Use of fecal streptococci as indicators of pollution in soil.

The survival, recovery, and identification of Streptococcus isolates from soil was investigated by (i) examination of survival in soil under different moisture and temperature conditions, (ii) evaluation of media combinations for recovering fecal streptococci from soil, and (iii) partial identification of isolates from diverse habitats. Cool, moist conditions prolonged the survival of Streptococcus faecalis in soil for at least 12 weeks, whereas freezing was lethal, with the populations being reduced up to 95% when several freeze-thaw treatments occurred. Media evaluations indicated that both the efficiency of recovery and enumeration of the fecal streptococci from soil can be influenced by the combination of media used. Taxonomic data revealed a need to develop procedures to differentiate between isolates of fecal origin and plant-derived streptococci that possess many of the cultural reactions of S. faecalis. It was found that recent fecal isolates exhibited a much greater incidence of multiple antibiotic resistance than soil or vegetation isolates, and this characteristic, coupled with the use of enterococci as indicators of fecal contamination in soil systems, is discussed.


Use of extractable adenosine triphosphate to estimate the viable cell mass in dental plaque samples obtained from monkeys.

The viable cell mass in plaque samples obtained from monkeys was estimated by determining the concentration of extractable adenosine triphosphate (ATP), and total cell mass was estimated by measuring the protein content. The results were expressed in terms of the specific ATP and protein contents of Streptococcus sanguis. The viable counts estimated by these techniques were comparable to or exceeded viable counts obtained by other investigators using conventional bacteriological methods.


Osmotically induced increase in thermal resistance of heat-sensitive, dipicolinic acid-less spores of Bacillus cereus Ht-8.

Thermal resistance in heat-sensitive, dipicolinic acid (DPA)-less spores of Bacillus cereus Ht-8 heated in sucrose solutions increased at and above a concentration of 2 M sucrose. The decimal reduction times at 75 degrees C for spores heated in 0.0, 1.8, 2.2, and 2.6 M sucrose were 2.0, 2.8, 4.5, and 12 min, respectively. Maltose, fructose, and glucose increased heat resistance above that observed in water but did not elevate resistance to the level observed with sucrose at the same osmolality. Cation-induced loss of thermal resistance in chemically sensitized spores was reversed in the presence of sucrose. Spores germinated in brain heart infusion were resistant when heated in sucrose. In the presence of sucrose, spores exhibited an increase in optical density at 700 nm. Electron micrographs of the DPA-less spores suspended in 2.2 M sucrose revealed a shrinkage of outer coats and exosporium membranes. The results suggested that the osmotic property of sugars increased thermal resistance in DPA-less spores. The osmotic pressure exerted by sugars may be similar to the pressure that usually exists within the cortex of normal spores containing DPA and may cause the dehydration of the protoplast and the consequent thermal resistance. The role of dehydration and the nonessential nature of DPA for thermal resistance in spores were confirmed.


Improved tube for nasojejunal feeding in low birthweight infants.

A simple tube for nasojejunal feeding is described. 22 infants with birthweight less than 1500 g and 25 infants 1500--2000 g were fed during the first weeks of life. The tube elimintes many of the difficulties previously encountered with nasojejunal feeding.


Fat embolism in infancy after intravenous fat infusions.

Four cases of fat embolism are described in infants receiving prolonged intravenous infusion of fat (Intralipid 20%). This therapeutic complication has been termed 'fat overloading syndrome' but bears a clinical similarity to post-traumatic fat embolism. These 4 cases are the first to be recorded in infancy, and with histopathological proof of fat embolism. These 4 cases are the first to be recorded in infancy, and with histopathological proof of fat embolism. Transient high rates of infusion of Intralipid appears to be a factor in the aetiology of the condition.


Combined test of anterior pituitary function in children.

A combined test of anterior pituitary function was used on 21 normal children. It shows that a comprehensive evaluation can be made of growth hormone, thyroid stimulating hormone, adrenocorticotrophic hormone, and gonadotrophin reserve. Simultaneous assessment of those peripheral glands associated with the anterior pituitary is possible by the measurement of thyroid hormones, cortisol, and gonadal steroids. The procedure can be completed in 4 hours with minimum inconvenience and distress to the child.


Loss of energy during continuous infusions of breast milk.

During 9 experiments simulating continuous intragastric infusion of human breast milk, variations in milk energy content of up to 24% occurred, the samples taken at the end of 8-12 hours' infusion being particularly energy-rich compared with earlier samples. Burettes should be regularly agitated during infusions of human milk.


Air embolism occurring as a complication of central venous catheterization.

Two patients with air embolism (one fatal) which occurred during intravenous feeding with a central venous catheter are reported. The connection between intravenous administration sets and such catheters is shown to be a danger area which requires further technological improvement. The world literature concerning this hazard is reviewed.


Ten years experience with intravenous hyperalimentation and inflammatory bowel disease.

A retrospective analysis was conducted on 74 patients with inflammatory bowel disease who were treated with intravenous hyperalimentation at the Hospital of the University of Pennsylvania between the years 1967-1976. Intravenous hyperalimentation can ameliorate the inevitable protein-calorie malnutrition present in patients with inflammatory bowel disease. Combined with complete bowel rest, intravenous hyperalimentation can effectively function as the primary treatment or as an adjunct to the surgical management of the complications of inflammatory bowel disease. Intravenous hyperalimentation can be safely administered to these severely ill patients, almost certainly improving survival rates in the patients treated.


Electro-acupuncture analgesia in monkeys: a behavioral and neurophysiological assessment.

Monkeys were initially trained to press a lever to escape noxious electrical stimulation applied to one leg. They were then presented with electrical stimuli consisting of 40 successive intensity increments beginning at zero intensity and with random interstimulus intervals (ISIs). A lever press, initiated by the monkey during presentation of a stimulus, immediately terminated stimulation of the leg, triggering another sequence of the same increasing stimuli with a new series of ISIs. After establishment of a stable threshold measure of lever pressing, 45 min of electro-acupuncture was applied to traditional points on both legs. The results show that electro-acupuncture dramatically increased (300-400 %) the lever pressing threshold for approximately 20 min after electro-acupuncture stimulation was terminated. In acute monkey preparations, spike activity of single neurons in N. Parafascicularis of the thalamus, evoked by noxious electrical stimulation to the sciatic nerve, was recorded. The evoked activity of only nociceptive specific neurons was either eliminated or markedly reduced for at least one hour following 45 min of electro-acupuncture applied to traditional points on both legs. The observations made by these two experimental approaches suggest that electro-acupuncture exerts an analgesic effect on artificially induced nociception.


Massive proteinuria in light chain disease.

A 33-year-old man with overall renal function in the lower normal range had daily excretion in the urine of between 31 and 70 gm of protein composed entirely of free monoclonal K light chains. K light chains were also present in the serum. Serum protein electrophoresis and findings on bone marrow and lymph node biopsy were diagnostic of light chain disease. Amyloid was absent from renal tissue. General clinical improvement and almost total disappearance of protein from the urine followed treatment with phenylalanine mustard.


Correction of taste abnormality of malignancy with intravenous hyperalimentation.

A complete remission of widely metastatic bladder carcinoma was obtained with chemotherapy in a 41-year-old man. Severe persistent anorexia led to a 33% weight loss. The anorexia correlated with an elevated sucrose recognition threshold. Following 16 days of intravenous hyperalimentation, taste sensation returned to normal, anorectic symptoms cleared, and weight gain and positive nitrogen balance resulted. There was no stimulation of tumor growth. Restoration of nutritional deficits with intravenous hyperalimentation can improve taste function and appetite so that adequate oral alimentation can be tolerated.


Hyponatremia of hypothyroidism. Appropriate suppression of antidiuretic hormone levels.

A hypothyroid, 72-year-old woman with idiopathic hypopituitarism manifested severe hyponatremia, plasma hypoosmolality, and inappropriately elevated urine osmolality suggestive of a syndrome of inappropriate antidiuretic hormone secretions. The hyponatremia did not respond to demeclocycline hydrochloride, and antidiuretic hormone (ADH) levels measured by a specific radioimmunoassay were appropriately suppressed. Subsequent replacement therapy with levothyroxine sodium resulted in correction of the hyponatremia. Thus, both direct assay as well as hormone blockade failed to show an action of ADH in mediating the water retention.


Liver disease and vasculitis in a patient taking cromolyn.

Hypersensitivity reactions to cromolyn sodium occur rarely. On several occasions they have been associated with peripheral eosinophilia and granulomatous inflammation. Liver disease has not been reported previously as a complication of inhaled cromolyn. We describe here a woman in whom marked peripheral eosinophilia, liver disease, and systemic vasculitis developed while taking cromolyn and resolved or improved on discontinuation of the drug and treatment with corticosteroids. The liver disease was similar to primary biliary cirrhosis except that marked eosinophilic infiltration and granulomas were present initially. Studies of the patient's serum for binding of carbon 14-labeled cromolyn, the skin for deposits of the drug, and the circulating lymphocytes for stimulation by cromolyn failed to demonstrate any abnormalities. However, the elevated IgG and IgM levels, the positive rheumatoid factor and antimitochondrial antibody, and the reduced serum complement, which returned to normal on discontinuation of the drug therapy, suggests that immunologic mechanisms may have played a role in the pathogenesis of this patient's illness.


Variations in short term products of inorganic carbon fixation in exponential and stationary phase cultures of Aphanocapsa 6308.

Aphanocapsa 6308 metabolizes both NaHCO3 and Na2CO3. The short term incorporation (5-s) metabolic pattern and the patterns of incorporation of bicarbonate for exponential versus stationary phase cultures differ, however. Cells were equilibrated for 10 min in air and distilled water prior to injection of either NaH14CO3 at pH 8.0, or Na214CO3 at pH 11.0. Hot ethanol extracts were analyzed via paper chromatography and autoradiography for products of CO2 fixation. At 5 s, malate (51.5%) predominates slightly as a primary bicarbonate fixation product over 3-phosphoglycerate (40.3%); 3-phosphoglycerate is the primary product of carbonate fixation. At 60 s, the carbonate and bicarbonate labelling patterns are similar. Cells in stationary phase fix in 5 s a greater proportion of bicarbonate into malate (36% vs. 14% for 3-phosphoglycerate) than do cells in exponential growth. Likewise, 60 s incorporations show a large amount of bicarbonate fixed into aspartate (30.9%) in stationary phase cells over that of exponential phase (11.6%). These data suggest an operative C4 pathway for purposes not related to carbohydrate synthesis but rather as compensation for the incomplete tricarboxylic acid cycle in cyanobacteria. The enhancement of both aspartate fixation and CO2 fixation into citrulline in stationary phase correlates with an increase in cyanophycin granule production which requires both aspartate and arginine.


Metabolic products of microorganisms 167. Cyclopaldic acid from Aspergillus duricaulis. 1. Production, isolation and bioloical properties.

In the course of a screening for new metabolites from fungi we isolated a substance with antimicrobial activity from cultures of Aspergillus duricaulis (CBS 481.65) (Tü 679). It was antagonized by putrescine, spermidine, spermine, arginine, citrulline, lysine, ornithine, in higher concentration by aspraagine and glutamine too. The effect of ethylenediaminetetraacetate on the susceptibility of Streptomyces viridochromogenes (Tü 57) and Bacillus subtilis ATCC 6051 to this antibiotic has been studied. The substance was characterized and identified as cyclopaldic acid.


Determination of kallidinogenase activity by radial immunodiffusion methods.

An attempt has been made to elaborate a method allowing determination of kallidinogenase obtained from the pancreas of slaughtered animals (pigs), by means of radial immunodiffusion method. As the test substance (antigen) a pharmaceutical preparation Dilminal D has been used. The immune serum against the preparation administered was obtained from rabbits. This serum was used to saturate plates of agar gel. Little pigs cut in the gel were filled with various concentrations of the preparation (antigen). A linear correlation between the selected concentrations of the preparation was obtained. This method may find application in determining preparations of kallidinogenase (from pig pancreas) with titer value given in different units.


Stroke rehabilitation: a reconsideration of some common attitudes.

Some common, yet erroneous, attitudes and perceptions about stroke still persist. These warrant reconsideration: (1) benefits of stroke rehabilitation (including validity of its basis, life expectancy, adequacy in nursing homes, outcome prediction, cost benefits, and vocational outcome); (2) gait training (including evaluation methods, gait patterns, hand supports, sensory deficits, and types of braces); (3) effects of training on regaining balance; (4) spasticity (as a negative factor, enhancement by spring-action brace, benefit of inhibition training, and importance of antispasmotic drugs); (5) danger of early activity; (6) depression; (7) effects on patients' sexuality; (8) effects of communication impairments on learning abilities as well as effectiveness of speech therapy; (9) application of neurphysiological principles (regarding decreasing synaptic resistance, applications of principles from cerebral palsy training, and benefits of training for percept-concept-motor function deficits); and (10) research including reliability of past reports and paucity of facilities for new research.


Reciprocal changes in primary and secondary optokinetic after-nystagmus (OKAN) produced by repetitive optokinetic stimulation in the monkey.

In six rhesus monkeys (Macaca mulatta) the effect of repetitive periods of whole-field optokinetic stimulation upon the different phases of optokinetic after-nystagmus (OKAN) was studied. The precedings optokinetic stimulus consisted of rotating a striped cylinder around the stationary monkey. Experiments were performed on up to 8 successive days. The results demonstrate that OKAN I and OKAN II are affected in a reverse manner by repeated optokinetic stimulation: The duration of OKAN I strongly decreases, whereas OKAN II increases in duration and intensity. The mechanisms under lying the generation of OKAN I and OKAN II and the role of OKAN II in the habituation process are discussed.


Endotoxin-challenged monkeys and rats.

Studies in our laboratory with both the monkey and the rat showed that, after three hours of endotoxemia, there was a significant decrease in the number of circulating platelets, total hemolytic complement (CH 50 units), and blood serotonin (5-HT) levels. Administration of dexamethasone sodium phosphate in the clinical dose range at the time of endotoxin challenge significantly attenuated the decrease in blood 5-HT levels when compared to the untreated groups in both the monkey and the rat experiments. In the monkey, CH 50 units remained at a higher level when dexamethasone was administered; however, the difference between the treated and untreated groups was not statistically significant. The number of circulating white blood cells and platelets did not appear to be significantly altered by corticosteroid treatment. It is suggested that glucocorticoids may interfere with lipopolysaccharide-induced alterations in complement components or factors regulating hemostasis that influence platelet 5-HT release.


Management of septic thrombosis of the inferior vena cava caused by Candida.

Septic thrombosis of central veins is rarely diagnosed during life and nearly always proves fatal. We have recently successfully treated a patient with a 75% body surface burn in whom septic thrombosis of the inferior vena cava developed associated with high-grade candidemia as a complication of parenteral nutrition. Signs of venous thrombosis and candidemia persisted after catheter removal. Prompt and intensive therapy with amphotericin B, monitored by fungicidal assays of serum, resulted in cure. Generous hydration and directed supplementation of sodium bicarbonate permitted us to administer a large total dose of amphotericin over a relatively brief period of time with no nephrototoxic effect whatsoever. Septic central venous thrombosis mandates a pharmacologic approach to therapy similar to that used for infective endocarditis, with the addition of anticoagulation. Should sepsis prove refractory to this program of it pulmonary embolization occurs, operative intervention is indicated despite the high risks involved.


Squirrel monkey retrovirus: electron microscopy of a virus from New World monkeys and comparison with Mason-Pfizer monkey virus.

The ultrastructural morphogenesis of squirrel monkey retrovirus (SMRV) and Mason-Pfizer monkey virus (MPMV) growth in cell culture were compared. Both viruses develop by a process that begins with the formation of intracytoplasmic A particles which are then enveloped at the plasma membrane during budding. SMRV also develops as a crescent-shaped nucleoid beneath a bulging plasma membrane, a development characteristic of type C oncornaviruses. Free extra-cellular mature SMRV was generally round with a centrally located electron-dense nucleoid enclosed by the viral envelope. In contrast, mature MPMV had a tubular-shaped nucleoid. Negative stained preparations of both viruses yielded head-tail forms with surface projections. By uranyl acetate/critical point drying, SMRV particles were usually round with an eccentric electron-dense nucleoid enclosed by the viral envelope, whereas MPMV particles were round and contained an electron-dense bar-shaped nucleoid. These morphological observations indicate that SMRV more closely resembles MPMV, presently the only member of genus oncornavirus type D, than other retroviruses species. However, since SMRV can be morphologically, biochemically, and immunologically distinguished from MPMV, it represents a new species within genus oncornavirus type D.


[Comparative morphologic features of the retinas of insectivores and primates].

By means of scanning electron microscopy and by neurohistological methods with subsequent quantitative treatment of the data, it is demonstrated that the retina of the hedgehog differs from that of the green monkey. In the former, the density of photoreceptors convergence is higher, that corresponds to the night mode of life of the hedgehog Monotony in the cells of all the layers, lack of photoreceptor differentiation according to the form of their external segments corresponds to general low level in the organization of the hedgehog nervous system. The retina of the hedgehog possessing the above mentioned structural peculiarities can be considered as a model reflecting the organization in the peripheral portion of the optic analyzer of the original forms in Mammalia placentalia.


EEG patterns at the time of focal seizure onset.

We studied the electroencephalographic pattern at the time of focal seizure onset in 41 patients. Progressive rhythmic EEG activity was characteristic of spontaneous focal seizures in man. Focal hypersynchrony was present in 49% of cases and served as an accurate localizing indication of cortical irritability. Interictal sharp or spike discharges were absent in 34% of cases. When present, spike and sharp discharges characteristically attentuate at the onset of rhythmic ictal activity. Recognition of the pattern of rhythmic ictal transformation is often essential for the electroencephalographic diagnosis of focal seizures.


Involvement of peripheral nerve and muscle in Fabry's disease. Histologic, ultrastructural, and morphometric studies.

Light microscopic and ultrastructural studies of biopsy specimens from the sural nerve and the gastrocnemius muscle in a patient with Fabry's disease showed accumulation of lipids in endothelial and perithelial cells of the vessel walls. In addition, the peripheral nerve exhibited deposition of lipids in the perineurial cells, occasionally in unmyelinated and myelinated axons, and infrequently in Schwann cell cytoplasm. In the muscle biopsy specimen, stored lipid was found in the sarcoplasm. Quantitative histologic studies showed loss of large unmyelinated and thin myelinated nerve fibers. Excruciating pain and loss of sweating, characteristic of this disorder, may result from loss of these fiber categories. The peripheral neuropathy is probably secondary to perikaryal deposition of lipid as described previously in the literature.


Impedance audiometry in the squirrel monkey. Effect of middle ear surgery.

The two principal components of impedance audiometry, the tympanogram and acoustic reflexes, were measured preoperatively and postoperatively in seven squirrel monkeys subjected to experimental middle ear surgery. A probe-tone frequency of 1,200 Hz yielded both tympanograms and acoustic reflex thresholds roughly comparable to analogous data in humans at a probe-tone frequency of 220 Hz. Both tympanograms and acoustic reflex thresholds could be obtained reliably and with comparative ease. In general, postoperative data were modified in the expected fashion by the experimental middle ear surgery. Impedance audiometry appears to be a worthwhile and useful technique for monitoring middle ear status in the squirrel monkey.


Symptomatic ossification of the pterygomandibular raphe.

A case of symptomatic ossification of the pterygomandibular raphe, believed to be the first reported case of such an entity, was relieved by surgical removal of the ossified tissue. Ossification of the pterygomandibular raphe should, in the future, be considered in the differential diagnosis in cases of obscure lateral pharyngeal pain.


Hearing loss and cochlear pathology in the monkey (Macaca) following exposure to high levels of noise.

Eight Old World monkeys were exposed 8 h daily for 20 days to octave-band noise having center frequencies from 0.5--8 kHz at levels of 117--120 dB SPL. Two additional animals received exposures to wide-band, 120-dB SPL noise on the same schedule, and one animal was exposed to the 2-kHz octave band for 40 h continuously. Behavioral audiograms were measured throughout exposure and during a 1-month recovery period. Following recovery, the animals were sacrificed and their ears examined histologically. Monaural audiograms are presented showing initial and final TTS and PTS measured at the end of the recovery period. These are compared with complete cytocochleograms for each ear. Hair cell loss was generally restricted to the outer rows, and was reasonably well correlated with pattern of hearing loss. Some cell loss, including inner hair cells, was found in the extreme basal turn, usually without associated high-frequency hearing loss. The relationships between exposure frequency, hearing loss, and locus of cochlear pathology are discussed, as are changes in TTS during exposure.


Acute recreational injuries.

The injuries which occur in recreation have the effect of eliminating participation for a period and also in many cases lead to a cessation of physical activity. The loss of well being as a result of idleness is a price that today's inactive world cannot afford to pay.


The New South Wales mastitis control program. 3. The effect on production and economics.

A mastitis control program carried out in 35 commercial dairy heards over three years increased the milk produced by 343 kg per cow per year and the amount of butterfat by 15.8 kg per cow per year, giving a net benefit of $16.10 per cow per year by the end of the third year. The net benefit is considerably understated since benefits other than the increase in milk production were not quantified. The overall disposal rate of 33 herds showed a drop from 19.3% in year 1 to 13.6% in year 3. An analysis of culling records showed that as the severity of mastitis increased so did cow age.


Drosophila hemolymph proteins: purification, characterization, and genetic mapping of larval serum protein 2 in D. melanogaster.

Three of the major protein species present in the hemolymph of Drosophila melanogaster larvae just prior to pupation are absent from second instar larvae but accumulate rapidly during the third instar. This article describes the purification and characterization of one of these, larval serum protein (LSP) 2, using an immunological assay. It is a homohexamer of molecular weight about 450,000, with a polypeptide molecular weight of 78,000--83,000. Fast and slow electrophoretic variants of this protein map between the markers vin and gs, at 36--37 on chromosome 3.


Isolation of turkey immunoglobulin-A.

Trukey biliary and gut IgA were isolated and monospecific antisera were prepared in rabbits. Isolation was accomplished by filtration of precipitated globulins on Sephadex G-200 followed by step-wise-elution DEAE ion-exchange chromatography. Using an immunodiffusion procedure, IgA was detected in turkey serum, saliva, lacrimal secretions, bile, gut washings, and tracheal washings. No IgA was detected in hatching-poult serum, egg white, or yolk. Two forms of IgA (high and low molecular weight) were detected in different body fluids.


Microsporidian infection in the pied peach-faced lovebird (Agapornis roseicollis).

A microsporidian infection was diagnosed in a pied peach-faced lovebird (Agapornis roseicollis) which had died after an illness of 2 days. The parasite was observed in the liver, kidney, and small intestine. Lesions were most severe in the liver, with numerous organisms adjacent to areas of hepatic necrosis. Although the organisms were seen within renal tubules and within the intestinal mucosa, inflammatory lesions in those sites were minimal. The microsporidian nature of the parasite was confirmed by morphologic studies. The genus of the organism was not determined.


Control of drosopterin synthesis in Drosophila melanogaster: mutants showing an altered pattern of GTP cyclohydrolase activity during development.

The reaction catalyzed by GTP cyclohydrolase is the first unique step of pteridine biosynthesis in Drosophila melanogaster and is therefore likely to be an important control point. GTP cyclohydrolase activity varies during development, showing two distinct peaks of activity--one at pupariation and a much larger peak at emergence. Most of the early pupal enzyme is located in the body region, whereas in late pupal and early adult life most of the activity is found in the head. Mixing experiments indicate that developmental changes in activity are not due to changes in the level of a direct effector of GTP cyclohydrolase. The mutants raspberry and prune show an increased GTP cyclohydrolase activity at pupariation relative to wild type, but a decreased enzyme activity at emergence. The changes in GTP cyclohydrolase activity are reflected in changes in pteridine levels in these mutants. Several lines of evidence suggest that neither locus is the structural gene for GTP cyclohydrolase. The raspberry and prune gene products may play a specific role in regulating GTP cyclohydrolase activity during development.


Leucocytozoon smithi: persistence of gametocytes in peripheral turkey blood.

Ten-week-old Broad-breasted White turkeys were exposed for 2 weeks to a natural infection of Leucocytozoon smithi in Marlboro and Sumter Counties in South Carolina. The birds were then returned to Clemson, South Carolina, and housed in pens proofed against black flies (Simulium sp.) to prevent any reinfection. Blood smears, stained with Giemsa's stain, were prepared at regular intervals to determine the presence of gametocytes of L. smithi after a single natural infectional exposure. Gametocytes were observed in the blood smears for 13 consecutive months, after which the experiment was terminated.


Nonsecreting myeloma variants with heavy-chain carbohydrate deficiencies.

Light- and heavy-chain synthesis was studied in six previously isolated S194-2 mouse myeloma variant lines and in the parent from which they were derived. Serological data and comparative analysis of the cyanogen bromide fragments obtained from variant and parent intracellular immunoglobulin showed that five variants which failed to secrete detectable amounts of IgA synthesized both heavy- and light-chain subunits. Whereas at least two heavy-chain populations were resolved in the parent line, one containing carbohydrate and one devoid of carbohydrate, only the heavy-chain fraction devoid of carbohydrate was detected in the variant lines. The correlation between carbohydrate deficiencies on the heavy chains and lack of immunoglobulin secretion in five independent subclones is discussed in terms of possible primary lesions and the role of carbohydrate in secretion.


Extensive homologies between the methylated nucleotide sequences in several vertebrate ribosomal ribonucleic acids.

The methylated nucleotide sequences in the rRNA molecules of the following vertebrate cultured cells were compared: human (HeLa); hamster (BHK/C13); mouse (L); chick-embryo fibroblast; Xenopus laevis kidney. In each species the combined 18S, 28S and 5.8S molecules possess approx. 110-115 methyl groups, and the methylated oligonucleotides released after complete digestion of the rRNA by T1 ribonuclease encompass several hundred nucleotides. "Fingerprints" of the three mammalian methyl-labelled 18S rRNA species were qualitatively indistinguishable. "Fingerprints" of digests of 28S rRNA of hamster and mouse L-cells were extremely similar to those of HeLa cells, differing in one and three methylated oligonucleotides respectively. "Fingerprints" of methyl-labelled rRNA from chick and Xenopus strongly resembled those of mammals in most respects, but differed in several oligonucleotides in both 18S and 28S rRNA. At least some of the differences between "fingerprints" appear to be due to single base changes or to the presence or absence of methyl groups at particular points in the primary sequence. The findings strongly suggest that the methylated-nucleotide sequences are at least 95% homologous between the rRNA molecules of the two most distantly related vertebrates compared, man and Xenopus laevis.


Purification and characterization of the class-II D-fructose 1,6-bisphosphate aldolase from Escherichia coli (Crookes' strain).

A new form of the class-II D-fructose 1,6-bisphosphate aldolase (EC 4.1.2.13) of Escherichia coli (Crookes' strain) was isolated from an extract of glycerol-grown bacteria. It has a higher molecular weight (approx. 80000)than previous preparations of the enzyme and closely resembles the typical class-II aldolase from yeast in size and amino acid composition. On the other hand, its kinetic behaviour is not typical of a class-II aldolase. The enzyme has no requirement for thiol compounds either for stability or activity, added K+ ions have no effect, and the optimum pH for the cleavage activity is unusually high. The class-II enzymes from the prokaryote E. coli and the eukaryote yeast show no immunological identity. However, the similarity of their structures suggests that they have evolved from a common ancestor.


Purification of rat fibrinogen and its constituent chains.

Rat fibrinogen was purified from rat plasma by using lysine-Sepharose chromatography, repeated precipitation with 25%-satd. (NH(4))(2)SO(4) and gel chromatography on Sepharose 6B. To minimize proteolytic activity, rats were injected intravenously with Trasylol before bleeding and the collected blood was treated with Trasylol and di-isopropyl phosphorofluoridate. A 95%-clottable preparation was obtained in 70-75% yield; it proved to be free of factor XIII and plasminogen. It showed a single band on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis and on disc electrophoresis in 8m-urea. Alanine was the only detectable N-terminal amino acid. After reduction and modification of the thiol groups, the material could be separated into three distinct chains (Aalpha, Bbeta and gamma) by pore-limit polyacrylamide slab-gel electrophoresis in the presence of sodium dodecyl sulphate. The amino acid compositions of the whole fibrinogen and of the separated modified chains were determined. The molecular weights were 61000, 58000 and 51000 for Aalpha-, Bbeta- and gamma-chains respectively. Our results for the chains are in contrast with previous reports on rat fibrinogen [Bouma & Fuller (1975) J. Biol. Chem.250, 4678-4683; Stemberger & Jilek (1976) Thromb. Res.9, 657-660], in which no separation between Aalpha- and Bbeta-chains was achieved on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis for 3h. Evidence is presented that this is probably due to Aalpha-chain degradation as a result of incomplete inhibition of proteolytic enzymes during the purification. Complete inhibition of proteolytic activities is essential in all steps of the present purification procedure.


Demineralization in organic solvents by alkylammonium salts of ethylenediaminetetra-acetic acid.

1. Insoluble salts of alklaine earth metals (e. g. calcium phosphate) dissolve in non-aqueous solvents containing EDTA salts of organic cations (e.g. tetraethylammonium). 2. Bone is efficiently demineralized in ethanolic trimethylammonium EDTA. Retention in the matrix of water-soluble materials (e.g. proteoglycan) is much better than in standard aqueous EDTA demineralization procedures.


Affinity chromatography of the cellulase system of Trichoderma koningii.

A procedure involving affinity chromatography on cellulose was developed for separating enzymic components of the cellulase complex. Cellobiase, carboxymethylcellulase, component C2 and cellobiohydrolase are adsorbed with increasing tenacity, and released, as highly purified components, as the ionic strength of the eluent is decreased.


Recovery from vitamin B-12-induced unbalanced growth. The shortened cell cycle and the deoxyribonucleoside triphosphate pools.

The deoxyribonucleoside triphosphate pools are undetectable in vitamin B-12-deficient cells of Euglena gracillis, but appear rapidly after the replenishment with the vitamin. They reach a maximum size that is about 6 times that of normal exponentially growing cells, but decrease to almost zero as the cells divide. The pools expand again during the post-replenishment shortened cell cycle. However, the expansion takes place during rather than before the resumption of DNA synthesis. The maximum sizes reached are still larger than in normal cells. By using the protein-synthesis inhibitor cycloheximide and determining the pool size, we found that vitamin-deficient cells apparently accumulate a large amount of ribonucleoside triphosphate reductase apoenzyme, which lacks the vitamin B12 coenzyme. We showed that the production of the deoxyribonucleoside triphosphates is not closely coupled to DNA synthesis under our experimental conditions, and that the concentration of the deoxyribonucleoside triphosphate pools per unit of DNA synthesized is almost constant for all stages of growth examined.


Studies on cathepsin B in human articular cartilage.

The thiol proteinase cathepsin B (EC 3.4.22.1), previously called cathepsin B1, was assayed in human articular cartilage by its hydrolysis of the synthetic substrate alpha-N-benzoyl-DL-arginine 2-naphthylamide. The enzyme was activated by cysteine and EDTA and completely inhibited by iodoacetamide and HgCl2. It was also partially inhibited by whole human serum. Human osteoarthrotic cartilage had increased activity when compared with normal cartilage. Cathepsin B activity of normal cartilage was age-related, being high in juveniles and declining to low values in adult and elderly individuals. Cathepsin D and cathepsin B both exhibited a zonal variation through the cartilage depth; the surface cells appeared to contain more activity than those close to the subchondral bone.


The fate of bilirubin-IXalpha glucuronide in cholestasis and during storage in vitro. Intramolecular rearrangement to positional isomers of glucuronic acid.

1. In aqueous solution above pH7 bilirubin-IXalpha 1-O-acylglucuronide rapidly isomerizes to the non-C-1 glucuronides by sequential migration of the bilirubin acyl group from position 1 to positions 2, 3 and 4 of the sugar moiety. The transformations are enhanced by increasing the pH. Compared with the rates at 37 degrees C the transformations are rather slow at 0 degrees C. Virtually complete inhibition is observed at values below pH6. The isomerization at 25 degrees C and pH 7.4 is not affected by the presence in the solutions of a molar excess of human serum albumin. 2. Isomerization in bile kept at 37 degrees C at pH7.7-7.8 is probably non-enzymic, as the rates of change are similar to those observed under comparable conditions for aqueous solutions of glucuronides of bilirubin-1Xalpha and of azodipyrrole. 3. Analysis without delay of normal biles of man and rats collected at 0 degrees C over a maximum period of 10 min shows that the bilirubin-IXalpha mono- and di-glucuronides consist exclusively of the 1-O-acyl isomers. 4. The mixtures of the four positional isomers of bilirubin-IXalpha glucuronide found in freshly collected biles of man and rats with cholestasis probably originate from initially synthesized 1-O-acylglucuronide by the same mechanism of sequential migration as has been observed in aqueous solutions of conjugated bilirubin-IXalpha.


Troponin I from human skeletal and cardiac muscles.

1. Myofibrils from human skeletal muscle contained regulatory proteins exhibiting similar electrophoretic behaviour to those present in rabbit skeletal muscle. 2. All human skeletal muscles examined contained two forms of troponin I corresponding to the forms already characterized in fast and slow rabbit muscle. 3. The ratios of the amounts of the two forms of troponin I in different human skeletal muscles were not identical with the ratios for the type 1 to type 2 fibres published in the literature. The ratios could, however, be arranged in the same rank order. 4. Primate heart contained a single form of troponin I different in molecular weight and amino acid composition from the skeletal forms. 5. A monospecific antiserum to human cardiac troponin I was prepared in the sheep and shown not to react with the fast or slow skeletal-muscle forms of troponin I from human and other species. 6. The anti-(human cardiac-muscle troponin I) reacted with the cardiac troponin I from the human, baboon, rabbit and rhesus monkey. Positive reactions were also obtained with urea extracts of whole cardiac tissue.


The cellulase of Trichoderma koningii. Purification and properties of some endoglucanase components with special reference to their action on cellulose when acting alone and in synergism with the cellobiohydrolase.

1. Four principal endoglucanase components of Trichoderma koningii cellulase were separated and purified by gel filtration on Sephadex G-75, ion-exchange chromatography on DEAE- and sulphoethyl-Sephadex and isoelectric focusing. 2. All four endoglucanases hydrolysed CM-cellulose, H3PO4-swollen cellulose, cellotetraose and cellopentaose, but differed in the rate and mode of attack. 3. Attack on cotton fibre by the endoglucanases was minimal, but resulted in changes that were manifested by an increased capacity for the uptake of alkali, and a decrease in tensile strength. 4. All four endoglucanases acted synergistically with the exoglucanase [cellobiohydrolase; Wood & McCrae (1972) Biochem. J. 128, 1183-1192] of T. koningii during the early stages of the breakdown of cotton fibre, but only two could produce extensive solubilization of cotton cellulose when acting in admixture with the exoglucanase component. 5. The mode of action of the enzymes is discussed in relation to these synergistic effects. It is suggested that the results are compatible with the interpretation that the 'crystalline' areas of cotton cellulose are hydrolysed only by those endoglucanases capable of forming of forming an enzyme-enzyme complex with the cellobiohydrolase on the surface of the cellulose chains.


The purification and properties of the second component of human complement.

The second component of human complement (C2) was purified by a combination of euglobulin precipitation, ion-exchange chromatography, (NH4)2SO4 precipitation and affinity chromatography. The final product was homogeneous by the criterion of polyacrylamide-gel electrophoresis and represents a purification of about 4000-fold from serum with 15-20% yield. Component C2 comprises a single carbohydrate-containing polypeptide chain, with an apparent mol.wt. of 102000; alanine is the N-terminal amino acid. The molecule is rapidly cleaved by activated subcomponent C1s with the loss of haemolytic activity to yield two fragments with apparent mol.wts. of 74000 and 34000. These fragments are not linked by disulphide bonds and can be easily separated. A second protein isolated during the purification of component C2 was identified by its haemolytic and antigenic properties as complement Factor B, the protein serving an analogous function to component C2 in the alternative pathway. The protein, which is also a single carbohydrate-containing polypeptide chain, has an apparent mol.wt. of 95000 and threonine as N-terminal amino acid. The amino acid analyses of component C2 and Factor B are compared.


Comparison of the presence of immune complexes in Felty's syndrome and rheumatoid arthritis.

Evidence has been found to document the presence of circulating immune complexes in all patients with Felty's syndrome. The sera of all 12 patients studied showed intermediate complexes by analytical ultracentrifugation. The sera of 9 of 12 patients (75%) showed precipitin lines upon immunodiffusion against IgM rheumatoid factor. Both findings were statistically increased above those in a matched group of patients with classic rheumatoid arthritis. The presence of circulating immune complexes in the sera of the Felty patients was consistent with the observation that large inclusions containing IgG, IgM, and complement were phagocytized by normal polymorphonuclear cells when incubated with sera of Felty patients. Normal polymorphonuclear cells phagocytosed inclusions from 77% of Felty sera, compared with 27% classic rheumatoid arthritis sera. It is suggested that the uptake of immune complexes by polymorphonuclear cells plays a role in the neutropenia of Felty's syndrome.


Synthesis by an established lymphocyte cell line from a rheumatoid synovium.

Lymphocytes derived from the synovium of a patient with rheumatoid arthritis were shown to synthesize predominantly IgG as measured by an equilibrium binding assay. Chromatographic separation of the radiolabeled lymphocyte culture supernatant revealed rheumatoid factor activity associated with IgG and IgM. In addition, immunofluorescence studies on fixed lymphocytes demonstrated that the majority of cells stained positive for both IgG and rheumatoid factor.


Purification and properties of the phenprocoumon-induced decarboxyfactor X from bovine plasma. A comparison to normal factor X.

1. By a procedure involving adsorption to barium sulfate, chromatography on DEAE-Sephadex and QAE-Sephadex and preparative polyacrylamide gel electrophoresis, decarboxyfactor X was purified from plasma of phenprocoumon-treated cows. No contaminants could be detected in the final preparation by polyacrylamide gel electrophoresis and zone-electrophoresis. 2. The molecular weight of decarboxyfactor X, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis is approximately 55 000, which is equal to that of factor X. The protein consists of two polypeptide chains with molecular weights of 44 000 and 17 000. 3. Decarboxyfactor X has antigenic determinants in common with normal factor X. 4. The amino acid composition and aminoterminal amino acids of normal factor X and decarboxyfactor X are identical. 5. Less than one residue of gamma-carboxyglutamate could be detected per mole of decarboxyfactor X. 6. In the absence of Ca2+, normal factor X has a slightly higher electrophoretic mobility than decarboxyfactor X. In the presence of Ca2+ the mobility of factor X decreases considerably while the mobility of decarboxyfactor X remains unaltered.


Activation of decarboxyfactor X by a protein from Russell's viper venom. Purification and partial characterization of activated decarboxyfactor X.

1. Incubation of decarboxyfactor X with the factor X-activating enzyme from Russell's Viper venom revealed the generation of amidase activity towards Bz-Ile-Glu-Gly-Arg-pNA, but not of activity in blood coagulation. 2. The rate of activation of both factor X and decarboxyfactor X depends on the ability of the zymogens to bind Ca2+. The relationship between Ca2+ concentration and velocity of the activation reaction is sigmoid in the case of factor X, but hyperbolic with decarboxyfactor X. 3. Activated decarboxyfactor X was purified by powder column electrophoresis. 4. Identical changes of primary structure accompanied the activation of factor X and decarboxyfactor X. Identical molecular weight and common antigenic determinants were found in factor Xa and decarboxyfactor Xa. The amino acid composition was identical except for 12 glutamic acid residues in decarboxyfactor Xa and gamma-carboxyglutamic acid residues in factor Xa. 5. Unlike factor X, activated factor X has a very low electrophoretic mobility in the presence of Ca2+ at pH 8.6. This is probably due to self association of factor Xa under the influence of Ca2+. The electrophoretic mobility of activated decarboxyfactor X is only slightly decreased compared to decarboxyfactor X in the presence of Ca2+.


Purification and characterization of nerve growth factor from the venom of Bungarus multicinctus.

The nerve growth factor from Bungarus multicinctus venom was purified by means of successive chromatography on Sephadex G-50, carboxymethyl-cellulose, carboxymethyl-Sephadex C-50 and Sephadex G-50. The purified nerve growth factor was homogeneous by polyacrylamide disc gel electrophoresis. The molecular weight was estimated to be about 21 000 by gel filtration. Compared with the nerve growth factors from the venoms of other Elapidae, namely Naja naja and Naja naja atra, this protein showed high isoelectric point of approximately pH 10. A characteristic of the nerve growth factor of B. multicinctus venom is that the protein consisted of two subunits of equal molecular weight which are linked covalently to each other by a disulfide bridge. The purified B. multicintus nerve growth factor elicited its maximal neurite outgrowth from embryonic dorsal root ganglia at the concentration interval from 30 to 100 ng/ml.


Binary and ternary complexes of dihydrofolate reductase with substrates, coenzymes and inhibitors. Circular dichroic and magnetic circular dichroic studies.

Interaction of several representative folate, quinazoline and pyridine nucleotide derivatives with dihydrofolate reductase from amethopterin-resistant Lactobacillus casei induces dramatic changes in its circular dichroic spectral properties. The binding of dihydrofolate induces a large extrinsic Cotton effect at 295 nm ([theta] = 113 800 deg . cm2 . dm-1). The generation of this band by dihydrofolate is strictly dependent on complex formation with a single substrate binding site and a KD = 7 . 10(-6) M. The other binary complexes examined include the enzyme . NADPH, enzyme . amethopterin, enzyme . folate, and enzyme . methasquin. All such complexes differ in spectral detail, the negative ellipticity at 330 nm being characteristic of the "folate site" complexes. The circular dichroic spectrum of the ternary complex of reductase . NADPH . methotrexate shows a positive symmetrical band centered at 360 nm ([theta] - 32 000 deg . cm2 . dm-1). Since both of the corresponding binary complexes exhibit negative bands in this region, this induced band represents a unique molecular property of the ternary complex. Chemical modification of a single tryptophan residue of the enzyme, as determined from magnetic circular dichroism spectra, results in a complete loss in the ability to bind either dihydrofolate or NADPH.


The chemical modification of tryptophan residues of alpha-mannosidase from Phaseolus vulgaris.

Reaction of alpha-mannosidase (alpha-D-mannoside mannohydrolase, EC 3.2.1.24) from Phaseolus vulgaris with N-bromosuccinimide or 2-hydroxy-5-nitrobenzyl bromide- resulted in loss of enzyme activity. Spectral absorption and fluorescence studies, as well as amino acid analysis, suggested that only tryptophan residues had been modified. No change in conformation could be detected by density gradient ultracentrifugation or circular dichroism of alpha-mannosidase modified by N-bromosuccinimide to virtually zero enzyme activity. The inhibition was partly offset by the substrate analogue alpha-methyl-D-mannoside and the competitive inhibitor mannono-1,4-lactone. Concomitantly, two tryptophan residues fewer were oxidized per molecule. After modification V was reduced, while Km seemed unchanged. Further, there was found evidence for the enzyme having a secondary structure dominated by beta-pleated sheets.


Stabilization of casein mRNA by prolactin and glucocorticoids.

Prolactin injected into pseudopregnant rabbits led to a parallel enhancement of casein synthesis and casein mRNA concentration. When this stimulation was followed by a withdrawal of prolactin obtained by injections of bromocriptine, the rate of casein synthesis progressively diminished. In the presence of endogenous prolactin after the initial stimulation, the decline of casein synthesis was delayed. Hydrocortisone acetate injected with bromocriptine after the initial stimulation by prolactin was able to maintain a high rate of casein synthesis. Measurements of casein mRNA concentration by hybridization with casein cDNA indicated that in all cases the amount of casein mRNA was correlated with the magnitude of casein synthesis. This suggests that the lactogenic hormones, prolactin and glucocorticoids, which were previously demonstrated to be responsible for the enhancement of casein mRNA concentration are involved in their stabilization.


[A new protein from human plasma lipoproteins: isolation and partial characterization of apolipoprotein G].

A new apolipoprotein has been identified in VHDL1 and in HDL. This protein is immunologically distinct from already isolated apoproteins. It was isolated by column chromatography on hydroxylapatite. In polyacrylamide gel electrophoresis, its mobility is very close to that of apo D. The amino acid composition differs from those of the well characterized polypeptides of the human plasma lipoproteins. It contains glucosamine. The apparent molecular weight is 72 000 +/- 2 000 in the presence and absence of reducing agent. According to the ABCDEF nomenclature, this protein can be named apolipoprotein G (apo G). It is present in a lipoprotein distinct from the lipoproteins A and D among the VHDL1 : this new lipoprotein can be named lipoprotein G (LPG).


[Diphenoloxydase (DPOx) in red cell membranes].

Two diphenoloxydases (DPOx) with similar molecular weights (158 000) are found in normal human red cell hemolysates. One of these enzymes appears to be tightly bound to the membranes while the other is not, or at most only weakly bound.


[Effect of a magnetic field on the rate of H202 breakdown by catalase and by an Fe3+--EDTA complex].

The acceleration of H202 decomposition induced by catalase and the dimer complex [Fe3+(EDTA)]2 has been observed in a constant magnetic field. The effect increases with the field increasing up to 8000 Oe, reaching 20 +/- 5% and 24 +/- 5% for catalase and [Fe3+(EDTA)]2 respectively. The results are discussed within the hypothesis of a one-electron reaction mechanism using the models developed specially to explain the magnetic effects in radical reactions. It is supposed that the stage which is affected by the magnetic field is the electron transfer coupled with Fe3+O./2 paramagnetic species. The interpretation proposed does not exclude an alternative possibility of the magnetic effects during the electron transfer to two iron atoms by a two-electron (synchronous) mechanism.


[The role of teichoic acids in the regulation of biochemical processes in microorganisms].

The role of cell wall teichoic and lipoteichoic acids in the regulation of activity of some autolytic enzymes and in ion exchange, the role of lipoteichoic acids in biosynthesis of cell wall teichoic acids and some other functional features of these compounds in the bacterial cell are discussed.


Structure-activity studies of aldose reductase inhibitors containing the 4-oxo-4h-chromen ring system.

Various 4-oxo-4H-chromen-2-carboxylic acids and their derivatives were screened for aldose reductase inhibitory activity. Their inhibitory response along with that of several flavonoids has been correlated with simple Hückel molecular orbital calculations. From these results a possible mode of action is postulated.


Kinetics of enzymatic hydrolysis of cellulose: analytical description of a mechanistic model.

A generalized mechanistic model for the enzymatic hydrolysis of cellulose is developed and expressed mathematically. The model is based on Michaelis--Menten-type kinetics for concurrent random and endwise attack of the substrate involving end-product inhibitions and three types of enzymes: an endo-beta-1,4-glucanase, an exo-beta-1,4-glucanase, and beta-glucosidase. Basic parameters of the model which can explain synergistic and other effects observed experimentally are quantified and discussed. It is shown that cellulose degradation kinetics are expected to be strongly affected by the ratio of endo- to exocellulases in the reaction mixture as indicated by previous experimental data, and the substrate degree of polymerization, a factor not fully appreciated in previous studies, which appear to be overridingly important in many practical cases.


The effects of a polychlorinated biphenyl)Aroclor 1254) on the white pelican: ultrastructure of hepatocytes.

White pelicans were each given 100 mg of polychlorinated biphenyls (Aroclor 1254) per day for 10 weeks. After the treatment period, the bird's food supply was decreased 50% for 2 weeks. Measurements taken from transmission electron micrographs of the pelican's liver tissue revealed hepatocytes of the treated birds averaged 22% larger in cross-section area than those of controls. The number of vacuoles (lysosomes, microbodies) per hepatocyte was 22% greater. The number of mitochondria per hepatocyte was 25% greater while 25% fewer cristae per mitochondrion were present in treated as compared to control birds.


The effect of polychlorobiphenyls (Aroclor 1242) on bicarbonate-C14 uptake by Euglena gracilis.

Aroclor 1242 is inhibitory to long-term batch growth of Euglena gracilis at 10 ppm. Exposure to PCB's, subsequent to an initial drop of 50% in the first 30 minutes, does not appear to inhibit photosynthesis on a per cell basis over the time span of four hours. Consequently our experimental findings suggest that inhibition of population growth does not lie directly in the photosynthetic pathway despite the initial inhibition by Aroclor 1242 on Euglena gracilis in "light". "Dark" absorption of bicarbonate appears to decrease with increased PCB exposure. This decrease may be causally related to the population growth inhibition observed.


Radiotherapy in the treatment of Hodgkin's disease.

Eighty-seven untreated patients with localised Hodgkin's disease seen from 1969 to 1975 were treated by megavoltage radiotherapy. All were followed for at least 33 months. Thirty-three patients were staged clinically and 54 underwent more extensive investigation by lapaortomy and splenectomy. The projected five-year disease-free survival figures for patients staged surgically were 100% for the 17 with stage IA disease, 70% for the 19 with stage IIA disease, and 73% for the 15 with stage IIIA disease. These results were consistently better than those obtained in clinically staged patients. Five patients died, one of them without evidence of Hodgkin's disease. As irradiation seems to produce excellent disease-free survival in most patients who are staged accurately at diagnosis, caution should be exercised in the routine use of adjuvant chemotherapy until the full risks of such treatment are clear. Combined modality therapy may be appropriate for patients with unfavourable features at presentation.


Periurethral aerobic microflora of pregnant and non-pregnant women.

Seventy-two pregnant and 88 non-pregnant women were examined to see whether the periurethral region had been colonised with group B streptococci (Streptococcus agalactiae), enterococci, and Gram-negative rods belonging to the Enterobacteriaeceae. A semi-quantitative method was used for periurethral sampling, and paired urethral swabs were also collected to compare the isolation rates of group B streptococci from the two sites and with the two sampling methods. A higher isolation rate was found with periurethral sampling. Most specimens showed no or scanty growth of Gram-negative rods. Pregnancy was often associated with heavy growth of enterococci. Sampling performed during menstruation and while oral contraceptives were being used produced high isolation rates of group B streptococci. These results seem to suggest that the periurethral area might protect against genital colonisation with group B streptococci as it does against urinary tract infection and that hormonal factors influence the carriage of these organisms.


Prevention of secondary cases of meningococcal disease in household contacts by vaccination.

Household contacts of patients with group A meningococcal infection were vaccinated with either meningococcal vaccine or tetanus toxoid. Five of the 523 subjects who received tetanus toxoid developed meningococcal meningitis and another four probably had meningococcal disease. Only one possible case of meningococcal infection occurred among 520 contacts vaccinated with meningococcal vaccine. Vaccination had no effect on nasopharyngeal carriage of meningococci. Vaccination of household contacts of patients with group A meningococcal infections is an effective way of using limited supplies of meningococcal vaccine, though its value would be limited in an epidemic. Secondary cases of meningococcal infection often occur within a few days of the index case, and, although vaccine alone seemed to provide adequate prophylaxis in these Nigerian subjects, additional chemoprophylaxis may be needed to cover this critical period.


Saccades induced by stimulation of the frontal eye fields: interaction with voluntary and reflexive eye movements.

Saccadic eye movements were evoked in alert monkeys by electrical stimulation of the frontal eye fields (FEF). The interaction of FEF movements with voluntary saccades, pursuit, and vestibular nystagmus (VN) was examined. An FEF saccade in one direction could be evoked as little as 10 msec after the onset of a natural rapid movement (saccade, fast phase of VN) in the opposite direction. Thus, refractory periods between similar eye movements are directionally specific. The frequency of evoked FEF saccades following natural slow movements (pursuit, slow phase of VN) in either direction was reduced as though saccade thresholds were elevated. The FEF appears capable of modulating any ongoing eye movement.


The effects of bilateral frontal eye-field, posterior parietal or superior collicular lesions on visual search in the rhesus monkeys.

Superior collicular, frontal eye-field and posterior parietal lesions increased the time taken to find a circular target among other geometrical shapes. The collicular monkeys were considerably more impaired than the other groups, suggesting that the superior colliculus is the main neural structure underlying this highly practised visual search task. Failure to respond in the 5 sec allowed on each trail increased for all groups, and the collicular and frontal groups showed a small increase in false positive errors. None of the groups increased search time for a near-threshold target on a homogenous background. [corrected] Analysis of the latencies of individual trials suggest that scanpaths over the display were still systematic after the lesions. This suggests that the search was being slowed down either by the need to make more correction saccades to give accurate fixations of the stimuli being discriminated, or by an increase in latencies specificially for those saccades that shift the gaze between stimuli.


Operant conditioning of tonic firing patterns from precentral neurons in monkey neocortex.

This report presents a single neurons operant conditioning paradigm which allows the quantification of operant control between neurons and monkeys. The operant task is for the monkey to change firing patterns of the neuron from phasic to tonic. In 60 neurons conditioned after the protocol had been standardized the following results were obtained. (1) For a fixed interspike interval target range, the time off target may be considered error, and this may be used as a parameter with which to judge operant neuronal control. (2) The degree to which the monkey could control a neuron was not correlative with the neuron's initial firing rate, firing rate variance, or pattern. (3) Neurons coactivated by distal arm muscle groups were, as a group, more highly controlled in comparison to neurons coactivated by proximal muscle groups. (4) Pyramidal tract neurons, as a group, appear more accurately controlled than non-pyramidal tract neurons. (5) The role of proprioception in response acquisition is discussed.


Comparison of forelimb and hindlimb motor deficits following dorsal column section in monkeys.

Macaca speciosa monkeys were trained to acquire food reinforcement with motor responses that were defined by 4 different tasks. The effects of dorsal column lesions on the speed of these responses were compared for the forelimbs vs. the hindlimbs. Enduring impairments were not seen for any limb when the animal was required to accurately project a limb to different points in space, even with exclusion of visual guidance and with random variation of the start and stop points for each movement. Similarly, when the task required that the animals emit rapid response sequences, the forelimbs were impaired temporarily, but no long-term deficits were seen for the forelimbs or the hindlimbs. Although these impositions of spatial and temporal demands did not reveal striking disruptions of whole-limb movements, hindlimb grasp responses were shown to be impaired over long periods of postoperative testing. This corroborates previous findings for the forelimb and indicates that facility of distal extremity movement depends crucially on dorsal column-lemniscal input.


Categorization and retrieval after selective inferotemporal lesions in monkeys.

Two variations of a colour matching to sample task, with and without delay, were used to explore the deficits produced by bilateral foveal prestriate, posterior inferotemporal or anterior inferotemporal cortex lesions in monkeys. In the first task, 7 colours served as samples, but matching had to be done from only 4 alternative stimuli (7s x 4a); this task taxes the animal's ability to categorize information. In the second variation of matching to sample only 4 colours served as samples, but matching had to be done from amongst 8 alternative stimuli (4s x 8a); here, ability to retrieve information is tested. Task comparisons showed that the posterior inferotemporal group performed poorly on the 7s x 4a problem, whilst the AIT group were worse on the 4s x 8a task. It is concluded that posterior inferotemporal lesions disrupt categorization, whereas anterior inferotemporal lesions affect the retrieval of information.


Use of pneumoencephalography to increase stereotaxic accuracy in rhesus monkeys.

A radiographic method for visualizing intracranial structures for stereotaxic surgery is described. The procedure circumvents the problems associated with the use of radiopaque dyes, and it does not require modification of standard stereotaxic equipment. Anatomical data derived from the use of the method are presented, as well as an example of its application for accurately placing cannulae in the midbrain.


Technique for casting chronic extradural and intracerebral cannula arrays.

A technique for fixing multiple brain cannulae in uniform arrays by embedding guide cannulae in molded plastic pedestals is described. Methods for producing corresponding threaded caps for protection of the outer ends of the cannulae, and procedures for reducing mortality and morbidity when large number of cannulae are implanted in brain tissue, are presented.


Influence of EGTA on an exercise-induced elevation in the colonic temperature of the rat.

The purpose of this study was to determine if an exercise-induced rise in body temperature could be affected by the chelation of Ca++ in the extracellular fluid surrounding the hypothalamus of the rat. Following the implantation of a guide tube above the hypothalamus, each animal was familiarized with exercising on a motor-driven treadmill. In random order, on separate days, a solution containing 3.6 mM EGTA, 26 mM Ca++ or an artificial cerebrospinal fluid (ACSF) solution was perfused through the guide tube while the animal was running. Colonic (Tc) and tail-skin (Tt) temperatures were monitored continuously. The perfusion of EGTA produced a significant increase in Tc when compared with the perfusion of the ACSF solution. The perfusion of excess Ca++ produced a significant decrease in Tc. These results suggest that Ca++ may play an important role in the mediation of heat dissipation during exercise.


The use of case history studies to differentiate potentially infected from potentially noninfected herds with reactors to Brucella abortus antigens.

Data on farm characteristics and the results of the first herd test for brucellosis were collected for 74 reactor and 74 negative herds in Wellington County, Ontario. Each reactor herd was classified as either probably infected or probably not infected using the occurrence of abortions prior to the first herd test or during the testing period, the total number of cattle removed and/or the spread of reactors within the herd as criteria of infection. Statistical techniques were used to select variables which were good "discriminators" between probably infected and noninfected herds. In general, reactor herds were primarily dairy herds and were somewhat larger than negative herds. The presence of only single suspicious reactors on the first test appeared to be a good predictor of lack of infection with Brucella abortus. Among the 37 farms in this category the single reactor was removed from only eight farms and no evidence o fthe spread of infection was observed. The presence of one or more positive reactors on the first herd test appeared to be a good predictor of the presence of infection. Of the 24 farms in this category, evidence of the spread of infection was present in ten farms and seven of these ten farms were eventually depopulated. The brucella milk ring test appeared to be the most effective means of identifying infected herds under the conditions present in Wellington County.


The effects of Brucella strain 19 vaccination on titers in the tube agglutination test: a case control study.

The degree of agglutination in the tube agglutination test of the sera of 373 "reactor" cattle was compared to that of 800 "non-reactor" cattle from multiple reactor herds in southwestern Ontario. Vaccinated cattle had lower titers than did nonvaccinated cattle. Thus, vaccinated cattle were less likely to be classified as positive than nonvaccinated cattle irrespective of the method of interpreting the results of the tube test. When the full allowance for vaccination was used, vaccinated cattle were less likely to be classified as suspicious than nonvaccinated cattle. The extent of herd vaccination did not appear to markedly influence the effects of vaccination or the prevalence of reactors. Thus, it appeared that vaccination with strain 19 produced a small protective effect (inferred from the lower titers in vaccinated animals), but that the magnitude of this effect was markedly dependent on the appropriateness of the vaccine allowance for classifying cattle. Further work is needed on this aspect of vaccination with strain 19.


Studies on circulating gonococcal antibodies and antigens.

One hundred human sera obtained from acute gonococcal disease and 100 sera from nongonococcal diseases or healthy persons were concentrated four times and examined for the presence of circulating gonococcal antigens and antibodies by means of a counterimmunoelectrophoresis (CIE) and delayed hypersensitivity assay. Antibodies reacting with cytoplasmic gonococcal antigens were detected by CIE in 92% of sera received from patients suffering from acute gonococcal disease. Gonococcal antigens were found in the concentrated sera of 82.3% of patients on the basis of dermal reactions observed upon injections of these sera into the skin of rabbits sensitized with disrupted gonococci; 51.8% of the patients' sera gave delayed hypersensitivity reactions in rabbits sensitized with cytoplasmic antigens of N. gonorrhoeae. Control sera from healthy people and those with non-gonococcal diseases did not react with any of the preparations tested.


The lipopolysaccharide (R type) as a common antigen of Neisseria gonorrhoeae. I. Immunizing properties.

The ability of R-type lipopolysaccharide (LPS), isolated from Neisseria gonorrhoeae colony type 4, to protect against infection with N. gonorrhoea colony type 1 (T1 isolates) in the mouse and chicken embryo was investigated. C57 black mice were immunized intraperitoneally with 50 microgram of LPS, and challenged intracerebrally with 10-20 LD50's of N. gonorrhoeae colony type 1. Immunized mice were significantly protected (P less than 0.01 to less than 0.05) against challenge with different T1 isolates of N. gonorrhoeae when compared with non-immunized mice. Mice, injected with succinylated or alkali-treated LPS were not protected against gonococcal challenges. In a second animal model, leghorn hens were immunized intravenously with three injections of 500 microgram of LPS followed by a booster of 2.5 mg 2 weeks later. Embryonated eggs obtained from immunized hens were protected against challenge with 5 x 10(3) - 1 x 10(4) LD50's of three different T1 isolates. When hens were injected with the chemically modified LPS, the embryos were not resistant to gonococcal challenge. The results of this study demonstrate the ability of R-type gonococcal LPS to provide protection against different T1 isolates of N. gonorrhoeae.


The lipopolysaccharide (R type) as a common antigen of Neisseria gonorrhoeae. II. Use of hen antiserum to gonococcal lipopolysaccharide in a rapid slide test for the identification of N. gonorrhoeae from primary isolates and secondary cultures.

An antiserum has been prepared in hens to R-type gonococcal lipopolysaccharide (LPS) and used in a simple slide-agglutination test for the identification of Neisseria gonorrhoeae. Anti-LPS serum agglutinated gonococcal cells representative of the four colony types of N. gonorrhoeae. Absorption of the antiserum with LPS removed the agglutinating activity. Secondary cultures (1120) were tested without observation of the colony type and all were agglutinated. No agglutination occurred with strains of Neisseria meningitidis, Neisseria lactamica, non-pathogenic Neisseria. Pseudomonas aeruginosa, Branhamella catarrhalis, or with species of lactobacilli and Acinetobacter. Cross-reactivity of the antiserum occurred with some streptococci. The anti-LPS serum was used to identify N. gonorrhoeae in primary isolates from the cervix, urethra, and pharynx. Of 251 gonococcal isolates tested, 249 were agglutinated by the antiserum, while all of the corresponding second cultures were agglutinated. The antiserum did not agglutinate N. meningitidis found in primary isolates from pharyngeal specimens. Anti-LPS hen serum should be useful for the rapid identification of N. gonorrhoeae in primary isolates or secondary cultures.


[Diminution of the antibacterial activity of antibiotics in cultures and in experimental mixed infections].

We have studied interactions between Staphylococcus aureus and Pseudomonas aeruginosa in the absence and the presence of four different antimicrobials in mixed cultures and experimental infections. These two bacterial species, in addition to having different properties, are known to be opportunistic pathogens often present in human microflora. Two main aspects have been investigated and they are related to modifications in two species affecting their equilibrium in the mixed bacterial population and also their pathogenicity markers. Our results indicate that individual growth of S. aureus and P. aeruginosa is not modified in vitro in mixed cultures in the absence of antimicrobials; in vivo, in mouse peritoneal cavity, there is a synergism favorable to S. aureus. In the presence of rifamycin SV and three cell wall inhibitors, pencillin G,D-cycloserine, and vancomycin, we have observed that P. aeruginosa protected S. aureus against the inhibitory effect of these antimicrobials in vitro and in vivo. Such results were obtained in different conditions of culture, stationary, shaken, and in special apparatuses, an "Ecologen" and a "Chemostat." When any one of the antimicrobials was allowed to be in contact  for 6 to 8 h with P. aeruginosa cells in a culture, we observed a decrease in their inhibitory effects against S. aureus. These results were supported by microscopical observation. It seems that the inhibitory effects of the antimicrobials have hindered the formation of toxic products of S. aureus, e.g., alpha toxin, and that it was not restored in the presence of P. aeruginosa. Conversely, P. aeruginosa remained apparently unchanged through all these experiments. Our observations may imply that the inhibitory effect of an antimicrobial towards a bacterial species may be significantly decreased in the presence of another species, sometimes present in human microflora.


Neisseria gonorrhoeae: evaluation of some methods used for carbohydrate utilization.

A total of 989 Neisseria gonorrhoeae cultures were examined for acid production from dextrose in seven different carbohydrate media. Four of these media were found to be quite accurate for this purpose. Some pitfalls in the preparation and usage of cystine trypticase agar carbohydrate media are discussed. The term 'fermentation' of carbohydrates by Neisseria species is inaccurate and responsible for some N. gonorrhoeae false-dextrose-negative results.


Health--indicator bacteria in water--surface microlayers.

The distribuution patterns of a variety of bacteria in the surface microlayer, 20 cm and 100 cm below the surface of rivers, open lake waters, and off docks are presented. Implications of this three-season study are that conventional sampling techniques underestimate the true microbiol densities.


Immunological relationships of some non-pathogenic Neisseria.

A collection of nine stock strains of non-pathogenic Neisseria species and one strain of Branhamella catarrhalis has been examined serologically by a simple immunodiffusion technique utilising cultures growing on solid medium.


Protein-lipopolysaccharide interactions. 1. The reaction of lysozyme with Pseudomonas aeruginosa LPS.

Lysozyme (EC 3.2.1.17) complexes with extracted Pseudomonas aeruginosa LPS in two distinct stages. The initial stage does not produce turbidity detectable by nephelometry (measured as nephelos units (N) per time) but does permit low-speed sedimentation of the lysozyme-lipopolysaccharide (LPS) complex. This association is 100% disrupted by the action of 0.1 M Mg2+. Monovalent cations at equal ionic strength to the Mg2+ concentration used for these studies failed to alter significantly the lysozyme-LPS complex, indicating that the role of Mg2+ was not strictly an ionic one. The study of lysozyme-LPS complexes may provide a model system for investigating in vivo protein-LPS interactions.


Colonization resistance against potentially pathogenic bacteria in hexaflora-associated gnotobiotic mice.

A study of colonization resistance against potentially pathogenic bacteria (Escherichia coli and Pseudomonas aeruginosa) was conducted in hexaflora-associated gnotobiotic mice. Groups of germfree AKR mice were swabbed with five bacterial and a single gastrointestinal yeast species: Streptococcus faecalis. Lactobacillus brevis. Staphylococcus epidermidis, Enterobacter aerogenes, Bacteroides fragilis var. vulgatus, and Torulopsis sp. All species became established in the gut in 8 weeks. Later these associated mice were divided and challenged by four graded doses of E. coli or P. aeruginosa. The presence of challenge organism was monitored specifically in the freshly voided fecal specimens of the challenged mice. Escherichia coli colonized the gut of each mouse at each level up to 60 days post challenge. Pseudomonas aeruginosa was completely eliminated from each mouse at each dose level after 30 days post challenge. Evidence suggests that all six species were sufficient to prevent the colonization of P. aeruginosa and not of E. coli in the gut of the gnotobiotic mice.


[Effect of surface-active agents on Mycobacterium tuberculosis and Mycobacterium bovis. Obtaining chromogenic mycobacteria].

In 18 strains of M. tuberculosis and M. bovis it has been possible to demonstrate the existence, in pure cultures, of elements able to grow in vitro in scotochromogenic mycobacteria. The experimental method used includes a dispersion, under shaking, of organisms in different surface-active agents. The chromogenic strains isolated during the experimentation are eugonic, and PAS- and INH-resistant. Their experimental virulence is different from that of tubercle bacillus. We discuss the signification of this dissociation phenomenon inductible by surface-active agents.


Genetic fate of DNA in a strain of Bacillus subtilis which is impaired in genetic transformation.

A mutation in Bacillus subtilis call recC4 which results in an impairment of genetic transformation was transferred to a new strain using the closely linked marker mit-2 (mitomycin C-resistance) for selection. This derived strain was in turn impaired in transformation but showed normal levels of sensitivity to ultraviolet irradiation and methyl methane sulfonate. The genetic and molecular fate of transforming DNA in the recC4 strain was studied. Normal amounts of DNA were taken up by the cells and this DNA or parts of it became associated with recipient DNA. Linkage between genes on donor and recipient molecules was, however, not established and transformants were not generated. The recC4 mutation therefore affects a step in the recombination pathway during transformation. Either the association between donor and recipient DNA molecules is abnormal or the cells are deficient in the further processing of the associated complex.


Biochemical identification of a tryptophan auxotroph in Rhodospirillum rubrum.

A tryptophan auxotroph of aerobically grown Rodospirillum rubrum was isolated after mutagenesis and replica plating. The mutant grows on tryptophan or indole, accumulates indole glycerol phosphate, lacks the capacity to convert indole glycerol phosphate to indole glycerol phosphate to indole, and finally is defective in photosynthetic growth. The strain is, therefore, analogous to a Trp A mutant which is defective in the alpha-subunit structural gene of tryptophan synthetase.


Ovarian carcinoma terminating in acute nonlymphocytic leukemia following alkylating agent therapy.

Rapidly fatal acute myelogenous leukemia (AML) occurred in a woman with advanced (Stage III) ovarian carcinoma who was treated with thiotepa for 30 months. This patient was 1 of 10 long term survivors and represented less than 2% of patients with advanced ovarian carcinoma with regional metastases who received long term chemotherapy during the period 1947-1975. Acute leukemia developed 44 months after initial diagnosis and was preceded by a 10 month period of pancytopenia following cessation of thiotepa. The leukemia did not respond to treatment and the patient expired 3 weeks after its onset. At autopsy, leukemic infiltration of organs was seen, but there was no evidence of carcinoma. A review of the literature suggests that the development of AML reported in ovarian cancer patients is related to alkylating agent therapy.


Dose response in radiotherapy for glottic carcinoma.

All consecutive patients with glottic squamous cell carcinoma treated primarily by radiotherapy from 1965 through 1974 have been analyzed to reveal response of the primary tumor in relation to the dose. This study appears to indicate presence of dose response for all stages of the primary lesions (T) of the glottic carcinoma except for the smallest ones namely TIS, T1a and T1b. The total number of advanced tumors is small and selected. But they tend to indicate presence of dose response. Out of 15 selected patients with T3 and T4 glottic tumors, 5 (33%) had attained local control by primary radiotherapy; two had distant metastasis at onset of therapy, while 5 (33%) needed salvage surgery for control of the residual or recurrent disease. The ultimate success rate is 66% (10 out of 15) for the patients with advanced tumors treated with a conservative approach. An ongoing study indicates similar, if not better preliminary results with high dose radiotherapy.


Radiotherapy of Kaposi's sarcoma.

Between 1958 and 1976, 34 patients with Kaposi's sarcoma were seen. Two patients were kidney transplant recipients. Co-existing malignancies were seen in 22% of patients. From 1958 to 1965, cutaneous lesions were treated solely with local radiotherapy techniques, single doses of 800 rads being found adequate to produce a complete response. In 1965, because of the multicentric occurrence of the disease and frequent recurrences after local radiotherapy techniques, extended field radiotherapy was begun. Ten of twelve patients thus treated responded completely. It is concluded that extended field radiotherapy using a single dose of 800 rads offers complete relief of symptoms and better control of the disease when compared to local radiotherapy. There was very little morbidity, with the extended field technique.


Radiation myelopathy of the thoracic spinal cord in long term survivors treated with radical radiotherapy using conventional fractionation.

Previous reports on radiation myelopathy of the thoracic spinal cord have attempted to predict tolerance doses as a function of overall treatment time and number of fractions by extrapolation of data from collected myelopathy cases having markedly heterogeneous treatment parameters. A review of long term survivors receiving radical radiotherapy with conventional fractionation for lung and esophageal cancer was undertaken to determine if the proposed tolerance levels were consistent with clinical results in a series of patients constituting the group at risk for developing myelopathy. Spinal cord doses were determined in 58 patients who survived greater than or equal to 18 months following radiotherapy. All patients were treated with megavoltage equipment at dose rates of 200 rads/day continuously (53 patients) or 250 rads/day using split course (5 patients). Tumor doses ranged from 4500 rads to 7000 rads. Seventeen patients (29%) received less than 5000 rads to the thoracic cord, 6 patients (10%) received 5000 rads, and 35 patients (61%) received greater than 5000 rads. There were two patients who developed myelopathy. Review of our data, the clinical data of others and recent experimental results suggest that the slope of tolerance curves for thoracic spinal cord commonly utilized to express time-dose-fractionation relationships may be inappropriate. It is suggested that cord tolerance to well fractionated radiation has been previously underestimated.


Leukemic reticuloendotheliosis: polyclonal surface immunoglobulin on "hairy" cells.

The cytochemistry, surface markers and functional properties of purified mononuclear cells obtained from the peripheral blood and spleen of a patient with leukemic reticuloendotheliosis were studied. Nonspecific esterase activity, a monocyte marker, was demonstrable in 83% of the peripheral blood mononuclear cells and 84% of the splenic mononuclear cells. Rosetting techniques failed to detect T or B lymphocyte surface markers on the majority of the cells. Direct immunofluorescence revealed capped, noncytophilic surface immunoglobulin on the cells with all immunoglobulin classes being detectable. Since noncapping conditions ahd been used during immunofluorescence staining, the observed caps were attributed to in vivo binding of autoantibodies to the "hairy" cells. This conclusion was supported by the demonstration of susceptibility of the "hairy" cells to lysis mediated by normal allogeneic lymphocytes. It is postulated that the "hairy" cells in this patient are leukemic monocytes which bear autoantibodies directed against leukemia associated antigens.


Pleomorphic reticulum cell sarcoma, monoclonal gammopathy and amyloidosis: an immunoperoxidase study.

Pleomorphic reticulum cell sarcoma, a histologic variant of the histiocytic lymphomas, presented as an abdominal mass in a 52-year-old woman. Extensive amyloid deposition was present within the tumor mass and an M-component (IgG Lambda) was identified in the serum. Direct immunoperoxidase staining of tissue sections demonstrated the same monoclonal immunoglobulin to be present within the neoplastic cells, presumptive evidence of their ability to both synthesize and secrete immunoglobulin. The presence of amyloid within this patient was probably the direct result of the tissue deposition of this monoclonal immunoglobulin and may be related to the amyloidogenic nature of Lambda light chains. Immunoglobulin production is a specific property of the B lymphocyte series. Transforming B lymphocytes and their differentiating progeny are engaged in active immunoglobulin synthesis and thus may be distinguished from morphologically identical but functionally distinct cells. The demonstration of cytoplasmic monoclonal immunoglobulin within these "malignant reticulum cells" strongly supports the assertion that at least some "histiocytic lymphomas" are neoplastic analogues of transformed B lymphocytes, and are not derived from phagocytic histiocytes, as previously believed.


An aggressive approach to marginally resectable lung cancer.

Between July 1968 and December 1974, 53 patients with lung cancer were planned for preoperative irradiation and surgery. All patients were considered clinically marginally resectable because of advanced local disease, 4 Stage II patients, with limited pulmonary reserve and 49 Stage III patients. Most patients received 3000 to 4000 rad followed in two weeks by thoracotomy. Forty-six patients were explored and 38 were resectable. Twelve patients are alive with a median follow-up of 48 months. The cumulative 5-year survival of all resectable patients is 27%. The survival of patients with marginally resectable lung cancer treated by accelerated radiotherapy followed by aggressive surgery approaches the survival experience of patients with primary resectable lung cancer and is superior to such patients treated with radiation therapy alone.


Anterior resection and primary anastomosis following high dose preoperative irradiation for adenocarcinoma of the recto-sigmoid.

Preoperative irradiation for adenocarcinoma of the rectum and sigmoid does not always limit the surgery to an abdominoperineal resection. From 1960 to 1976 anterior resection and primary anastomosis of the bowel has been performed in 13 patients with adenocarcinoma of the rectum and sigmoid whose tumor had been irradiated with 5000 rads with small pelvic fields. The inferior surgical resection line was within or very near the edge of the radiation field in 10 patients. In no instance was the superior resection line irradiated. Compared to a group of 79 patients treated with anterior resection only, the preoperatively irradiated patients had lower incidence of pelvic and anastomotic tumor recurrence, but a higher incidence of anastomotic leak and subsequent adhesions and intestinal obstruction. We stress that if irradiated rectum is considered for forming the anastomosis, a temporary "protective colostomy" should be strongly considered at the time of the surgery.


Cell and microsome mediated binding of 7,12-dimethylbenz(a)anthracene to DNA studied by fluorescence spectroscopy.

Fluorescence spectra of DNA isolated from hamster embryo cells incubated with 7,12-dimethylbenz(a)anthracene, or DNA modified in a microsomal system by reaction with this carcinogen or its 7-hydroxymethyl derivative, were compared to various model compounds. The spectra indicate that the DMBA derivative bound to DNA, in all 3 cases, has a 9,10-dimethylanthracene-like chromophore. They also provide the first evidence of the similarity in structure of the DNA-bound products between 7,12-dimethylbenz(a)anthracene and its 7-hydroxymethyl derivative. Our results are consistent with an activation mechanism that involves saturation of the 1,2,3,4-ring positions.


Production of pancreatic acinar cell carcinoma by combined administration of 4-hydroxyaminoquinoline 1-oxide and azaserine in partial pancreatectomized rats.

The effect of azaserine on the pancreatic tumorigenesis of 4-hydroxyaminoquinoline 1-oxide (4-HAQO) after partial pancreatectomy in rats was studied. Pancreatic acinar cell carcinomas were produced in 7 out of 10 rats (70%), which received 7 mg/kg body wt. 4-HAQO 3 days after partial pancreatectomy, followed by 10 weekly injections of 30 mg/kg body wt. azaserine. Partial pancreatectomy enhanced the carcinogenesis of 4-HAQO, which was further promoted by azaserine.


Biochemical activation of aryl hydrocarbon hydroxylase activity, cellular distribution of polynuclear hydrocarbon metabolites, and DNA damage by polynuclear hydrocarbon products in human cells in vitro.

Carcinogenic polynuclear hydrocarbons [7,12-dimethylbenzanthracene, 3-methylcholanthrene, and benzo(a)pyrene] were added to human skin fibroblast cell cultures. Only benzo(a)pyrene at 10 microgram/ml or above induced mixed-function hydroxylase activity, altered cell proliferation kinetics, and caused DNA damage as measured by altered grain count and bromodeoxyuridine incorporation. 3-Methylcholanthrene at concentrations as high as 15 microgram/ml was ineffective. 7,12-Dimethylbenzanthracene at 6 microgram/ml or above induced mixed-function oxygenase and stimulated DNA synthesis and cell proliferation, but at those concentrations little or no cytotoxicity or DNA damage was detected. The noncarcinogenic analogs 6,8,12-trimethylbenzanthracene, 5-fluorodimethylbenzanthracene, anthracene, and phenanthrene had no detectable effect on the human cells. It was concluded that benzo(a)pyrene can initiate all the biochemical events in human cells probably necessary to initiate transformation of human cells in vitro.


Actions of glycogen synthase and phosphorylase of rabbit-skeletal muscle on modified glycogens.

The high reactivities exhibited by rabbit-muscle synthase and phosphorylase for unmodified glycogen-acceptors decrease progressively, presumably because of a large increase in apparent Km as the glycogen molecule is converted into its component maltosaccharide chains by the debranching enzyme, isoamylase. Elongation of the outer chains of glycogen acceptor also results in decreased reactivities of the two transglucosylases and this is shown, for phosphorylase acting in the direction of glucan synthesis, to be caused by a decrease in the Vmax of the reaction. A partial restoration of the degradative reactivity of phosphorylase by a limited alpha-amylolysis of the long outer-chains of modified glycogen suggests a role of cytoplasmic alpha-amylase in mammalian glycogen metabolism.


Anterior pituitary response to thyrotrophin-releasing hormone during open-heart surgery.

The anterior pituitary response to thyrotrophin-releasing hormone has been studied in 20 patients submitted to elective open-heart surgical procedures, and in six control patients submitted to closed mitral valvotomy. Standard non-pulsatile normothermic perfusion was used in all the open-heart cases. 400 microgram thyrotropin-releasing hormone was administered by intravenous injection during bypass, at 30 min post-bypass, and at 60 min post-bypass. The same dose (400 microgram) was given during closed mitral valvotomy (Control Group). Thyrotrophin-releasing hormone administration during bypass failed to produce a normal response from the anterior pituitary, in contrast to the normal response pattern seen in the control group (P less than 0.01). Thyrotrophin-releasing hormone given after the period of bypass produced responses within the normal range in the majority of patients. These results suggest that anterior pituitary hypofunction may exist during the period of extracorporeal circulation using non-pulsatile perfusion and that recovery of pituitary function is evident within the first hour post-extracorporeal circulation.


Cytochalasin B and water transport. A scanning electron microscope study of the toad urinary bladder.

A morpho-functional study of the effects of cytochalasin B (CB) on Na and water transport was made in amphibian epithelia. The functional studies confirmed the dissociation of the natriferic and hydrosmotic effects of vasopressin in toad urinary bladders exposed to CB and showed in addition that the block of the hydrosmotic effect was reversible and could still be induced in epithelia maximally stimulated with the hormone. Scanning electron microscopy revealed that CB, per se, did not alter the apical surface of the bladders. An almost total loss of microvilli of granular cells was seen, however, if CB was associated with vasopressin and an osmotic gradient. The results suggest two points: a) the block of the hydrosmotic flow induced by CB is due to factors beyond the apical membrane; b) microfilaments may be important mechanochemical transducers in the chain of events leading to the hydrosmotic effect of vasopressin.


Formation of 7,12-dimethylbenz[alpha] anthracene-DNA adducts in 7,8-benzoflavone-treated hamster embryo cells.

Pretreatment of secondary cultures of Syrian hamster embryo cells with 7,8-benzoflavone (7,8-BF) inhibited both the metabolism of 7,12-dimethylbenz[alpha] anthracene (DMBA) and the formation of DMBA-DNA adducts. The DMBA-deoxyribonucleoside adducts from 7,8-BF-treated cultures had the same elution profiles on Sephadex LH-20 columns as those from cultures exposed to DMBA alone, but 7,8-BF-treated cultures contained smaller amounts of DMBA-DNA adducts per mg DNA. As the concentration of 7,8-BF was increased, the decrease in the amount of DMBA-DNA adducts per mg DNA was logarithmic with respect to the decrease in the amount of DMBA metabolized. The results suggest that more than one metabolic step is required for the binding of DMBA to DNA in hamster embryo cells.


The in vitro interaction of the carcinogens, 4-nitroquinoline-N-oxide and its metabolite 4-hydroxylaminoquinoline-N-oxide, with intact rat liver mitochondria.

The inhibition of rat liver mitochondrial respiration caused by rotenone, is relieved by the 2 carcinogens, 4-nitroquinoline-N-oxide (NQO) and its metabolite 4-hydroxylaminoquinoline-N-oxide (HAQO). Thus these agents cause reducing equivalents to circumvent the first coupling site of the respiratory chain. This is another example of the experimental confluence between oxidative phosphorylation and chemical carcinogenesis.


[Blood plasma differentiation of 11 domestic species by layer electrofocusing].

The study of the dispersion of plasma proteins of 11 species of Mammals by electrofocusing in thin layers, allows the characterisation of each species with the diagram of the migration of the proteins according to their isoelectric point. This fast and easy method may be preferred to immunological means and can be applied to the smallest sample forensic medicine.


[Evolution of median menigeal veins network in primates, from endocranial imprint patterns].

In the evolution of Primates, three main changes in the endocranial imprint pattern of the median meningeal veins are observed: 1. a gradual increase in the arrangement of the three-branched network: 2. a progression of the complexity of the anastomotic squaring; 3. the consecutive disappearnce of the retroarticular system (external jugular vein), which brings about an enlargement of the available space for the internal jugular system, and the ensuing increase in functional importance of the pterygoid plexus.


[Organization of contralateral and bilateral projections of corticospinal tracts in cats].

In Cats with a previous spinal lesion (C 6) saving only a lateral or ventral funiculus, a caudal horseradish peroxidase injection (C 8-Th 1) reveals that some cortical zones (lateral sigmoïd gyrus, medial part of the posterieur sigmoïd gyrus, medial part of the dorsal bank of the cruciate sulcus) send spinal projections, only contralaterally, by way of the dorsolateral funiculus, whereas other cortical zones (anterior sigmoïd gyrus, middle part of the posterior sigmoïd gyrus, lateral parts of the dorsal and ventral bank of the cruciate sulcus) send bilateral spinal projections by way of the ventral and dorsolateral funiculi.