely to both increases and decreases in either T3 or T4 production. The differential sensitivities of various tissues to T3 and T4 indicate that the relative potencies of these two hormones must be defined experimentally in terms of a specific biological effect.


The diurnal urinary excretion of oxalate and the effect of pyridoxine and ascorbate on oxalate excretion. The diurnal urinary oxalate excretion has been determined in 11 patients with urolithiasis and in 7 normal subjects. Increased excretion following meals was observed. The variation from hour to hour was most pronounced in the stone patient group. The relation between oxalate concentration and urinary volume was found to follow a biphasic exponential course. Pyridoxine administration increased oxalate excretion in 9 out of 12 subjects and decreased the excretion in 3 subjects. Ascorbate administration increased oxalate excretion in all 7 subjects studied.


Differential activation by some 2-aminotetralin derivatives of the receptor mechanisms in the nucleus accumbens of rats which mediate hyperactivity and stereotyped biting. A series of 2-aminotetralin derivatives were injected into the nucleus accumbens of rat to assess the nature of the dopamine mechanisms in this nucleus which modulate hyperactivity and stereotyped behaviour. It was shown that (1) Derivatives with either 5,6- or 6,7-dihydroxy substitutions were each able to induce hyperactivity and stereotyped behaviour, but substitutions in the 5,6-positions conferred greater potency throughout the series. This differential was emphasised by the continued activity of 2-amino-5,6-dihydroxytetralin in the absence of nialamide whilst the action of 2-amino-6,7-dihydroxytetralin was greatly reduced. (2) The hydroxyl functions in both the 5,6- and 6,7-series were essential for activity: dimethyoxy derivatives were inactive. (3) Generally, substitution of the nitrogen atom with one or two methyl groups, or with a butyl group, reduced or abolished activity. However, N-ethyl and N-propyl substitution markedly enhanced stereotypic potential in the 5,6-dihydroxy series (but not in the 6,7-series). The N-isopropyl derivative in the 5,6-series reflected the activity of the N-propyl compound but a further substitution of the N atom with the methyl group (N-isopropyl-N-methyl) greatly reduced the stereotypic potential without modification of the hyperactivity response. In contrast, N,N-dipropyl substitution abolished the hyperactivity response whilst increasing sterotypic potential. (4) alpha and beta-adrenoceptor blocking agents and alpha-methyl-p-tyrosine failed to reduce the hyperactivity induced by 2-amino-5,6-dihydroxytetralin or the stereotyped behaviour induced by 2-(N,N-dipropyl)-amino-5,6-dihydroxytetralin. Both behaviours were, however, very sensitive to blockade by haloperidol, indicating that both the hyperactivity and stereotyped responses are dopamine-dependent. It is concluded that the dopamine mechanisms in the nucleus accumbens which mediate/regulate hyperactivity and stereotyped behaviour are different. Further, it is suggested that the 2-aminotetralins may be valuable tools in studies designed to assess the topography of cerebral dopamine systems.


Tolerance of haloperidol catalepsy.

Tolerance of haloperidol catalepsy. Haloperidol (0.75 and 1.5 mg/kg p.o.) was administered daily for 16 days to male Wistar rats. The animals received an acute injection of haloperidol (0.5-2.0 mg/kg i.p. or 1.0-4.0 mg/kg p.o.) and catalepsy was measured. After 16 days on haloperidol, all animals became tolerant to the drug, exhibiting decreased cataleptic response to haloperidol; the intensity of catalepsy returned to normal after an additional 16 days abstinence from the drug. In addition, a group of animals treated and tested daily for catalepsy demonstrated that the time course of tolerance development to haloperidol was biphasic, with a rapid phase (T1/2 = 2.5 days) and a slower phase (T1/2 = 5.5 days).


Dietary fiber components: relationship to the rate and extent of ruminal digestion. A mathematical model can serve as a useful reference for describing the mechanisms involved in digestion and for discussing the factors that influence the rate and extent of ruminal digestion. Ruminal digestion can be divided into four components: digestion rate, digestion lag, potential extent of digestion, and passage rate. Each component affects the apparent extent of digestion in a distinct manner and is influenced by separate factors. Digestion rate is directly related to apparent extent of digestion. It is not influenced by chemical entities presently being measured, but may be related to the morphological, crystalline, or physical nature of fiber. It may also be influenced by factors that inhibit or stimulate ruman microbial growth and their fiber-degrading enzymes. Digestion lag is inversely related to apparent extent of digestion; however, factors influencing it are poorly defined. The may include factors affecting microbial populations and their attachment to fiber prior to digestion; or the digestion lag may be related to the chemical or physical alteration of fiber that must occur before digestion can begin. The potential extent of digestion is directly related to apparent extent of digestion and is influenced by plant fiber composition, primarily. Lignin, and possibly silica, functions to limit the potential extent of digestion. Rate of passage essentially competes with rate of digestion for fiber particles as they pass through the rumen; therefore it is inversely related to the apparent extent of digestion. Passage rate is associated with feed intake level and particle size, although other factors such as type of diet and animal physiology may be important.


Glucose transport and recycling determined by means of two tracers and multicompartmental analysis. Glucose labeled with 3H and 14C was injected into cows, and blood samples were taken from 1 to 720 min for measurement of plasma glucose concentration and the specific activities of each tracer. Experiments were done on two cows when nonlactating, lactating, and exhibiting spontaneous ketotic hypoglycemia. Three exponential terms were required to fit the data for each of the tracers in all six experiments. The data were consistent with two 3 compartment models of the glucose system proposed previously on the basis of 14C data only: compartments 1 and 2 represent the glucose pool, while compartment 3 represents derivatives and precurors of glucose. Constraint tests were applied to kinetic parameters in the models to test the hypothesis that [3H]glucose does not recycle. When the parameter representing the recycling of [3H]glucose was set to zero in either model, systematic deviations were found between the predicted and observed data. When the recycling parameters for [3H]glucose and [14C]glucose were allowed to adjust independently of one another, while all other parameters in the model were constrained to be the same for both [3H]glucose and [14C]glucose, a good fit was obtained. These findings are not consistent with the hypothesis that [3H]glucose does not recycle. Recycling of [14C]glucose or [3H]glucose was estimated in terms of the return transport rate back into the glucose pool (compartments 1 or 2) divided by the transport rate from the glucose pool to compartments 3. It was 22 to 53% of [4C]glucose in the normal cows but only 12 and 13% in the ketotic hypoglycemia cows.


Cytogenetic findings in fifty-five couples with recurrent fetal wastage. Balanced chromosomal translocations in parents and Müllerian abnormalities constitute defined causes of reproductive wastage. Fifty-nine couples with histories of recurrent abortion with or without fetal malformations were evaluated with cytogenetic studies and gynecography. In 44 of the couples with pure abortion histories of two or more spontaneous abortions, three (6.8%) balanced carrier parents were identified. In 11 couples with a mixed history of abortion plus fetal malformation, 3 (27.3%) had balanced translocations in one of the parents. The over-all incidence of Müllerian abnormalities in the group of 59 patients was 11.9%.


Twins discordant for 46,XX gonadal dysgenesis. This case report describes twin sisters, one with rudimentary streak gonads and the other with normal ovarian function. Both siblings had normal 46, XX karyotypes, and zygosity testing indicated that they were identical twins. Discordance of identical twins for 46, XX pure gonadal dysgenesis suggests that environmental factors may be a causative factor in some of the karyotypically normal 46,XX forms of gonadal dysgenesis.


Paternal effects on the development of social behavior in Mus musculus. In order to extend the determination and description of effects of the father's phenotype on the development of social behavior in male mice, we tested 24 males from each of 2 highly inbred strains of Mus musculus, A/J and C57BL/6J. Approximately 1 week before the birth of the subjects, we replaced natural fathers with a foster parent such that, within each strain, 8 subjects had preweaning experience with same-strain males, 8 with other-strain males, and 8 with same-strain females, in addition to their natural mothers. Observations of social behavior were conducted at 21, 50, and 60 days of age. The results showed effects of the foster parent's phenotype on the development of social exploration in both strains and, possibly, on agonistic behavior in C57BL/6J mice. The different effects in the 2 strains indicate mechanisms dependent on genotype.


The binding of cations to the surfaces of cells from early chick blastoderms. Cells of the early chick blastoderms are either preparing for or undergoing regulated morphogenetic movements which culminate in the formation of a three-layered embryo. Information on the changes in the physical-chemical properties of cell surfaces may help in the understanding of this process. The binding of magnesium, manganese, strontium, barium and lanthanum to surfaces of early embryonic cells was estimated by the changes induced by these cations in the cells' electrophoretic mobilities (EPM). Cells show a positive EPM at concentrations of MgCl2 and MnCl2 at 3 X 10(-2) M while SrCl2 and BaCl2 were not able to reverse the cells' charge at concentrations up to 6 X 10(-2) M. CaCl3 reversed the cells' EPM at concentrations as low as 5 X 10(-3) M. Our results suggest that the surfaces of early embryonic cells have a high affinity for Mg and Mn. This is indicated by a reversal of polarity which cannot be detected in cells of differentiating or adult tissues at the cation concentrations used in these experiments.


[Microsurgery in gynaecology: endocoagulation, a new method for operations in pelvic endoscopy (author's transl)]. Hemostasis by high frequency current in the abdomen is too dangerous during pelvic endoscopy. Pelviscopy was therefore mainly a diagnostic procedure. Modern electronic coagulation instruments such as the Crocodile forceps and the point coagulator today permit the application of destructive heat without electrical risk for the patient in the peritoneal cavity. The electronic monitor for this coagulation instrument, the endocoagulator gives an optical and acoustical record of the coagulation temperature. The endocoagulation technique permits reorientation of gynaecological surgery to a new micorsurgical technique. Operations for sterilization the lysis of adhesions, the treatment of endometriosis and ovarian cysts in young people reduces the need for laparotomy by 40 to 50% when modern endocoagulation is used. Endocoagulation can now be recommended as routine technqiue for every pelvic endoscopic operation. In 2715 pelvic endoscopies, destructive heat was used in 47% of the cases and the complication rate was 1.3%.


Concept of fibroblast aging in vitro: implications for cell biology. Aging in vitro is defined as the progressive growth decline of mass populations of fibroblastlike cells leading to their eventual loss after serial subcultivations. Different species are compared in terms of the doubling potential of their fibroblasts in culture. A gradient like behavior is found with at one end species whose fibroblasts invariably die after a certain number of doublings and at the other end species yielding fibroblasts with infinite growth potential. The relationship of this phenomenon with cancer and aging is reviewed and discussed. The response of chicken, mouse and human fibroblasts to slow dose rate ionizing radiation fits with the relative positions of these species on the scale relating the probability of yielding fibroblastic populations with infinite life spans. This difference between fibroblastlike cultures is interpreted in terms of the presence or absence of repair mechanisms or of a repairable substrate. The growth potential of fibroblasts in vitro seems to be related to their susceptibility to viral, chemical and physical oncogenes, and could have important implications for the study of evolution.


Effect of nutrient variations on growth and brefeldin A formation in Curvularia lunata. The highest level of secreted brefeldin A was present in glucose-grown cultures, intermediate levels in glucose-fructose, and xylose cultures and low levels in fructose- and galactose-grown cultures of Curvularia lunata. The biomass decreased from glucose, fructose, xylose, glucose-fructose to galactose cultures. Brefeldin A levels and mycelial yields were low in citrate-, gluconate-, and succinate-grown cultures. Inorganic phosphate-limited cultures supported a high level of brefeldin A. Intermediate levels were present in trace elements-, and inorganic phosphate-trace elements-limited cultures.


Conversion of ADP-ribose to 5'-AMP by alkaline treatment and its use for an optical quantitation of mono and poly(ADP-ribose) residues in the micromolar range. ADP-Ribose is nearly quantitatively split to 5'-AMP by treatment with alkali at elevated temperatures. This unique behaviour, which is not shown by ADP and other adenine derivatives, was used as the basis of an optical test for the selective determination of ADPR in the presence of other adenine compounds including RNA. Poly(ADPR) could also be quantified when the polymer was degraded by poly(ADPR) glycohydrolase prior to alkaline treatment. When combined with the determination of the terminal AMP residues released by phosphodiesterase I treatment, the chain length of the polymer could be calculated. Application of the method to the quantitation of protein-bound mono(ADPR) residues in Ehrlich ascites tumor cells under different growth conditions is described.


Medical treatment of the ductus arteriosus. Prostaglandin administered by aortic infusion prevents ductus closure when this is desirable--as in term infants with pulmonary atresia or critical pulmonic stenosis--raising arterial oxygen tension to levels compatible with life. On the other hand, in prematures with persistent patency of the ductus, administering indomethacin inhibits prostaglandin sysnthesis, inducing closure, and can obviate surgery.


Molecular interactions between adriamycin and x-ray damage in mammalian tumor cells. The effect of the anthracycline antibiotic, Adriamycin (Ad), on the sedimentation properties of pre-labelled mammalian DNA has been studied. Ad induces DNA degradation in vivo in both excision repair-competent (HeLa and Me-180) cells and in excision repair-deficient (REQ) cells. When X-irradiated cells are exposed to Ad during the period of repair of DNA single-strand breaks, small numbers of residual breaks persist following completion of repair. These are attributable to those induced by Ad alone. The effects of Ad and X-rays therefore appear to be similar and additive. No clear-cut evidence that Ad can inhibit the repair of X-ray-induced DNA single-strand breakage was found. Ad also induced the formation of DNA double-strand breaks and inhibits the repair of X-ray-induced base damage (repair replication). The induction of DNA strand breakage may be responsible for Ad cell toxicity and may contribute to its capacity to enhance primary X-irradiation damage when the two types of lesions co-exist.


Scanning electron microscopy findings in osteogenesis imperfecta fetalis. A case of osteogenesis imperfecta fetalis in a stillborn black male infant is described. Scanning electron microscopic studies of the skeleton showed normal intercartilaginous septa in the metaphyses of long bones, covered by uniform, calcifying globules 1 mum in diameter. In the diaphyses, however, the orientation of bone trabeculae was abnormal. The trabeculae were thin, with wide and densely packed osteocytic lacunae, and with more resorbing surfaces than in the control specimens. It is concluded that in osteogenesis imperfecta there is abnormal periosteal ossification, together with increased osteocytic resorption.


Pulmonary artery banding for ventricular septal defect in dogs and cats. Five dogs and 1 cat had pulmonary artery banding for ventricular septal defect and congestive heart failure. An umbilical tape band constricted the pulmonary artery to one-third its original diameter, increased the right ventricular pressure, and decreased the left ot right shunt. Five animals had remission of clinical signs; 1 dog died of right ventricular failure due to a band that had been applied too tightly. The results indicated pulmonary artery banding is helpful in reduction of clinical signs, due to increased flow to the lungs from ventricular septal defect.


Streptomyces novoguineensis sp. Nov., an amipurimycin producer, and antimicrobial activity of amipurimycin. A taxonomic study of Streptomyces strain T-36496, which produces an antibiotic effective against rice blast, revealed that it represented a new taxon and it was named Streptomyces novoguineensis sp. nov. The antibiotic, which was named amipurimycin, showed antifungal activity in vitro and considerable curative effect onleaf blast both in green house and field tests at concentrations ranging from 10 to 20 ppm. It was also effective against neck and panicle blast at the same concentration range.


Suppression of amphotericin B methyl ester induced growth stimulation in intraspecific mouse somatic cell hybrids. Amphotericin B methyl ester (AME), the chemically modified derivative of amphotericin B, induced a concentration-dependent growth stimulatory effect on B82 mouse cells as indicated by increased 24- and 72-hour viable cell number, growth rate and DNA and RNA synthesis. In contrast, AME was not growth promoting toward RAG mouse cells or B82-RAG somatic cell hybrids, while hybrid cells exhibited the increased AME resistance pattern of B82 parental cells. A dissociation between the phenotypic expression of growth stimulation and polyene sensitivity was demonstrated in intraspecific mouse hybrids.


Synthesis and biological activity of (2-hydroxy-1-naphthyl)-methylamino acetamido-epicillin and cephradine, and (2-hydroxy-1-naphthyl)-methylacetamido 6-APA. Two new penicillins and a new cephalosporin have been synthesized by condensing 2-hydroxy-1-naphthaldehyde with epicillin, 6-aminopenicillanic acid and cephradine, and subsequently reducing the SCHIFF bases with NaBH4. The antimicrobial activities of these compounds are also described.


Changes in lung mechanics induced by acute isocapnic hypoxia. We measured lung mechanics in seven healthy males during acute isocapnic hypoxia (PAO2 = 40-50 Torr; PACO2 = 38-42 Torr). Hypoxia was accompanied by increases in total lung capacity (mean increase +/- SD; 0.40 +/- 0.24 liters; P less than 0.005) functional residual capacity (0.34 +/- 0.25 liters; P less than 0.01) and residual volume (0.56 +/- 0.44 liters; P less than 0.02) in all subjects. Specific conductance of the lung decreased during hypoxia (P less than 0.02). The static deflation pressure-volume curve of the lung was shifted upward during hypoxia in all subjects. Resting end-expiratory recoil pressure of the lung was slightly, but not significantly lower during hypoxtic expiratory lung compliance was greater during hypoxia (0.39 +/- 0.04 l/cmH2O) than control measurements (0.31 +/- 0.05 l/cmH2O; P less than 0.005). No change was noted in dynamic lung compliance. All changes in lung mechanics were reversed within three minutes of reoxygenation. We conclude that acute isocapnic hypoxia increases total lung capacity in man and suggest that this may be due to the effect of hypoxia on the airways and pulmonary circulation.


Detection of low-level carbadox residues in animal feeds by high pressure liquid chromatography. The high pressure liquid chromatographic (HPLC) method is capable of detecting from 1 to 0.024 ppm methyl 3-(2-quinoxalinyl-methylene)carbazate-N1,N4-dioxide (carbadox). Carbadox is extracted from the feed with 2% NH4OH in acetone, passed through a liquid-liquid partition, subjected to HPLC, and detected by using a 365 nm detector. No feed materials or other active drug ingredients produced false positive results.


A soybean trypsin inhibitor. Crystallization and x-ray crystallographic study. Five trypsin and alpha-chymotrypsin inhibitors which have low molecular weights (ranging from 6800 to 8600) and are present in soybean seeds of the Tracy variety have been isolated and purified, and single crystals which give x-ray diffraction data beyond 3-A spacings have been obtained from one of them. The trypsin inhibitor crystallizes in a monoclinic unit cell of symmetry P2(1) and dimensions a = 25.919(7) A, b = 43.23(1) A, c = 19.905(5) A, and beta = 103.63(2) degrees. The assymmetric unit contains 1 molecule of molecular weight 6800. The crystal, which has been found to be unusually stable to x-radiation, has solvent content of approximately 26% by volume.


Isolation and characterization of a component of the surface sheath of Dictyostelium discoideum. Aggregates of Dictyostelium discoideum are surrounded by a surface sheath which functions to maintain polarity and integrity during development. We have isolated and partially characterized a component of the surface sheath. It is composed of 60% cellulose, 15% protein, 3% heteropolysaccharide (heteropolymer), 5% lipid, and 1% sulfate when isolated from migrating slugs. The sheath, isolated from aggregates prior to tip formation, has less protein, a different heteropolymer, and cellulose of a lower crystallinity than the sheath of migrating slugs. The increase in crystallinity of the cellulose during development may be important in determining the strength of the surface sheath.


Mechanism of action of uridine diphoglucose dehydrogenase. Evidence for an essential lysine residue at the active site. The oxidation of UDP-glucose by the enzyme UDP-glucose dehydrogenase (EC 1.1.1.22) from beef liver has been shown to proceed via the enzyme-bound intermediate, UDP-alpha-D-glyco-hexodialdose. The enzyme does not release this aldehyde, nor can it be trapped by reaction with hydroxylamine, thiosemicarbazide, or cyanide. Tight binding of the intermediate aldehyde can be explained by the recent observation that the essential thiol group of the enzyme forms a thiohemiacetal with the aldehyde during the course of the reaction. However, an enzyme preparation with the essential thiol derivatized with cyanide will still not release the aldehyde, indicating an additional as yet unknown binding mechanism. Derivatization ([14C]formaldehyde, followed by NaBH4 reduction) of 6 of the approximately 168 lysine residues per enzyme molecule (of six catalytic subunits) results in destruction of 47% of the enzyme activity, suggesting the involvement of an essential reactive lysine in the mechanism. Preincubation of the enzyme with UDP-glucose decreases both the loss of activity and incorporation of the label, indicating that this lysine is in the vicinity of the active site. Acid hydrolysis of the labeled preparation, followed by paper chromatography, shows that the label has a mobility, in the system used, that is identical with lysine. Elution of this spot followed by chromatography on Aminex A-5 resin showed that it contained the expected mixture of epsilon-N-methyl lysines. When enzyme that has its essential thiol derivatized with cyanide is incubated with UDP-[14C]glucose and NAD+, and then reduced with NaB3H4, a stable enzyme complex is formed which contains both labels. Acid hydrolysis of this preparation, followed by either two-dimensional paper chromatography or separation in an amino acid analyzer, results in both labels appearing in the position of lysine. It is evident that the enzyme oxidizes the UDP-[14C]glucose to the corresponding aldehyde which occurs as the Schiff's base with an essential lysine. This is then reduced by the NaB3H4 to form a secondary amine which is stable toward hydrolysis and migrates with lysine in separation procedures. As would be predicted, the enzyme can be similarly labeled by treatment with UDP-alpha-D-gluco-hexodisidose alone, followed by NaB3H4 reduction. The same hydrolysis product results from this procedure, and it behaves identically with the product formed by treating alpha-N-acetyl lysine with UDP-alpha-D-gluco-hexodialdose, reducing with NaBH4, and then hydrolyzing. This substance appears to be N5-((5-formyl-2-furanyl)methyl)lysine. When chromatographed on Aminex A-5, both the model compound and enzyme hydrolysate gave peaks corresponding to free lysine and the proposed derivative. Evidence is presented that the oxidation of UDP-glucose to the aldehyde is a concerted reaction involving the formation of the Schiff's base, rather than the formation of the aldehyde with the subsequent formation of the Schiff's base...


Prostaglandin production by methylcholanthrene-transformed mouse BALB/3T3. Requirement for protein synthesis. Stimulation of prostaglandin synthesis in transformed mouse fibroblasts by serum, thrombin, and bradykinin was blocked by actinomycin D and cycloheximide. These RNA and protein synthesis inhibitors did not affect prostaglandin synthetase in vitro or in vivo; nor did they affect the acylation of arachidonic acid into phospholipids. Serum-stimulated release of arachidonic acid and prostaglandins from [3H]arachidonic acid-labeled cells also was inhibited by actinomycin D and cycloheximide. RNA and protein synthesis appear to be required for expression of phospholipase activity; a prerequisite for prostaglandin synthesis by these cells.


Studies on mitochondrial structure and function in Physarum polycephalum. V. Behaviour of mitochondrial nucleoids throughout mitochondrial division cycle. The fine structure of mitochondria and mitochondrial nucleoids in exponentially growing Physarum polycephalum was studied at various periods throughout the mitochondrial division cycle by light and electron microscopy. The mitochondrial nucleoid elongates lingitudinally while the mitochondrion increases in size. When the nucleoid reaches a length of approximately 1.5 mum the mitochondrial membrane invaginates at the center of the mitochondrion and separates the mitochondrial contents. However, the nucleoid does not divide even when the mitochondrial sections are connected by a very narrow bridge. Just before division of the mitochondrion, the nucleoid divides by constriction of the limiting membrane of the dividing mitochondrion. After division, one end of the nucleoid appears to be associated with the inner mitochondrial membrane. The nucleoid then again becomes situated in the center of the mitochondrion before repeating these same processes.


Induction of pinocytosis in rat hepatocytes by partial hepatectomy. Rat hepatocytes, normally not highly pinocytic cells, becomes so after partial hepatectomy when about two-thirds of the liver is removed. Droplets, up to 20 mum in diameter, develop, initially by addition to smaller pinocytic structures and later by fusion with lysosomes. The droplets contain a material with an electron microscope periodicity characteristic of fibrin; they are periodic acid Schiff-positive as is plasma. It is therefore reasonable to consider plasma glycoproteins to be major components of the droplets. The droplets are at all times membrane delimited, an observation possible only after perfusion fixation. The droplets are positive for three lysosomal hydrolases identified cytochemically: acid phosphatase, N-acetyl-beta-glucosaminidase, and beta-glucuronidase. From light and electron microscopy it is evident that these activities are acquired by fusion with lysosomes, mostly autophagic vacuoles and residual bodies both of which become very numerous after partial hepatectomy. Pinocytic structures are seen relatively infrequently in the hepatocytes of normal rats but a great many are present after partial hepatectomy. They are most easily observed if horseradish peroxidase (HRP) is intravenously injected before sacrifice and sections are incubated for HRP cytochemistry. The low dose of HRP employed (10 mg/100 g body weight) does not induce pinocytosis in controls, either untreated rats or rats subjected to laparotomy, including palpation of the liver. However, in partially hepatectomized rats even a much smaller dose of intravenous HRP (3.3 mg/100 g) visualizes the pinocytic structures in hepatocytes (coated vesicles, channels, cuplike bodies, and droplets). Kupffer cells pinocytose much HRP in both control and partially hepatectomized rats.


[A new type of flagellar structure. Type 9+n]. The ultrastructural study of the Eoacanthocephala sperm cell shows a variation from 0 to 5 in the number of the axial fibers in the axoneme. All the species of the order Eoacanthocephala available to us show this variation; moreover, every individual possesses simultaneously several different structural types. So, we are dealing with a new flagellar organization: 9+n, with 0 less than or equal to n less than or equal to 5. In the Quadrigyridae and the Tenuisentidae families, n varies from 0 to 4, with a maximum of 2 for most individuals, exceptionally at 1 for some individuals. In the Neoechinorhynchidae family, n varies from 0 to 5 with a conspicuous prevalence of 3 (from 84 to 99%, according to the individual). These results prompted us to reexamine the two other orders of Acanthocephala in which the structural types 9+2 or 9+0 have been considered as fixed. Indeed, we have found a few flagella the structure of which is different from the prevalent one. It seems, therefore, that the number of the central fibers of the axoneme in the Acanthocephala sperm cell is never absolutely fixed.


Polarity of actin filaments at the initial stage of myofibril assembly in myogenic cells in vitro. The polarity of thin filaments in relation to thick filaments in developing muscle cells in vitro was investigated. The majority of thin filaments exhibited the right polarity and spatial position similar to that seen in mature myofibrils. It appears that the interaction between thick and thin filaments exists in the initial phases of myofibrillogenesis. Cortical microfilaments are found to have their polarities arranged randomly.


Glycoprotein synthesis in a temperature-sensitive Chinese hamster cell cycle mutant. A temperature-sensitive mutant of Chinese hamster cells is described which has two interesting properties: (1) it is a cell cycle mutant and (2) glycoprotein synthesis appears to be affected at the at the non-permissive temerature (40degreesC). Synchronized cells shifed to 40degreesC in the beginning of their G1 phase do not incorporate [3H]-thymidine into DNA during the expected S-phase, but once DNA synthesis has been initiated ( approximately 10 hours after termination of serum starvation) a shift to 40 degrees C no longer leads to an arrest of DNA synthesis. Flow microfluorimetric analysis of DNA content/cell supports this conclusion and indicates that a majority of cells become arrested in the G1 phase of the cell cycle when a non-synchronized population of cells is transferred to 40degreesC. Apparently at all times in the cell cycle there is a drastic reduction if incorporation of labeled sugars (particularly fucose) into glycoproteins. The uptake of fucose and its conversion to GDP-fucose appears to be normal at 40degreesC. Chromatographic analysis indicates that all classes of glycoproteins are affected, and we do not find any evidence for partially completed oligosaccharides at 40 degrees C. Overall protein synthesis is not reduced at he nonpermissive temperature during the time interval under consideration and the number of polysomes attached to membranes (RER) is also normal at 40degreesC. This suggests that the defect is at an early step in the synthesis or regulation of synthesis of glycoproteins. The mutation is a recessive mutation in hybrid cells and mutagen induced revertants can be obtained which grow normally at 40degreesC and in which glycoprotein synthesis at 40 degrees C is restored to normal, wild type levels.


Serum-stimulated phosphate uptake and initiation of fibroblast proliferation. Previous studies have shown that initiation of proliferation of density-inhibited fibroblasts by fresh serum is accompanied by a rapid increase in phosphate uptake. This increase might be a key event in the initiation of DNA synthesis. The present studies examined this possibility. Mouse 3T3, secondary chick embryo, or human diploid foreskin cultures were grown to quiescence in medium containing varying levels of serum. When proliferation of the cultures was initiated by addition of fresh serum, the changes in phosphate uptake were inversely related to the final increases in cell number. Additional experiments showed that the change in phosphate uptake following serum addition was determined by the level of phosphate uptake prior to serum addition. Addition of dexamethasone to quiescent 3T3 cultures caused them to proliferate but did not increase phosphate uptake. Similarly, trypsin or insulin stimulated proliferation of quiescent secondary chick embryo cultures, but caused little or no change in phosphate uptake. Quiescent 3T3 cultures switched to medium containing fresh serum and reduced levels of phosphate showed a decrease in both phosphate uptake and intracellular phosphate pool size. Cell proliferation in these cultures, however, was stimulated to the same degree as cultures switched to medium containing fresh serum and the normal amount of phosphate. In addition, quiescent secondary chick embryo cultures switched to medium containing fresh serum and no phosphate showed a decrease in the intracellular phosphate pool size. Thymidine incorporation and final cell number in these cultures, however, was stimulated to the same or higher degree than in cultures switched to medium containing fresh serum and the normal amount of phosphate. These results demonstrate that the rapid increase in phosphate uptake following addition of fresh serum to quiescent fibroblasts is not a necessary event for the initiation of proliferation.


Production of 5-hydroxyindoleacetic acid from serotonin by cultured endothelial cells. Endothelial cells cells from bovine aorta and human umbilical vein and fibroblasts from human foreskin were cultured and subsequently evaluated for ability to metabolize serotonin (5-HT) to 5-hydroxyindoleacetic acid (5-HIAA). Cells were incubated for three hours with 4 X 10(-6) M [14C] 5-HT creatinine sulfate. [14C] 5-HIAA was separated from labeled 5-HT by column chromatography and measured for scintillation counting. Production of 5-HIAA by bovine aorta cells was 39.0+/-7.5 (S.E.M., n=6) nmoles per 10(9) cells per hour. Production of 5-HIAA was markedly inhibited by the presence of 10(-4) M iproniazid (an inhibitor of monoamine oxidase) or 10(-4) M imipramine (an inhibitor of amine transport). 5-HIAA was the only product of 5-HT metabolism detected by thin layer chromatography. Production of 5-HIAA by human umbilical vein endothelial cells was 5.4+/-2.0 nmoles per 10(9) cells per hour (n=5) and by human foreskin fibroblasts was 3.9+/-1.4 nmoles per 10(9) cells per hour (n=5). The results obtained during incubation in the presence and absence of inhibitors indicate that bovine aorta endothelial cells maintained in tissue culture are able to transport serotonin with subsequent production of 5-HIAA. By contrast, human umbilical vein endothelial cells and fibroblasts exhibited relatively low rates of 5-HT uptake and metabolism.


Differential effects of inhibitors of cell division upon the growth stimulating activities of insulin and serum in nutritionally depleted human and mouse cells. Cultured mouse and human cells were arrested in their growth by artificially depriving them of phosphate. The quiescent cells could be stimulated to synthesize DNA and to divide by addition to the growth medium of insulin, dialyzed serum and/or the full concentration of phosphate. In order to gain insight into mechanisms by which insulin and serum stimulate growth, the inhibitory effects of antimitotic agents were examined. Of the inhibitors tested, vinblastine and cytocalasin B abolished the growth promoting activity of insulin, while colchicine inhibited the activity of both serum and insulin. The present results suggest that insulin-stimulated growth is meciated by a different path way than serum-stimulated growth and is sensitive to mechanisms that occur at various times prior to insulin addition.


The effects of biotin and fatty acids on SV3T3 cell growth in the presence of normal calf serum. The growth of SV3T3 cells in medium containing a low concentration (0.20% v/v) of normal calf serum is enhanced by the addition of biotin or certain unsaturated fatty acids. The biotin effect on the final viable cell density is 5- to 10-fold over the control and is extremely potent, exerting a saturating response at a a concentration of approximately 200 pg/ml. The optimal growth response observed with fatty acids in 5-fold over the control and requires the combination of nervonic acid, palmitoleic acid, and arachidonic acid. The fatty acids are probably not replacing the function of biotin since these two substances are additive in their growth effects.


Secondary activities of diverse inhibitors potentiate the response of hamster embryo cultures to a mitotic stimulus. The reversal of the density dependent inhibition of replication (DDIR) of Syrian hamster embryo cultures by fresh medium containg 30-50% fetal bovine serum was poorly synchronized. There were two waves of DNA synthesis eight to nine hours apart, which by examination of autoradiograms of cultures pulse-labeled with 3H-thymidine, were found to involve 55% and 45% of the cell population. The second wave was found to be due to a subpopulation of epithelioid cells set in the predominantly fibroblastic cell cultures. Pretreatment of the DDIR cultures with hydroxyurea (HU) or arabinosyl cytosine (ara-C), followed by serum stimulation in the absence of the drugs, led to an enhancement of the synchrony. The effect increased with lengthening of the contact with antimetabolites, to a maximum after 20 hours' pre-exposure, and was in part due to the shortening of the GI phase of the epithelioid elements, and in part to increasing the synchrony of the fibroblastic cells. The resulting synchrony involved some 95% of the cells in simultaneous DNA synthesis after a median Gl period of 12 hours. The effect had no relationship to the role of HU and ara-C as specific inhibitors of DNA synthesis since the cultures were mitotically quiescent, and a similar enhanced response could be induced in DDIR cultures by prestimulation exposure lasting only two hours to cycloheximide (cyx), an inhibitor of protein synthesis. Pre-exposure of DDIR cultures to actinomycin D did not potentiate the cell response. A survey of the known secondary inhibitions caused by these three antimetabolites suggests that they all may cause deficiencies in the glycolipid or glycoprotein moieties of the cell surface. These observation provide a use ful, simple means of improving synchrony in these systems and may prove to be a useful probe for investigating the role of the cell surface in regulating cell replication.


Replication kinetics of three DNA sequence families in synchronized mouse cells. Balb/C3T3 cells entered the quiescent Go state following serum deprivation. On addition of fresh serum, more than 95% of the culture resumed growth, but with asynchronous kinetics. If hydroxyurea were added just before the first cells reached S phase, at at least 90% of the cells accumulated at the Gl/S border over the next ten hours. When the block was removed, the culture moved synchronously into S phase. As the cells traversed S, the replication kinetics of three classes of rapidly renaturing DNA were analyzed. Main band highly repeated DNA and foldback DNA replicated continuously, In contrast, satellite DNA replication did not commence until three hours into S, whereupon its rate of synthesis increased ver rapidly, reaching a maximum with the next two hours. These results are discussed in the light of earlier work utilizing other methods of cell synchronization.


Altered morphology and increased cell adhesiveness of chinese hamster ovary cells cultured on fibrin. Chinese hamster ovary cells cultivated on fibrin exhibited different characteristic from cells growing on plastic. While sparsely plated cells on plastic dishes had an epithelioid morphology, cells on fibrin assumed a round shape and then converted to a stretched form with protruded processes that increased with cell density. Within a few days, cells fibrinolysed adjacent fibrin and returned to the morphology seen in plastic dishes. When fibrinolysis was inhibited by epsilon-aminocaproic acid (EACA), cells continued to grow on the fibrin for a longer period and showed dense, criss-crossed fibroblast-type congestion. Whereas, cells on plastic maintained pavement-like epitheloid appearance when they grew to a confluent monolayer. The other altered characteristics on fibrin was increased accumulation of cells in multilayers. Normally as Chinese hamster cells on plastic proliferate, many cells float into the medium instead of piling up after they form a monolayer. On the other hand, cells on fibrin, being maintained by the addition of EACA, remained adherent, piling up multilayers instead of floating into the medium. A possible explanation of these findings is that the surface properties of the stretched cells on fibrin are altered to make them more adhesive. A possible link of these characteristics of the cells on fibrin to tumor cell behavior in vivo is dicussed.


Analysis by high-pressure liquid chromatography of 9-beta-D-arabinofuranosyladenine 5'-triphosphate levels in murine leukemia cells. The metabolism of 9-beta-D-arabinofuranosyladenine (AraA) to arabinofuranosyladenine 5'-triphosphate (AraATP), an inhibitor of DNA synthesis, in mouse leukemia cells was examined by means of high-pressure liquid chromatography. AraATP was separated from naturally occurring nucleotides in acid-soluble extracts and quantitative measurements of AraATP levels were made. A potent inhibitor of adenosine deaminase (2'-deoxycoformycin; co-vidarabine), when used in combination with AraA in the treatment of leukemia-bearing mice, increased the formation of AraATP in mouse leukemia cells four- to five-fold over that obtained by treatment with AraA alone. By means of high-pressure liquid chromatography the half-life of AraATP in tumor cells could be measured. Results of such studies may be of value in planning chemotherapy regimens.


Physiologic control of two neurophysins in humans. Two human neurophysins, nicotine stimulated neurophysin (NSN), and estrogen stimulated neurophysin (ESN) were assayed during physiologic maneuvers and pathologic states in man. NSN is thought to be associated with vasopressin and was elevated in some subjects by volume depletion, surgical stress, hypotension and hypertonic saline infusion. Overnight dehydration did not elevate NSN in spite of urinary concentration. NSN was elevated in some subjects with the syndrome of inappropriate secretion of antidiuretic hormone and when tested was unresponsive to administered water, alcohol or nicotine. ESN was elevated during estrogen administration, in pregnancy, in newborns and in patients with cirrhosis. NSN was also acutely increased at parturition. These data support the association of NSN with vasopressin although changes in NSN were found only with potent stimuli for vasopressin release. ESN may be associated with oxytocin but demonstration of this awaits knowledge of oxytocin physiology in humans.


Effect of long-term calcitonin therapy on the clinical course of osteogenesis imperfecta. 10 children with osteogenesis imperfecta, 4 with "tarda" and 6 with "congenita" varieties of the disease, were treated with salmon calcitonin (SCT) for intervals ranging from 14 to 35 months. Responses to SCT therapy in patients with osteogenesis imperfecta tarda were characterized by an apparent decreased fracture incidence in three, a fall in either alkaline or acid phosphatase, and a rate of increase in forearm bone mass which was greater than that observed in an untreated "tarda" population. The chemical response SCT therapy varied in children with osteogenesis imperfecta congenita, only one demonstrating a decrease in both acid phosphatase and urinary hydroxyproline. Three others responded with a rise in acid phosphatase, two of whom also demonstrated a fall in urinary hydroxyproline; in two other "congenita" patients urinary hydroxyproline was actually higher after SCT treatment and acid phosphatase relatively unchanged. Alkaline phosphatase was normal in all "congenita" patients before and following the SCT treatment interval. These varied biochemical responses were associated with temporary increments in bone mass early in the treatment course, although in bone mass early in the treatment course, although one "congenita" patient with the largest calciuric response to SCT and an increase in hydroxyproline excretion demonstrated progressive increments in skeletal mineral content during a 14-month treatment interval. In both "tarda" and "congenita" subjects, parathyroid hormone was unchanged by chronic SCT treatment; SCT-antibodies were detectable although biological responsivity to SCT persisted.


Influence of dietary protein on susceptibility to alert downer syndrome. Fifty-three dry cows of the Ohio Agricultural Research and Development Center dairy herd were placed in one of four groups for the entire dry period. All received a corn silage-concentrate ration. Groups 1 and 2, however, were maintained on an intake of 8% crude protein while groups 3 and 4 received .65% of their dry matter as calcium and .29% as phosphorus while group 2 received .70% calcium and .70% phosphorus and group 4 received .66% calcium and .65% phosphorus. Animals were bled through the dry period at regular intervals and within 6 h postfreshening or prior to initiation of any therapy. In groups 3 and 4 metabolic disturbance occurred in 69.2% of calvings. These included eight alert downer cows, six of which died during treatment. In groups 1 and 2, the incidence of metabolic disturbance was 7.14% with no downer cows. Blood constituents showed no significant differences except between groups for urea nitrogen of plasma. Individual animals showed no significant changes in blood constituents indicative of a cause of the disorder. Dietary protein influences incidence of the disease. Dietary mineral imbalance and clinical expression of the downer condition were unrelated.


Effects of dominance rank changes, age, and body weight on plasma corticoids of mature dairy cattle. The purpose was to determine whether forced change in dominance rank affected total corticoids of plasma of dairy cattle. Nineteen cows in late lactation were assigned randomly to one of two lots. Restriction of manger space enhanced observations of dominance rank during feeding for three 1-h periods per wk over 6 wk. After 3 wk, a group of the five highest ranking cows from each lot were combined into a new aggressive lot; two groups of subordinate cows formed a docile lot. Blood samples were from tail vein, and concentrations of total corticoids were determined by competitive protein binding. Changes of dominance rank varied from zero to eight places in the 10-cow assemblage after new lots were formed. Body weight was the most significant variable affecting dominance rank. There was no discernable relationship between total corticoids and dominance rank, body weight, or age. The pattern of variation of total corticoids was similar for all groups during the first 3-wk period, but in the second period docile and aggressive groups had dissimilar patterns. Interaction of group by week for total corticoids was significant; however, a cause and effect relationship could not be established. Total corticoids did not appear to be a valid or useful determinant of social stress.


Prepartum serum hormone concentrations related to dystocia in Holstein heifers. Twelve Holstein heifers from a herd having a high incidence of dystocia were bled each day at 0730 h from 260 days gestation until parturition. The blood was analyzed for estradiol-17beta, estrone, 17-hydroxyprogesterone, progesterone, and corticosteroids. Six of the animals had dystocia. The concentrations of estradiol-17beta were lower and the concentration of progesterone higher in dystocial animals during the prepartum period from 23 to 12 days. The prepartum patterns of secretion of corticosteroid, estrone, and 17-hydroxyprogesterone did not change significantly in the two groups between days 23 and 12 prepartum. The prepartum difference in serum estradiol-17beta and progesterone concentration between normal and dystocial animals suggests that the timing of hormone changes in preparation for parturition is delayed and not as pronounced in dystocial animals.


Studies of the mechanism of the human platelet release reaction induced by immunologic stimuli. III. Relationship between the binding of soluble IgG aggregates to the Fc receptor and cell response in the presence and absence of plasma. Aggregated IgG coupled covalently with bis-diazobenzidine (BDB-IgG) and labeled with 3H-diazobenzene (3H-BDB-IgG) has been used to study the binding of soluble IgG aggregates to human platelets in relationship to the release of the contents of intracellular granules (e.g., serotonin). In washed cell suspensions a minimum of 0.14 to 0.2 mug 3H-BDB-IgG per 5 X 10(8) platelets (40 to 70 aggregates per cell) was required for the triggering of the release reaction and cell aggregation. Binding was independent of divalent cations. The Arrhenius plot gave a straight line between 0 to  37 degrees C and a Q10 of 1.6. Neither inhibitors of the release reaction nor energy metabolism, nor formaldehyde fixation of the platelets affected binding. Bound 3H-BDB-IgG was not significantly eluted by IgG, bovine albumin (BSA), buffer, or plasma. Binding to washed platelets was more strongly inhibited by human IgG than by F(ab')2, bovine IgG, human albumin (HSA), or BSA. Plasma was an even more effective inhibitor of both binding and release. Plasma deficient in IgG or depleted of complement retained its inhibitory capacity. In the presence of plasma, at physiologic ratios of plasma and platelets, no release of serotonin was observed. Binding, although inhibited in rate, nevertheless occurred. It was enhanced by divalent cation chelation and had a Q10 of 2.5. The release reaction of washed platelets to which 3H-BDB-IgG had been bound in the presence of HSA or BSA was also inhibited by the subsequent addition of plasma or plasma proteins (human IgG being more effective than bovine IgG,  F(ab')2, HSA, or BSA). 3H-BDB-IgG bound in the presence of either plasma or human IgG did not induce release when the platelets were subsequently suspended in media lacking these proteins. Thus, it appears that the platelet Fc receptor binds 3H-BDB-IgG by a process which is effectively inhibited by plasma, or by free IgG with an intact Fc, and to some extent by high concentrations of other proteins. The effects of bound IgG aggregates are dependent on the other proteins present both during binding and subsequently added. The mechanism of such receptor modulation and its implications in vivo are discussed.


Antibodies against human lymphokines: I. Methods for induction of antibodies capable of neutralizing stable (alpha) and unstable (beta) lymphotoxins released in vitro by activated human lymphocytes. Various methods were employed to induce antibodies in rabbits that were capable of neutralizing different families of lymphotoxins (LT). Both stable (alpha-LT) and unstable (beta-LT) molecules, released by activated human lymphocytes in vitro, were neutralized. The different LT families were first separated into their respective groups by physical-chemical methods. Immunization with small quantities of antigen yielded a high percentage of responder animals. Techniques were developed for eliciting alpha-LT antibodies using as little as 2--3 ml of a cell-free supernatant. The situation was more difficult, however, when the unstable beta-LT molecules were employed as antigens. We found that because of the low concentration and lability of beta-LT in supernatants, the immunizing dose had to be: a) handled rapidly, b) larger than that used with the alpha-LT, and c) injected at closer intervals and over a longer immunization protocol. Physical-chemical studies supperted the concept that the LT-neutralizing activity in the immune serum was immunoglobulin.


Preparation of human antibodies to insulin. Insulin was directly coupled and also indirectly coupled by a side chain to Sepharose 4B to form immunoabsorbents for affinity chromatography of insulin antibodies. The two direct derivatives were insulin lysyl and insulin phenylalanyl sepharose. The indirect derivative, insulin carboxyhexyl sepharose, was prepared in two ways: by a carbodiimide procedure and by an N-hydroxysuccinimide ester method. Insulin carboxyhexyl sepharose prepared by the latter method was found to be the most satisfactory material for affinity chromatography and was used to isolate insulin antibodies from human serum.


Crossed immunoelectrophoresis as applied to studies on complex formation. The binding of heparin to antithrombin III and the antithrombin III--thrombin complex. A two-dimensional immunoelectrophoretic method has been used to obtain information on the binding of heparin to purified antithrombin III and the antithrombin III--thrombin complex. The difference in mobility of the components in a gel containing heparin enables distinction between free and complexed forms of antithrombin III. The results obtained with purified preparations show that heparin is bound more strongly to antithrombin III than to the antithrombin III--thrombin complex. In plasma heparin is bound to several components, only a fraction being bound to antithrombin III. Several components containing antithrombin III are detectable in serum.


Core antigen and circulating anti-core antibody in hepatitis B infection. Core antigen was obtained from the sera of persistent chronic carriers of hepatitis B virus by centrifugation and treatment with Nonidet P40 and 2-mercaptoethanol. The separated core antigen was radiolabelled and identified as a nucleoprotein structure of buoyant density 1.36 g/cm3 and possessing an isoelectric point of 4.4. This material was employed in a radioimmunoassay procedure of high sensitivity for the detection of core antibody. In a series of sera from patients with acute type B hepatitis, core antibody was demonstrated 2 to 3 weeks after the onset of jaundice during the period of surface antigenaemia. The presence of core antibody may therefore provide an accurate serological marker for the detection of active or recent virus replication in future epidemiological studies of hepatitis B infection.


The structure of infectious pancreatic necrosis virus RNA. The RNA from infectious pancreatic necrosis virus has been purified and had a sedimentation velocity of 14S on sucrose gradients, a buoyant density of 1-60 g/ml in CS2SO4 and pyrimidine to purine ratios near unity. The RNA had the appearance of a linear double stranded molecule with an average length of 0-92 mum and a standard deviation of 0-07 mum when observed under the electron microscope using the Kleinschmidt protein film technique. This would correspond to a mol. wt. of 2-4 +/- 0-2 X 10(6). The RNase A resistance of IPN virus RNA exhibited a marked salt dependence; it was 92% resistant in 0-1 M-NaCl, but only 9% resistant, or less, in 0-*1 M-NaCl. The RNA was resistant to denaturation by boilding at NaCl concentrations of 0-04 M or higher, but did denature at lower concentrations. Polyacrylamide gel electrophoresis of the RNA indicated that two RNA species were present and the standard deviation of lengths in the electron microscope indicated that they could not differ by more than 4 X 10(5) in mol. wt.


Polykaryocytosis induced by certain arboviruses in monolayers of BHK-21-528 cells. Multinucleated giant cell formation in a clone of BHK-21 cells, BHK-21-528, was tested with certain arboviruses. Eleven out of 19 viruses tested, Chikungunya, Getah, Sagiyama, Sindbis, Western equine encephalitis, St. Louis encephalitis, Japanese encephalitis, West Nile, Bunyamwera, Germiston and California encephalitis virus induced cell fusion after infection. All cases where giant cells were observed during the experiments, involved the B type cell fusion (fusion from within).


Acquired drive strength as a joint function of UCS intensity and number of CS-UCS pairings. The form of the empirical law relating acquired drive strength to UCS intensity and number of CS-UCS pairings was investigated in two experiments using 120 and 30 male albino rats, respectively. Strength of UCS was varied at a broad range of intensities. Controls were introduced to prevent the intrusion of competing crouching responses (R1) on the measurement of response strength (R2). Acquired drive strength was found to be curvilinearly related to UCS intensity and number of CS-UCS pairings during conditioning. The relationship was monotonic and negatively accelerated. Habit strength, or sHr, was found to be jointly determined by UCS intensity (D) and number of CS-UCS pairings during aversive conditioning. These results were contrasted with findings for the instrumental appetitive learning case of Hull's system, where it has been shown that sHr and D are independent and interact in a multiplicative fashion in their effect on performance. An historical review of acquired drive experiments was also presented.


Electron microscopic studies on embryonic chick spinal ganglion cells: relationship between microtubules and the Golgi complex. Spinal ganglia from 11 day chick embryos were fixed immediately after removal, or after a short incubation in media with or without nerve growth factor (NGF), and were subsequently examined by transmission electron microscopy. Qualitatively, incubation did not effect the fine structure of the neuroblasts. Morphometrically, however, NGF was found to cause a marked increase in the amounts of microtubules and microfilaments in the perikarya of the cells with a threefold increase in the volume density of these organelles after 4 h. The cells displayed a prominent Golgi complex, mostly with dictyosomes organized in one continuous band or area. Individual dictyosomes consisted of a stack of 3-6 parallel cisternae associated with small vesicles and occasional larger vacuoles. Microtubules were observed in all parts of the cyotplasm but were particularly numerous within the Golgi area. They occurred both on the forming and maturing sides of the dictyosomes and in the latter site were closely associated with small vesicles and peripheral dilatations of the cisternae. These observations indicate a possible role of microtubules in the organization and function of the Golgi complex.


Electron microscopic studies on embryonic chick spinal ganglion cells: in vitro effects of antimicrotubular agents on the Golgi complex. Spinal ganglia from 11 day chick embryos were incubated in media containing colchicine or vinblastine and subsequently examined by transmission electron microscopy. Both drugs caused a partial disappearance of cytoplasmic microtubules and a concomitant growth in the number of microfilaments in the neuroblasts. In ganglia treated with vinblastine for 4 h these effects were recorded morphometrically as a decrease of about 80% in the volume density of microtubules and a more than tenfold increase in the volume density of microfilaments. Furthermore, the cells displayed marked structural changes in the Golgi complex. The dictyosomes were mostly distinctly separated from each other and individual dictyosomes showed a decrease number of cisternae and an increased number of closely associated large vacuoles. The results are discussed with regard to the role of microtubules in the organization and function of the Golgi complex in nerve cells.


Immunoproteins in human brain tumor cyst fluids. Quantitation of the concentration of immunoproteins in serum, and cystic fluids from six patients (three with cerebral astrocytoma and three with cerebellar hemangioblastoma) has been determined. The values for total protein, albumin, immunoproteins IgG, IgA, and IgM, and C3C (complement) in cyst fluid more closely correspond to serum than to cerebrospinal fluid values. Values for cyst fluid, cerebrospinal fluid, and serum were determined using albumin ratios in order to compare relative differences between fluids from these three compartments. Our data suggests that: 1) the major protein content of brain tumor cyst fluid is consequential to a transudative process from serum, and 2) that immunoglobulins IgG and IgA are present in higher concentrations in human brain tumor cyst fluids in comparison to IgM concentrations. These studies further question the concept of the brain as an "immunological privileged site", and may be of direct relevance to the investigation of the use of immunotherapeutic modalities as an adjunct after surgical tumor removal.


Changing venous pattern in a hemangioblastoma resembling an arteriovenous malformation. A patient with cerebellar hemangioblastoma underwent two successive cerebral arteriograms within a 48-hour period. The first arteriogram suggested an arteriovenous malformation. The second angiogram showed a dramatic change with nearly total absence of the earlier dilated draining veins. This change was found on examination of the surgical specimen to be caused by venous thrombosis.


Causes of death among construction machinery operators. Causes of death among 2,190 deceased operating engineers identified from the 1967 International Union of Operating Engineers death benefit listing were analyzed to determine if there were conditions occurring with unusual frequency that might be indicative of hazardous conditions in the work environment. The most striking finding was a three-fold excess of fatal accidents occurring at places other than the home or resident institution. The increased risk was evident in each of four regions of the country and at all ages under 65. Significant excesses in deaths from lung cancer and intestinal cancer were seen also. Comparative mortality from lung cancer was directly related to age, with the greatest increase (two-fold) occurring after age 75. There was no evidence of a greater than expected frequency of deaths from non-malignant respiratory disease among operating engineers.


Leptospirosis: a childhood disease.

Leptospirosis: a childhood disease. A diagnosis of leptospirosis was confirmed in nine children who were admitted to St. Louis Children's Hospital during the past 54 months. Epidemiologic, clinical, cultural, and serologic data which were obtained emphasize (1) the high incidence of urban cases; (2) contact with dogs as the most likely source of infection; and (3) that serotypes other than Leptospira icterohaemorrhagiae may produce severe clinical disease. Unusual or previously unreported manifestations of leptospirosis including acalculous cholecystitis, pancreatitis, abdominal causalgia, desquamating skin rashes, and infarction of the extremities which were noted in these children are discussed.


Structural and functional abnormalities of the small intestine due to nutritional folic acid deficiency in infancy. Structural abnormalities and disaccharidase deficiency were demonstrated in biopsies of duodenal mucosa from four infants who had symptoms of failure to thrive and chronic diarrhea associated with a megaloblastic anemia. Goat's milk was the main dietary constituent for each infant for six to eight months prior to presentation. The structural abnormalities consisted of villous blunting, crypt hypertrophy, megaloblastic changes in epithelial cells, and nuclear enlargement. All changes reverted to normal with the addition of folic acid to the diet. This study provides evidence that nutritional folic acid deficiency can cause structural and functional alterations of the small intestine in infants.


Radioimmunoassay of the anticonvulsant agent clonazepam. A simple and specific radioimmunoassay was developed for the determination of the anticonvulsant agent clonazepam directly in plasma without extraction. Antibodies to clonazepam were produced in rabbits after immunization with an immunogen prepared by covalently linking the 3-hemisuccinyloxy derivative of clonazepam to bovine serum albumin. When employing 3H-clonazepam as the tracer, the radioimmunoassay has a limit of sensitivity of 5 ng/ml using a 0.1-ml sample of plasma. The antibodies exhibited a high degree of specificity for clonazepam; no cross-reactivity was observed with its 7-amino and 7-acetylamino metabolites nor with a number of other widely prescribed anticonvulsant agents that might be administered in conjuction with clonazepam. Satisfactory agreement was obtained for the plasma levels of clonazepam in humans when samples were assayed by the radioimmunoassay and an established electron-capture GC technique. By virtue ot its simplicity, the radioimmunoassay offers a distinct advantage to the clinician for monitoring plasma clonazepam levels and the compliance of patients undergoing anticonvulsant therapy with the drug.


Dopaminergic mechanisms in precipitated withdrawal in morphine-dependent rats. Rats were made dependent on morphine by implantation of a pellet and withdrawal was precipitated by the injection of naloxone 72 hours later. Withdrawal was assessed by scoring each of the following signs individually: chewing, licking, teeth chattering, facial tremor, grooming, writhing, diarrhea, weight loss, wet dog shakes, head shakes and hypothermia. The role of dopamine in withdrawal was determined by pretreating the animals with apomorphine or pimozide. Apomorphine in the lower dose range (0.625-1.25 mg/kg) produced a significant decrease in teeth chattering, writhing, weight loss and wet dog shakes. The high dose of apomorphine (2.5 mg/kg) significantly inhibited all features of the withdrawal except writhing and weight loss. Pimozide caused a significant increase in chewing, writhing and head shakes, but only with the highest dose used (0.5 mg/kg). Pimozide (0.5 mg/kg) significantly reduced withdrawal hypothermia, but apomorphine had no effect on this sign except at the highest dose when withdrawal hypothermia was increased.


Absence of formation of brain salsolinol in ethanol-dependent mice. The in vivo biosynthesis of tetrahydroisoquinoline (TIQ) alkaloids has been shown to occur during acute and chronic ethanol administration under experimental conditions which increase aldehyde precursors and reduce TIQ metabolism. The TIQs have been postulated to mediate some of the effects of ethanol, although their occurrence under conditions of physical dependence has not been reported. The purpose of this investigation was to determine whether in vivo TIQ formation would occur in brains of ethanol-dependent animals in the absence of specific experimental manipulations, and their role, if any, in the development of physical dependence. Mice were subjected to ethanol vapors for 6 days (14 mg/1 in inspired air on day 1, increasing to 20-30 mg/l on day 6), and were then either evaluated for withdrawal convulsions on handling or sacrificed and their brains analyzed for salsolinol (the acetaldehyde-dopamine TIQ) by gas chromatography/electron capture (assay sensitivity: 8 ng/g of tissue). No evidence could be obtained for in vivo formation of salsolinol in individual whole brains, pooled whole brains or dopamine-rich areas of pooled brains obtained from ethanol-dependent mice even under conditions where blood ethanol levels reached 7 mg/ml. In view of the low pharmacological potency of salsolinol, it is unlikely that this alkaloid plays a significant role in the effects of ethanol in mice.


Neurochemical aspects of ethanol dependence and withdrawal reactions in mice. Mice were made physically dependent on ethanol by a 3-day period of alcohol inhalation with small daily injections of pyrazole. During this treatment the concentrations of norepinephrine, dopamine and 5-hydroxytryptamine were increased in brain with concomitant decrease in gamma-aminobutyric acid, RNA and DNA. However, the monoamine concentrations showed complete regression to normal levels at the time of maximal withdrawal seizure when GABA level was still elevated above the control values. Brain RNA and DNA concentrations remained low at this period. During the recovery phase, the pattern of neuronal components was almost the same as was observed at maximal withdrawal seizures. Pyrazole by itself did not produce significant changes in concentrations of the neuronal components of brain.


The use of R-R interval and difference histograms in classifying disorders of sinus rhythm. A data reduction technique is presented which enables the patterns of cardiac rhythm inherent in a long-term ECG recording to be described in a concise fashion. The method consists in combining an R-R interval difference histogram, which gives information about the detailed structure of cardiac rhythm patterns, with an R-R interval histogram, which concisely describes the trends in heart rate over a period. Example of the histogram patterns corresponding to some common cardiac rhythms are presented. The histograms can be treated mathematically to yield information which is useful in assessing the long-term effects of drugs and in classifying disorders of cardiac rhythm.


Association of cancer sites with tobacco and alcohol consumption and socioeconomic status of patients: interview study from the Third National Cancer Survey. From personal interviews obtained for 7,518 incident cases of invasive cancer from the population-based Third National Cancer Survey, the quantitative lifetime use of cigarettes, cigars, pipes, unsmoked tobacco, wine, beer, hard liquor, and combined alcohol were recorded, as well as education and family income level. In an initial screening analysis of these data, Mantel-Haenszel 2 X 2 contingency tabulations and multiple regression analyses were used to compare each specific cancer site with controls from other sites to test for associations with the "exposure variables." Significant positive associations with cigarette smoking were found for cancers of the lung, larynx, oral cavity, esophagus, stomach, pancreas, bladder, kidney, and uterine cervix. Other forms of tobacco were associated with cancers of the oral cavity, larynx, lung, and cervix. Consumption of wine, beer, hard liquor, and all combined showed positive associations with neoplasms of the oral cavity larynx, esophagus, colon, rectum, breast, and thyroid gland. College educaton and high income both showed positive associations with cancers of the breast, thyroid gland, uterine corpus, and melanomas in males. These same indicators of high socioeconomic status showed inverse associations with invasive neoplasms of the uterine cervix, lung, lip-tongue, and colon in females. College attendance (but not income) showed an inverse association with stomach cancer and positive association with pancreatic cancer in males. Still other tumor sties showed "suggestive" associations with each of these exposure variables. In the analyses producing these results, age, race, sex, smoking, drinking, education, income, parity, foreign birth, marital status, and geographic location were used as stratification variables separately or in combination when appropriate to assess and control for their potentially confounding affects and to examine results in different strata to assess interaction.


Fractured pelvis with avulsion of the female urethra. Injuries to the female urethra, except for meatal straddle injuries and those associated with instrumentation, are rarely seen with pelvic trauma. To our knowledge this is the first case of traumatic complete disruption of the female membranous urethra reported in the English literature.


Carcinogenesis in the pancreas. I. Long-term explant culture of human and bovine pancreatic ducts. Bovine and human pancreatic ductal explants were maintained in long-term culture. Bovine ducts were cultured for 85 days, whereas human ducts have been cultured for up to 60 days. The explants all maintained good ultrastructural preservation for these periods and also incorporated radioactive precursors into protein, RNA, and DNA. Uncultured ducts, which were fixed for electron micrsoscopic study, demonstrated classical signs of reversible cell injury (dilated endoplasmic reticulum and swollen mitochondria), and cultured explants did not. However, the cultured tissues did show sublethal alterations such as the formation of numerous autophagic vacuoles and residual bodies and the accumulation of large lipid droplets. This study demonstrated the feasibility of maintaining pancreatic ducts in long-term explant culture, which enables them to be used in experimental studies involving chemical carcinogens and in structural and functional studies.


The gross and histopathologic lesions of maignant catarrhal fever in three captive sika deer (Cervus nippon) in southern Ontario. The gross and histopathologic lesions of three captive sika deer (Cervus nippon) with malignant catarrhal fever are described. Lesions included those of the head and eye form and the more commonly described peracute form. One deer had been exposed to a wildebeeste (Connochaetes gnou) and the other two to domestic sheep.


[Clinical studies on clindamycin-2-phosphate in the field of surgery (author's transl)]. Twenty eight patients were treated with parenteral clindamycin-2-phosphate in the field of surgery, and good response was obtained in a series of superficial soft tissue infection, especially caused by staphylococci, with a daily dose of 300 mg. Serum level and urinary excretion were also investigated in four healthy male volunteers.


Changes in hepatic collagen metabolism in rats produced by chronic ethanol feeding. The effect of ethanol feeding for a period of 6 months on parameters of hepatic collagen metabolism was studied in the rat. Ethanol feeding resulted in small increases in the fibrous and ground substance components of hepatic collagen as measured by increases in collagen-bound hydroxyproline and hexosamine, respectively. Liver histology revealagen proline hydroxylase and the incorporation of labeled proline into collagen by liver slices, both of which are associated with collagen synthesis, were not changed. Ethanol feeding resulted in increases in the concentration of protein and deoxyribonucleic acid in the Kupffer cells, but in no changes in collagenase activity. An increase in collagen degradation was suggested, however, by the increase in the urinary excretion of hydroxyproline and glycosaminoglycans found after 2 and 6 months of ethanol feeding, respectively. This study demonstrates that fatty infiltration of the liver in the rat, after prolonged ethanol feeding, is associated with increased deposition of chemically detectable collagen and evidence of increased collagen degradation, although no significant changes in parameters associated with hepatic collagen synthesis were found.


A reexamination of the role of heredity in stuttering. In the light of recent breakthrough in the study of schizophrenia indicating a far stronger genetic factor than has been previously suspected, the role of heredity in stuttering is critically reexamined. Present knowledge of the role of heredity in stuttering springs from four principal data sources: (1) studies of familial incidence; (2) spontaneous recovery studies; (3) twin studies; and (4) studies of parental disfluency. It is hypothesized that the 4:1 sex ratio in stuttering may be the product of selective genetic factors. Critical examination of earlier and more recent studies leads to substantial evidence that a familial predisopsing factor exists in about 25% cases of stuttering.


Base-10 programmed stimulation: task specification, scoring, and plotting performance in aphasia therapy. This report explains and advocates the applicability of time-series designs to document the effects of therapy with aphasic subjects; it describes a measurement system that contributes to organization of therapy, task specification and scoring, and graphic display of change in speech and language behaviors. Examples of patient performances illustrate the application of Base-10 Programmed Stimulation to therapy with aphasic persons.


Auditory symptoms associated with herpes zoster or idiopathic facial paralysis. Auditory symptoms (hyperacusis, tinnitus, decreased hearing) have long been recognized to accompany herpetic or idiopathic facial paralysis. Twenty-nine percent of 1,080 patients with idiopathic facial paralysis and 37 percent of 172 with herpes zoster oticus facial paralysis had auditory symptoms. Abnormal related sensori-neural hearing loss was documented in only 11 of these 377 patients with auditory complaints. All of the 11 had a diagnosis of herpes zoster oticus. Sensori-neural hearing loss occurs in only about 6.5 percent of patients with herpes zoster facial paralysis, and no confirmed case of such loss in idiopathic facial paralysis has been reported. In patients presenting with sensori-neural hearing loss accompanying facial paralysis believed to be idiopathic, herpes zoster should be suspected even in the absence of vesicles. Factors favorable for recovery of auditory function include age 64 years or younger, mild initial hearing loss, a cochlear pattern of hearing loss, and absence of vertigo. Recovery of auditory function does take place; however, a high-tone sensori-neural loss may persist except in younger patients.