Parametric studies of dorsal horn neurons responding to tactile stimulation. Dorsal horn neurons responding to tactile input were recorded in segments L3-S2 of unanesthetized, low-spinal cats. Single units were characterized with regard to receptive field (RF) location, RF size and shape, spontaneous discharge rate, central delay, and convergence of four tactile afferent types. 1. RF size increased from the toes to the calf region. 2. Length-width ratio increased from the toes to the calf and declined from the calf to the hip. 3. The relation between RF size and position on the limb was independent of segmental and laminar location of the neurons. 4. RF size was positively correlated with spontaneous discharge rate. 5. The relation between RF size and shape and RF position can be interpreted in terms of regional variations in the magnitude of the gradient of representation in the dorsal horn somatotopic map. 6. Central delay was negatively correlated with both RF size and rate of ongoing discharge. 7. There were no statistically significant differences among the laminae with respect to central delay, RF size or shape, ongoing discharge, or convergence combinations of the four tactile afferent systems. 8. Data presented were at variance with Wall's laminar cascading model for laminae IV-VI. Our results suggest that the model should be modified, at least to emphasize monosynaptic tactile input to all three of these laminae.


Progressive changes in kitten striate cortex during monocular vision. Following initial rearing in either total darkness or normal illumination, kittens at different ages were subjected to right-eye closure and various periods of vision through the left eye. After the period of monocular vision, single units in striate cortex were tested for visual responsiveness through each eye. A severe reduction in the proportion of units responsive to the deprived eye occurred over the first few days of monocular vision. Functional abnormalities were variably present after 1 day, marked after 2.5 and 3.5 days, and complete after 10 days. Monocular vision produced very much the same effect on ocular dominance of striate units, provided age and duration of suture were identical, regardless of whether kittens had received prior dark- or light-rearing.


Nonspiking interneurons in walking system of the cockroach. Intracellular recordings were made from the neurites of interneurons and motoneurons in the metathoracic ganglion of the cockroach, Periplaneta americana. Many neurons were penetrated which failed to produce action potentials on the application of large depolarizing currents. Nevertheless, some of them strongly excited and/or inhibited slow motoneurons innervating leg musculature, even with weak depolariziing musculature, even with weak depolarizing currents. Cobalt-sulfide-straining of these nonspiking neurons showed them to be interneurons with their neurites contained entirely within the metathoracic ganglion. Two further characteristics of these interneurons were rapid spontaneous fluctuations in membrane potential and a low resting membrane potential. One nonspiking neuron, interneuron I, when depolarized caused a strong excitation of the set of slow levator motoneurons which discharge in bursts during stepping movements of the metathoracic leg. During rhythmic leg movements the membrane potential of interneuron I oscillated with the depolarizing phases occurring at the same time as bursts of activity in the levator motorneurons. No spiking or any other nonspiking neuron was penetrated which could excite these levator motoneurons. From all these observations we conclude that oscillations in the membrane potential of interneuron I are entirely responsible for producing the levator bursts, and thus for producing stepping movements in a walking animal. During rhythmic leg movements, bursts of activity in levator and depressor motoneurons are initiated by slow graded depolarizations. The similarity of the synaptic activity in these two types of motoneurons suggests that burst activity in the depressor motoneurons is also produced by rhythmic activity in nonspiking interneurons. The fact that no spiking neuron was found to excite the depressor motoneurons supports this conclusion. Interneuron I is also an element of the rhythm-generating system, since short depolarizing pulses applied to it during rhythmic activity could reset the thythm. Long-duration current pulses applied to interneuron I in a quiescent animal did not produce rhythmic activity. This observation, together with the finding that during rhythmic activity the slow depolarizations in interneuron I are usually terminated by IPSPs, suggests that interneuron I alone does not generate the rhythm. No spiking interneurons have yet been enccountered which influence the activity in levator motoneurons. Thus, we conclude that the rhythm is generated in a network of nonspiking interneurons. The cellular mechanisms for generating the oscillations in this network are unknown. Continued.


Technetium-99m-pyrophosphate kinetics and imaging in metabolic bone disease. A study was undertaken to investigate the behavior of 99mTc-Sn-pyrophosphate complex in metabolic bone disease. Of clinical importance was the generalized increased periarticular bone accumulation of the radiopharmaceutical in osteomalacia and in combined osteomalacia and osteitis fibrosa as found in patients with chronic renal failure. The pattern in primary hyperparathyroidism was variable. There was no correlation between the initial rates of accumulation of the radiophosphate complex or its bone to soft-tissue uptake ratio at 5 hr when compared with the degree of osteomalacia and osteitis fibrosa. It is postulated that the 99mTc-Sn-pyrophosphate complex has greater affinity for immature collagen than the crystal surface.


Technetium-99m-pyrophosphate: studies in vivo and in vitro. In rats with induced rickets, the uptake of 99mTcO4 and 99mTc-pyrophosphate per gram of bone was increased as compared with weight-matched controls. However, the uptake of radioactive calcium and 32P-pyrophosphate was similar in both rachitic and control animals, suggesting that the 99mTc label conferred specificity and favored the rachitic lesions. Employing the rat tibia in an in vitro system, 99mTcO4 uptake was predominantly in the organic bone matrix; radioactive calcium, 32P-pyrophosphate, or 14C-diphosphonate uptake was mainly in the bone mineral; and 99mTc-pyrophosphate, 99mTc-diphosphonate, and 99mTc-polyphosphate were found in both mineral and organic phases. By removal of both mineral and polysaccharide and by using agents that altered the degree of collagen fibril cross-linking, evidence was obtained suggesting the 99mTcO4 and 99mTc-pyrophosphate are preferentially bound by immature collagen.


Distribution of 99mTc-Sn diphosphonate and free 99mTc-pertechnetate in selected soft and hard tissues. Because increased uptake of 99mTc-diphosphonate (ethane-hydroxy-1, 1-diphosphonate) occasionally occurs in the anterior neck region, the possible increased affinity of the diphosphonate bone-scanning agent for cartilage was investigated. In vivo scintigraphic studies and organ analyses from rats and rabbits injected with this bone scintigraphic agent were performed. Trachea-to-muscle uptake ratios were a high 45:1 in adult Sprague-Dawley rats and approached the femur-to-muscle ratio of 93:1. Technetium-99m-diphosphonate uptake was also increased, but to a lesser extent, in xiphoid cartilage, tendon, and ear cartilage; this was proportional to the calcium content of the organ. The thyroid showed a high affinity for free pertechnetate but not 99mTc-diphosphonate, providing further evidence that the increased neck uptake of this 99mTc-diphosphonate is due to tracheal, not thyroid activity. In addition, premedication of three patients with 200 mg of potassium-perchlorate did not block this neck uptake. Interpretation of scintigraphs performed with 99mTc-diphosphonate that show lesions in the cervical spine should take into account the potential for false-positive readings caused by this increased tracheal uptake.


Kinetics of 99mTc-labeled pyrophosphate and polyphosphate in man. Thekinetic of 99mTc-labeled pyrophosphate were compared with those of polyphosphate in ten patients in a combined study. Both agents cleared from the blood in a biexpoential fashion. The clearance half-time of Exponent I was the same for both and was shorter than the clearance half-time of Exponent ii. Urinary excretion of both agents was the same during the first hour but during the next 3 hr Tc-pyrophosphate cleared at a slightly more rapid rate, resulting in lower blood background radioactivity. Both agents were bound loosely to plasma proteins, mainly to globulin fractions. The sensitivity of lesion detection was similar for both. Excellent bone images were obtained with both agents. The images with Tc-pyrophosphatewere consistently superior owing to the low blood background and they took less time to accumulate an identical number of counts from identical regions. With the amount of 99mTc-complex used, no hyocalcemia or tetany was noted, nor was there any significant effect on 1-hr serum levels of inorganic phosphours and alkaline phosphatase. Four hours after injection, 9.5% of the dose of Tc-pyrophosphate was circulating in blood, 31.7% was excreted in urine, and the remaining 58.8% was taken up by bone and other tissues. The corresponding values with Tc-polyphosphate were 12.5% in blood, 29.0% in urine, and 58.5% in bone and other tissues. Among the soft tissues, the genitourinary system is most consistently visualized. It is concluded that both Tc-pyroposphate and Tc-polyphosphate are excellent skeletal-imaging agents and that Tc-pyrophosphate appears slightly superior to Tc-polyphosphate.


Radiocolloid scans in evaluating splenic response to external radiation. By use of 99mTc-sulfur colloid, splenic size as well as liver/spleen ratio of ratioactivity was determined after external radiation of the abdomen. In six patients receiving about 2,000 R whole-abdominal radiation, there was no atrophy of the spleen or abnormal distribution in the liver/spleen ratio of radioactivity (that is, the spleen was still functional). Serial studies in a 7-year-old boy with acute lymphoblastic leukemia in remission showed that 1,450 R splenic radiation did not result in any appreciable change in the length of the organ. In a woman with lymphosarcoma, a change in spleen size did not occur until a dose of 1,800 R was delivered. Another patient had apparently normal uptake of radiocolloid 5 years after 3,600 R. Hence the normal spleen and the spleen affected by other diseases may be far more resistant to external radiation than the spleen diseased with chronic myelocytic leukemia. Spleen scans can be useful in documenting the response of the organ to radiation.


Distributions of several agents useful in imaging myocardial infarcts. Myocardial cell death due to infarction is accompanied by an influx of calcium ion. The calcium ion seems to localize in crystalline structures that form within mitochondria and resemble hydroxyapatite. Based on this phenomenon 99mTc-stannous pyrophosphate has been successfully used to image myocardial infarcts within 24 hr of infarction and 1 hr following tracer administration both in dogs and patient volunteers. In this report, canine distribution studies of 99m-Tc-pyrophosphate are compared with similar studies with 99mTc-stannous polyphosphate, 99mTc-stannous 1, hydroxy-ethylidene-1, 1-disodium phosphonate diphosphonate), and 18F as sodium fluoride. Pyrophosphate polyphosphate, and diphosphonate are each potentially useful in myocardial infarct imaging but bone uptake of 18F occurs sufficiently early to prevent the use of this radionuclide in infarct scintigraphy.


Significance of renewal asymmetry in bone scans: experience in 795 cases. A retrospective study of 795 consecutive bone scans employing either 18F or 99mTc-pyrophosphate to evaluate the diagnostic value of renal asymmetry in such scans has been carried out. It is concluded that asymmetric renal images in bone scans convey relatively specific information regarding renal pathology, especially in the 99mTc-pyrophosphate studies.


Studies in type I glycogenosis: the paradoxical effect of ethanol on lactate. Paradoxically, ethanol, which raises lactate in normal individuals, lowers the elevated levels of lactate in patients with Type I glycogenosis. We found that, although lactate levels fell, pyruvate proportionately declined even more, resulting in an increased L/P ratio which indicates that, as in the normal, the oxidation of ethanol had generated NADH. In type I glycogenosis, the increased level of pyruvate-lactate derives from glycogenolysis. We found that, despite continued glycogenolysis, ethanol had caused less pyruvate-lactate to form. The effect of an increased NADH/NAD+ ratio on the flow of carbon through the Embden-Meyerhof pathway could account for the finding, presumably by its effect on the oxidation-reduction couples with diversion of carbon toward formation of triglyceride rather than pyruvate-lactate.


Glycerol intolerance in a child with intermittent hypoglycemia. A 3-yr-old boy was investigated for numerous episodes of fatigue, irritability, pallor, and sweating, which began at 11 mo of age, when he had an episode of symptomatic hypoglycemia with ketonuria. He had euphoria, mental confusion, drowsiness, nausea, and vomiting 1-5 hr after oral administration of glycerol in doses of 0.5-1.0gm/kg. Orally administered MCT (1 gm/kg) had similar effects. On one occasion, oral glycerol also provoked hypoglycemia, as had a 16 1/2 hr fast. Intravenously administered glycerol (0.09 gm/kg) induced an immediate loss of consciousness from which he recovered spontaneously after 30 min; there were no changes in blood glucose values. Intravenously administered fructose (0.25 gm/kg) was tolerated normally. Leukocytes showed normal activities for FDPase, glycerol kinase, and glycerol phosphate dehydrogenase. The restriction of dietary intake of fat has been associated with a marked improvement in physical and mental activities. These observations suggest a unique, yet undifined intolerance to glycerol, which suggest caution in the diagnostic use of glycerol in the investigation of hypoglycemia as well as in the therapy of increased intracranial or intraocular pressure.


Multiple cerebral abscesses complicating hepatopulmonary amebiasis. A 3-year-old boy with hepatopulmonary amebiasis treated with chloroquine phosphate for 20 days subsequently was found to have three brain abscesses and responded to prolonged combined therapy with metronidazole and chloroquine phosphate. Cerebral amebiasis should be considered in any patient with amebic dysentery, hepatic abscess, or pulmonary involvement in whom signs or symptoms of central nervous system involvement appear. A prolonged course of appropriate therapy with periodic brain scanning for evidence of improvement is suggested.


Endocrinological findings in sterile pink-eyed mice. Mice homozygous for  rho-25h, a mutation at the pink-eye locus, are small, nervous and male sterile, with abnormal sperm morphology: females have very low fecundity. Evidence is presented that in both sexes the pars nervosa of the rho-25h/rho-25h pituitary contains degenerating nerve axons, that the hypothalamus has a reduced binding capacity for oestradiol-17 beta and that thyroid iodine binding is reduced. No changes were observed in the anterior pituitary. In rho-25h/rho-25h females, the uterus is threas-like and unresponsive to oestradiol, and the ovaries are small with many abnormal follicles. Possible modes of action of the rho locus are duscussed with reference to this and pre-existing data on the rho phenotype.


A review of tumor antigens in gynecologic malignancies. Evidence for the presence of tumor-associated gynecologic tumor antigens from our laboratory and others is presented. Tumor-associated antigens have been described for invasive carcinoma of the cervix and mucinous carcinoma of the ovary with cross-reacting antigens in malignant vulvar tissue. The provocative finding of a cross-reacting antigen in malignant cervical, vulvar and ovarian tissue raises the possibility of a common etiologic agent. The hope of the clinician is to utilize the tumor antigens in diagnosis and therapy. However, it is necessary to caution against overzealous acceptance so the carcinoembryonic antigen story is not repeated.


Minimal growth requirements for Clostridium perfringens and isolation of auxotrophic mutants. The minimal growth requirements for two strains of Clostridium perfringens were defined, and both synthetic and semisynthetic plating media were developed. Plate counts of the wild-type strains on both of these minimal media were equivalent to those on complex media. A number of auxotrophic mutants of each strain were isolated, and their phenotypes were defined.


Recovery of murine leukemia virus from large volumes of freshly harvested culture fluids by using a single density gradient. Concentration and purification of murine leukemia virus for use as complement fixation antigens was accomplished by using a single density gradient in the model RK ultracentrifuge.


Septic and crystalline joint disease. A simultaneous occurrence. Bacteria and crystals were simultaneously recovered from the synovial fluid of two patients with acutely inflamed joints. The bacteria were initially identified on Gram stain and subsequently grown on supplemented peptone broth, and the crystals were readily identified by polarizing microscopy. Although cause-and-effect relationship between these two types of arthritis cannot be established, the need for thorough examination of synovial fluid is evident.


Differentiation of two abnormalities in thyroid peroxidase causing organification defect and goitrous hypothyroidism. Clinical and laboratory evaluations are reported on two patients with congenital goiter and hypothyroidism due to iodide organification defect. In one patient, a 31-year-old white male with severe mental retardation, administration of perchlorate caused discharge of 69% of the radioiodine accumulated in the thyroid gland. Thyroid tissue had negligible peroxidase activity in the tyrosine-iodinase, triliodide, and guaiacol assays. Preincubation of subcellular fractions with hematin restored activity. The restored enzyme was labile to high concentrations of H2O2 (5.6times 10-4 h2o2 produced inhibition in the triiodide assay). Heating of the enzyme for 5 min at 46 degrees C produced 50% inactivation, while higher temperatures were required to half-inactivate normal peroxidases. This case represents a second example of the "peroxidase apoenzyme-prosthetic group defect" causing congenital goiter. The second patient, an example of the "deficient peroxidase defect," was a 10-yr-old girl with 35% discharge of thyroidal radioiodine by perchlorate. Peroxidase activity in the goiter tissue was quantitatively decreased (10%-20% of normal values) but kinetically normal with respect to apparent Km for H2O2. Hematin had little effect on the enzyme. Peroxidase activity had abnormal subcellular distribution, since pellets sedimenting between 39,000 and 105,000 g contained most of the activity. Normal thyroglobulin was observed in the thyroid gland of the patient. Two distinct defects of the peroxidase system can produce congenital goiter by limiting organification of iodide.


Primary Epstein-Barr-virus infections in acute neurologic diseases. Infectious mononucleosis has been associated with Guillain--Barré syndrome, Bell's palsy, meningoencephalitis and transverse myelitis. Since it is not known that many children with infectious mononucleosis do not develop heterophil antibodies, we looked for evidence of current or recent Epstein-Barr virus infection in young patients with these neurologic diseases by using serodiagnostic procedures for detection and titration of antibodies to various antigens related to Epstein-Barr virus. Seven of 24 cases with Guillain-Barre syndrome and three of 16 with facial palsy were definitely associated with primary infection with Epstein-Barr virus as were two cases each of the other two neurologic diseases. Only one of these patients had obvious clinical infectious mononucleosis, and only a few demonstrated heterophil agglutinins. It is evident that the virus must be considered in the diagnosis of various acute neurologic diseases affecting children and young adults, even in the absence of heterophil-antibody response or other signs of infectious mononucleosis.


Long-term steroid therapy in patients with adrenogenital syndrome. Fifteen years' experience with the adrenogenital syndrome at Duke University Medical Center is reviewed. Twenty-six patients were diagnosed before age 2. Of these, 20 had the salt-losing syndrome. The complications encountered and the results obtained in terms of growth, developemnt, and fertility are discussed. Delay in diagnosis and inappropriate gender assignment continue to be a problem.


Adrenal virilism due to 21-hydroxylase deficiency in the postmenarchial female. Five new patients with postmenarchial onset of adrenal virilism related to 21-hydroxylase deficiency are presented. Diagnostic criteria are reviewed. Four pregnancies resulting in 3 term infants occurred after instigation of therapy. Eleven patients similar in clinical presentation are reviewed for comparison. Findings of value in distinguishing these two groups of patients are discussed.


Paget's disease of the vulva: a clinicopathologic study of eight cases. Clinicopatholigic findings in 8 patients with Paget's disease of the vulva are presented with a discussion if its pathogenesis. In 7 cases the disease was intraepithelial, and in 1 it was accompanied by underlying adenocarcinoma. The treatment of choice is surgical. The importance of early diagnosis, adequate excision, and close followup is emphasized. In 2 cases, the disease was associated with carcinoma of the breast and in 1 case with carcinoma of the liver.


Dynamic scintigraphy of primary and secondary malignant intracranial neoplasms. Dynamic images and arteriograms were reviewed in patients with proved primary and secondary malignant intracranial neoplasms. In 44% of the patients with glioblastomas and 23% of those with metastatic lesions, radioactivity appeared early in the dynamic imaging sequence. This was related to the presence of arteriovenous shunting on arteriography rather than to a tumor stain. Neither characteristic dynamic imaging patterns nor rapid arteriovenous shunting was seen in patients with astrocytomas.


Experience with 111-In-chloride scanning in patients with focal defects on 99-mTc-sulfur colloid liver scans. Thirty-nine patients with focal defects on the technetium liver scan were rescanned using 111-In chloride. Of 20 patients with hepatic malignancy, 11 had positive indium scans. None of the 19 with focal cirrhotic fibrosis had a positive indium scan although 5 of these had primary tumor. Thus, a positive indium scan suggests that the defect is malignant. A negative indium scan is less helpful, failling to distinguish between neoplasm and focal cirrhosis. Positive uptake in an extrahepatic primary neoplasm and negative uptake in the liver suggest that the hepatic lesion is not neoplastic.


Irradiated lung cancer. An autopsy analysis of spread pattern. The spread pattern was studied in 48 autopsied cases of irradiated, apparently localized lung cancer. Disease was found in the upper abdomen in 52%, in the brain in 8.7%; in the bones in 20.8%; and confined to the thoracic cavity in 27%. Radical irradiation controlled 38% locally. The pattern of metastasis in these irradiated lung tumors did not seem to differ from that of unirradiated lung cancers reported elsewhere.


Carcinoma associated with irradiation of the immature breast. In two women who had undergone radiation therapy at ages one and eleven years, respectively, carcinoma in a hypoplastic breast developed. Each patient was treated by radical mastectomy. One patient is presently being followed without evidence of metastases; the other died of sarcoidosis following thoracotomy. A history of irradiation, hypoplasia and microscopic findings compatible with radiation changes suggests that these two cases of carcinoma may be radiation-induced.


The inheritance of mild Type II hyperlipoproteinaemia. The distribution of plasma total and low density lipoprotein cholesterol concentration was investigated in 158 first degree relatives (105 progeny, 53 siblings) of 55 index patients with mild type II hyperlipoproteinaemia. No evidence of bimodality was found. Our findings support the view that mild type II hyperlipoproteinaemia is inherited in a polygenic manner.


Adenosine 3',5'-monophosphate analogs promote a circular morphology of cultured Schwannoma cells. Dibutyryl adenosine 3',5'-monophosphate (dibutyrl cycle AMP) and 8-bromo cyclic AMP stimulated cells cultured from a rat Schwannoma to change their morphology from irregularly shaped to flattened circular and hollow circular forms within 30 minutes. The change in shape was specifically effected by analogs of cyclic AMP and cyclic AMP elevating agents, was reversible after removal of these additives, and was prevented by vinblastine and cytochalasin b, but was not affected by actinomycin D or cycloheximide.


Reduced incidence of spontaneous mammary tumors in C3H/He mice after treatment with polyadenylate-polyuridylate. The effect of treatment with the double-stranded polynucleotide complex polydenylate with polyuridylate [poly(A) with poly(U)] on tumor development in C3H/He mice was evaluated. Poly(A) with poly(U) was injected in newborn females, and mice were observed for 380 days. During this experimental period 42 percent of treated mice developed tumors, while the incidence in the control group was 63 percent. This difference was statistically significant.


Collagenase production by lymphokine-activated macrophages. Macrophages incubated with products (lymphokines) secreted by stimulated spleen cells produced collagenase. Active lymphokines were obtained both from mitogen- and antigen-stimulated lymphocytes. These observations suggest that the degradation of collagen in chronic inflammatory lesions may be caused by macrophage collagenase.


Endogenous cyclic adenosine monophosphate in tissues of rabbits fed an atherogenic diet. Rabbits fed a high cholesterol diet experienced a significant increase in plasma adenosine 3,5-monophosphate (cyclic AMP), which was simultaneous with the increase in plasma cholesterol. The content of cyclic AMP in atherosclerotic lesion areas of rabbit aortic intima-media was significantly higher (0.24 picomole per microgram of DNA) than that in adjacent nonlesion areas or in aortic intima-dedia from control animals (0.09 picomole per microgram of DNA). The cyclic AMP content of heart, liver, skeletal muscle, and diaphragm showed no significant elevation in animals fed cholesterol.


Characterization of a nuclear receptor for testosterone in rat bone marrow. In vitro testosterone binding was studied in a nuclear extract (0.15 M KC1) from rat bone marrow using the charcoal assay. The nuclear extract contains binding sites saturable at low concentration of testosterone (1 times 10-8M). From a Scatchard plot of the data obtained it is concluded that a single class of receptor sites is involved in the binding of testosterone. Studies indicated that the testosterone-nuclear complex has a dissociation constant (Kd) of 5.9 times 10-9 M binding 9 times 10-14 moles/mg nuclear protein. 5alpha- and 5beta-dihydrotestosterone compete poorly with testosterone for binding sites in the nuclear component while estradiol -17beta does not compete at all. The protein nature of the receptor was demonstrated and an isoelectric point (pI) of 4.9 was determined for the complex by the use of isoelectric focusing.


A manganese catalyzed covalent binding of estradiol - 17 beta to cytosol proteins of rabbit uterus. Studies of the temperature sensitivity of estradiol receptor binding in rabbit uterine cytosol have revealed the existence of an enzyme which catalyzes the covalent binding of estradiol to cytosol proteins. A fraction, prepared by chromatography on Biogel P-200 and incubated at 37 degrees C in the presence of Mn++, exhibited a time-dependent, temperature-sensitive, oxidative binding of estradiol not seen in the crude cytosol preparation. Although the activity of this enzyme was shown to be independent of estradiol binding by the high affinity estrogen receptor, its presence may complicate studies of estrogen receptor action which involve the use of elevated temperatures.


Chemical action of influenza virus on the red blood cell membrane. II. Action on membrane lipids and amino acids. Haemagglutination by influenza virus strains PR8 and Lee as well as treatment of red blood cells by periodate produced numerous and manifold changes in the lipids and the amino acid composition of the erythrocytes. The changes induced by the influenza strains differed from each other and from those induced by periodate. Alterations in the membrane lipids observed after haemagglutination by heat treated strain PR8 and by its native form were identical.


The effect of supraoptimal temperature on the multiplication of different cytomegalovirus strains. The virion synthesis by five human cytomegalovirus (CMV) strains in human embryonic fibroblast cultures was stopped by incubation of the infected cultures at 40 degrees C. At this temperature the antigens appeared diffusely filling the nucleus and the cytoplasm. The blocking effect of the elevated temperature was exerted in the same period of the reproduction cycle as the inhibitory effect of cytosine arabinoside (ara-C). In cell cultures infected with CMV and incubated first at 40 degrees C, then at 37 degrees C, the synthesis of infectious virus started again, thus the abortive cycle developed at 40 degrees C was reversible. The inhibition of virus multiplication cannot be attributed to the thermosensitive events in the normal function of the host cell.


Chemical action of influenza virus on the red blood cell membrane. I. Action on membrane neuraminic acids. The chemical action of influenza virus strains PR8 and Lee and of periodate on the neuraminic acid containing lipids of human red blood cells has been investigated by thin layer chromatography. The results showed that haemagglutination by the two strains as well as periodate treatment induced manifold and profound chemical alterations in the neuraminic acid-containing lipids. The changes produced by strains PR8 and Lee differed from each other and from those caused by periodate. Alterations in the neuraminic acid-containing lipids induced by heated strain PR8 were identical with those obtained after haemagglutination by the native virus, indicating that virus neuraminidase is not inactivated by heat. Thus, differences in the haemagglutinating ability between native and heated virus are not due to enzyme inactivation.


X-ray diffraction from micrograms of quartz on filters using a rotating anode. X-ray diffraction data from samples of 20, 60 and 100 mug quartz on poly-vinyl chloride membrane filters have been collected using a rotating anode x-ray source. These data suggest that 17 mug of quartz or possibly less could be measured directly on 35 mm diameter filters without any type of pretreatment.


Identification of nonfermentative gram-negative bacteria in the clinical laboratory. A simplified, concise scheme was developed for the identification of nonfermentative, gram-negative bacteria which have most frequently been reported in the literature as definite or possible agents of human disease. These organisms included apyocyanogenic Pseudomonas aeruginosa, P. fluorescens, P. putida, P. stutzeri, P. maltophilia, P. putrefaciens, P. cepacia, P. alcaligenes, FLAVOBACTERIUM SPECIES, Bordetella bronchiseptica, Acinetobacter anitratum (Herellea vaginicola), A. Iwoffi (Mima polymorpha), Moraxella species, Alcaligenes odorans and Alcaligenes species. The tests used for identification included production of cytochrome oxidase, amylase, deoxyribonuclease, gelatinase, urease and Beta-galactosidase; motility; oxidation of one per cent glucose and ten per cent lactose; fluorescence; indole, hydrogen sulfide and nitrogen gas production; denitrification of nitrites; growth at 42C; penicillin sensitivity and production of an aromatic odor and greenish discoloration on blood agar. Using this scheme, 85 per cent of 243 isolates (unknowns and reference strains) were identified to genus and species. Of the 15 per cent remaining, 11 per cent were identified as alkaline organisms and four per cent were unidentifiable.


Hyperkinetic children: the use of stimulant drugs evaluated. Use of stimulant drugs in the treatment of hyperkinetic children is reviewed briefly. While drugs appear to be effective in "normalizing" behavior, too little attention has been paid to potential side effects. There is need for a broader program of clinical evaluation, assessing drug effects not only on behavior, but on appetite and the cardiovascular system, and taking into account the sociological impact of drug use by children.


Electron microscope studies of experimental Entamoeba histolytica infection in the guinea pig. I. Penetration of the intestinal epithelium by trophozoites. Germ-free guinea pigs were inoculated intracecally with Entamoeba histolytica and the enteric flora derived from a human patient with acute amebic colitis. Aanimals were killed at post-inoculation intervals of 7 to 12 days. The mode of penetration of cecal epithelium by the ameba was examined by light and electron microscopy. The following sequence was reconstructed from numerous individual observations. When the amebae were in moderately close proximity to the brush, border, the microvilli became shortened, irregular, and sometimes disappeared. Dense material was observed between the amebae and microvilli. When the ameba was very close to the epithelium the apical portion of epithelial cytoplasm projected into the lumen contacting the organism, thus becoming detached from adjoining cells. This produced spaces between epithelial cells through which amebae invaded interepithelial spaces. Initially the ameba penetrated the interglandular epithelium. Later, it penetrated equally the glandular and interglandular epithelium barrier. There were marked alterations of cytoplasmic components of epithelial cells. Polymorphonuclear leukocytes migrated into the epithelium filling these spaces; these often showed a variety of degenerative processes. Amebae, utilizing their pseudopodia, moved further through the intercellular spaces and reached the lamina propria.


Chemotherapy of experimental Echinococcus granulosus infection. Trials in CF1 mice and jirds (Meriones unguiculatus). Mice and jirds with experimental secondary hydatidosis (Echinococus granulous) were treated at various post-infection periods with eight different drugs; iodinized oil of thymol, ethyl-N-dimethyl ether of thymol fumarate, chloroguanide, rifampin, pentamidine isethionate, amphotericin B, suramin, and methotrexate. In initial experiments with methotrexate, mice treated after 99 days of infection had significantly fewer infections at necropsy than did controls (33% and 100%, respecitively). However, in subsequent studies at three dose levels of methotrexate in mice 10 days after infection and in mice and jirds 240 days post infection, no significant differences were found in the percentage of animals infected, cyst size, or number between treated and control animals. The other drugs tested were similarly without effect under the experimental conditions used.


Swine buffy coat culture: an aid to the laboratory diagnosis of hog cholera. A 2-step technique for the isolation of hog cholera (HC) virus consisting of an initial culture on buffy coat (BC) cultures and subinoculation to a pig kidney cell line (PK-15) was described. By this technique, HC virus was confirmed in specimens from 65 herds in which the conventional cell culture isolation technique failed. The herds were located in 20 states and Puerto Rico. Specimens from 29 of the 65 herds were inoculated into specific-pathogen-free (SPF) pigs. Hog cholera virus was recovered from 27 of the test pigs. The 2 pigs from which virus was not recovered had signs of acute infection and, on necropsy, had gross lesions of HC infection.


The Tecumseh study of respiratory illness. VIII. Acute infection in chronic respiratory disease and comparison groups. Individuals with chronic lung disease and their families were selected from the Tecumsch community along with similarly selected families as comparison groups and studied for 1-year periods. Occurence of acute respiratory illness was ascertained weekly by telephone and calculated as an annual rate. Persons with chronic bronchitis not only experienced more acute lower respiratory illness than healthy comparison subjects, but total illness rates were somewhat higher as well. Infection rates were determined from blood samples taken 3 times from each participant during the surveillance year. Antibody tests were performed for respiratory syncytial virus, para-influenza virus types 1, 2, and 3, influenza types A and B, coronavirus OC43, Mycoplasma pneumoniae, and Haemophilus influenzae. Differences in serologic infection rates among the subgroups of the population were similar to those seen in the clinical data, with more frequent infection among those with bronchitis than among the comparison subjects. This finding indicates that some degree of increased susceptibility to actual infection existed among those individuals with bronchitis. Influence of smoking on illness and infection rates was also examined. Infections were, in general, more frequent in smokers than in nonsmokers, but illness rates were reversed, suggesting that perception of disease differed in the 2 groups. Rates of illness and infection of other adults in the families of the index individuals with bronchitis were not influenced by the higher rates seen in the index individuals; however, it was of interest that children of persons with bronchitis did have somewhat higher rates of infection than children of comparison subjects.


Roentgenographically atypical Pneumocystis carinii pneumonia. Two patients with renal transplants were admitted for evaluation of fever. During the course of hospitalization both had hectic fever and arthralgia. Pulmonary symptoms were absent or minimal. In one patient the admitting chest roentgenogram was entirely normal and in the other only a small focal abnormality was apparent. Hypoxemia occurred even in the presence of normal or nearly normal chest roentgenograms. The diagnosis of Pneumocystis carinii infection was made by bronchoscopic brush biopsy in both patients. Therapy with pentamidine isethionate was successful. It is suggested that in patients with renal transplants and in others with similar immonosuppression, even with a normal chest roentgenogram, Pneumocystis carinii infection be considered as the cause of a fever of unknown origin. This should be evaluated initially with blood gas studies; if these are abnormal, further studies, including biopsy, are justified.


The effects of proteolytic enzymes on the tensile strength of human lung. Human lung tissues were exposed to proteolytic enzymes to determine the effects on tensile strength and to clarify the relationship between tensile strength and the amounts of collagen and elastin in the tissue. Elastase and papain depleted the tissue of elastin but failed to alter tensile strength. Trypsin had no effect on tensile strength, or on collagen and elastin content. collagenase lowered tensile strength and reduced the amount of collagen in the tissue. The findings with collagenase were in agreement with measurements in control tissues that showed a direct relationship between tensile strength and collagen content. These results confirm collagen as the principal determinant of the tensile strength of human lung.


Synthesis of factor VIII antigen by cultured human endothelial cells. Endothelial cells have been isolated from freshly obtained human umbilical cords by collagenase digestion of the interior of the umbilical vein and grown in tissue culture. These cells have been identified by morphologic and immunologic criteria. It has been demonstrated that these cultured human endothelial cells synthesize and release factor VIII antigen but not factor VIII clot-promoting activity.


Ultrastructural studies of regenerating spines of the sea urchin Strongylocentrotus purpuratus. II. Cell types with spherules. The fine structure of spherulecytes, cell types with large, intracellular membrane-bound vacuoles termed spherules, was investigated in regenerating tips of spines of the sea urchin Strongylocentrotus purpuratus. Two categories of cell types were observed: red spherulecytes and colorless spherulecytes. Red spherulecytes were represented by a single cell type, the eleocyte, while colorless spherulecytes consisted of three morphologically distinct cell types termed morula cells, granulocytes, and vacuolecytes. Eleocytes and morula cells were distributed in both the epidermis and dermis, while granulocytes and vacuolecytes were present only in the dermis. After processing for light and electron microscopy, the spherules of eleocytes typically appeared empty, having lost their content of the red pigment, echinochrome. In contrast, the spherules of morula cell, granulocytes, and vacuolecytes enclosed a variety of granular and other material. The cell types reported in this paper resembled, to various degrees, spherulecytes in the coelomic fluid of echinoids described by other investigators.


Cellular structure and function in normal and neoplastic lymphoid cells. In general, the ultrastructure of normal lymphocytes and plasma cells reflects their functional state. Similarly, in pathologic conditions, ultrastructural abnormalities may reflect specific functional derangements of the cells. The identification of some structural abnormalities may be clinically useful, even though their origin and biochemical composition is still obscure.


Radionuclide angiography in juvenile angiofibroma of the nasopharynx. Radionuclide angiography was performed by intravenous injection of 99mTc-pertechnetate and recorded by serial gamma camera scintiphotographs. Its usefulness as a prebiopsy or preoperative procedure is demonstrated by two case reports of angiofibroma of the nasopharynx and it was also found to be useful for radiation therapy follow-up assessment. Radionuclide angiography offers a convenient and safe method to assess the vascularity of a mass and by this technique the surgeon may be alerted to the possibility of severe postbiopsy hemorrhage and also the occasional dramatic complications of radiological carotid angiography can be avoided.


Sleep disturbance in schizophrenia. A revisit. Rapid eye movement (REM) sleep mechanisms may play a role in the pathophysiology of schizophrenia, but results have been inconclusive and studies of sleep in schizophrena have been deficient in identifying, comparing, and differentiating between subcategories of schizophrenia. Twenty-nine hospitalized, drug-free schizophrenics were divided into three subgroupsācute, latent, and schizoaffective. The REM intensity measures and REM latency were found to differentiate significantly the schizoaffective group. Sleep-continuity indexes separated the latent and acute groups. Seven patients who later required treatment with tricyclic an tidepressants had base line REM latencies significantly lower and hospitalizations significantly more prolonged than the patients who did not require antidepressants. Sleep measurements may thus identify diagnostic subgroups of schizophrenia as well as predict which schizophrenic depressive syndrome or a concurrent affective syndrome.


Glomus tumors in the head and neck: III. Analysis of clinical manifestations. Seventy-five patients with glomus tumors in the head and neck had a 37% incidence of cranial nerve paralysis and a 14.6% incidence of intracranial extension. Jugular foramen syndrome is associated with 50% and hypoglossal nerve involvement with 75% posterior fossa tumor invasion. Horner's syndrome is associated with 50% middle cranial fossa tumor invasion. The incidence of central nervous system (CNS) involvement with cranial NERVE PARALYSIS (NOT INCLUDING VII nerve) is 52%. Otologic findings and VII nerve paralysis did not correlate with tumor resectability, CNS extension, and prognosis.


Congenital herpetic keratitis.

Congenital herpetic keratitis. An infant, the first-born of twins, delivered by cesarean seciton without prior rupture of the membranes, was noted to have an advanced epithellal and stromal keratitis at berth. Herpes simplex, type 1, was cultured form the cornea and from skin vesicles which developed subsequently. The advanced nature of the lesion indicates that it was acquired in utero. This appears to be the first reported case of congential herpetic keratitis.


Amyotrophic lateral sclerosis. Inclusion bodies in a case of the classic sporadic form. Postmortem light and electron microscopic studies of a 52 year old black male who died 17 months after the onset of upper and lower motor neuron signs showed: (1) degeneration of cortico-spinal tracts, (2) loss of spinal neurons and gliosis and (3) cellular inclusions with neurotubules, neurofilaments and granular material. Although these cellular inclusions resemble Lafora bodies, they differ in that, to the authors' knowledge, the latter were not reported to have microtubules. Review of the literature revealed no previous report of these inclusions in cases of amyotrophic lateral sclerosis. Tissue cultures of cord, hindbrain and cerebrum did not show cytopathic effect during a three-week observation period.


Centronuclear myopathy: histochemistry and electron microscopy. Report of two cases. In two new cases of centronuclear myopathy, histochemical findings included failure to differentiate fiber type with oxidative enzymes, the presence of core glycogenosis, and core acid phosphatase activity. Fluorescence microscopy demonstrated autofluorescent  lipochrome in fiber centers and nonspecific fiber injury in some immunofluorescent preparations. Electron microscopical findings included the observation of unusually small myofibrils arrayed between central nuclei.


Functional differences in protein synthesis between rat liver tRNA and tRNA from Novikoff hepatoma. Synthesis of ovalbumin in fragmented oviduct magnum explants of immature, estrogen-stimulated chicks has been studied in the presence of exogenous tRNA. tRAN from Novikoff hepatoma specifically inhibited ovalbumin synthesis, determined by precipitation with antisera. In addition, the major protein(s) synthesized in the presence of hepatoma tRNA had higher electrophoretic mobility than ovalbumin, as shown by sodium dodecyl sulfate polyacrylamide gel electrophoresis. tRNAs from rat liver, rooster liver, and hen oviduct did not affect ovalbumin synthesis, although oviduct tRNA is stimulatory during the earlier stages of estrogen stimulation.


Oligonucleotide sequences of pancreatic and T1 ribonuclease digests of 5S ribosomal RNA from mouse cells. Oligonucleotides produced by complete pancreatic and T1 RNase digestion of 5S ribosomal RNA from a mouse hepatoma, MH 134, have been separated with two-dimensional electrophoresis and their nucleotide sequences determined. Except for the presence of a 5'-terminal diphosphate, these nucleotide sequences were identical with those of KB cells, confirming the identity of the primary structure of 5S RNA between these animals. Both oligonucleotide patterns produced with these enzymes from 5S RNA of the liver were also identical with those of the hepatoma. All these agree with the strong conservation of 5S RNA genes in animal species. However, when 5S ribosomal RNA was extracted from ribosomes which were prepared from microsomal pellet, pancreatic RNase digest contained two trinucleotides (A-G-Cp and G-A-Cp) that were not found in 5S RNA prepared with a one-step procedure. It was concluded that different isolation procedure might indeed cause artifactual fragments on enzymatic digestion due to internal nicks produced during isolation. The significance of 5'-terminal diphosphate in relation to the biosynthesis of 5S ribosomal RNA is also discussed.


Inhibition by cyclic AMP and dibutyryl cyclic AMP of transport of organic acids in kidney cortex. 1. Cyclic adenosine 3',5'-monophosphate and N-6-2'-O-dibutyryl cyclic adenosine 3',5'-monophosphate decrease the initial entry rate and the steady-state uptake of p-aminohippurate and uric acid by rabbit kidney cortex slices. 2. N-6-2'-O-Dibutyryl adenosine 3'-5'-monophosphate inhibits the tubular transport of p-aminohippurate competitively. 3. Isoproterenol, known to increase cyclic nucleotide concentration of the cortical tubules by activation of adenyl cyclase, decreases p-aminohippurate transport. Antidiuretic hormone which is known to stimulate only medullary adenyl cyclase has no effect on p-amino-hippurate uptake by cortical slices. 4. Theophylline, which inhibits cyclic nucleotide phosphodiesterase and, therefore, enhances the cellular accumulation of endogenous cyclic nucleotide, depresses p-aminohippurate transport.


Effect of polymyxin B on liposomal membranes derived from Escherichia coli lipids. The specificity of the action of polymyxin B was studied using liposomes as a model membrane system. Liposomes prepared from total lipids of Gram-negative bacteria Escherichia coli, a mixture of purified E. coli phosphatidylethanolamine and cardiolipin and a mixture of phosphatidylethanolamine and phosphatidylglycerol, were extemely sensitive to polymyxin while those prepared from lipids of Gram-positive bacteria Streptococcus sanguis, lipids of sheep erythrocyte membranes, mixtures of egg lecithin and negatively charged amphiphatic molecules, were less sensitive to the action of the antibiotic. Cholesterol was shown to suppress the polymyxin-induced response in liposomes.


Magnetic resonance studies on the mitochondrial divalent cation carrier. Measurements of water proton spin relaxation enhancements (epsilon) can be used to discriminate high-affinity binding of Mn-2+ or Gd-3+ to biological membranes, from low-affinity binding. In rat liver mitochondria, epsilon b values of approx. 11 are observed upon binding of Mn-2+ to the inner membrane, while internal or low-affinity binding remains invisible to this technique. Energy-driven Mn-2+ uptake by liver mitochondria results in the subsequent decay of epsilon. Comparison of epsilon with the initial velocity of Mn-2+ uptake in rat liver mitochondria reveals a linear correlation, which holds at all temperatures between 0 degrees C and 40 degrees C, regardless of the mitochondrial protein concentration. Consequently, enhancement appears to reflect the binding of Mn-2+ to the divalent cation pump. Binding of Mn-2+ to blowfly flight muscle also results in substantial epsilon, which is associated with the glycerol-1-phosphate dehydrogenase instead of divalent cation transport. Consequently, no decay in epsilon due to uptake occurs after Mn-2+ is bound. Lanthanide ions are also bound and transported by mitochondria. Addition of Gd-3+ to pigeon heart or rat liver mitochondria results in epsilon b approximately equal to 5-6, which decays with similar kinetics in both systems. The uptake velocity of Gd-3+ in rat liver mitochondria is about 1/6 the rate with which Mn-2+ is transported. Lanthanides also diminish epsilon due to the addition of Mn-2+, and greatly retard the Mn-2+ uptake kinetics. The presence of carbonylcyanide-p-trifluoromethoxyphenylhydrazone depresses epsilon upon addition of Mn-2+ or Gd-3+ and also uncouples energy-driven uptake. On the other hand, prolonged anaerobic incubation in the presence of antimycin and rotenone exhausts the mitochondria of their energy stores, blocks the uptake of Mn-2+, but does not affect epsilon significantly. Evidently, the uncoupler-induced disappearance of divalent cation binding sites is not the result of "de-energization". Measurements of epsilon at several NMR frequencies indicate a correlation time (tau b) for carrier-bound Mn-2+ in rat liver mitochondria between 20 ns and 4 ns as one varies the temperature between 10 degrees C and 30 degrees C. The 13 Kcal/mole activation energy for tau b suggests that the 11 ns time constant at room temperature represents the movement of the Mn-11-carrier comples. On the other hand, tau b is probably approx. 100 times too short to represent the rotational motion of a carrier protein. Apparently, Mn-2+ binds to a small arm of the carrier which moves independent


Androgen receptors: relationship to growth response and to intracellular androgen transport in nine variant lines of the Shionogi mouse mammary carcinoma. Aspects of the biological significance of androgen receptors have been studied in nine variant lines of the Shionogi carcinoma, two of which are androgen dependent and seven of which are autonomous. The dependent lines, and two of the seven autonomous lines, contain androgen receptors; this finding demonstrates that the presence of receptors is not an accurate marker of hormonal dependence in vivo. Since the ability to transport androgens into the nucleus, as judged from the relative maximal rates of transport, is virtually restricted to dependent and autonomous lines which possess cytoplasmic receptors, it is clear that such receptors may play a role in regulating the intranuclear concentration of androgens. The absence of cytoplasmic receptors and the comparative lack of perceptible transfer of androgens across the nuclear membrane are features peculiar to the autonomous condition.


Altered hormone control of cyclic AMP formation in isolated parenchymal liver cells from rats treated with 2-acetylaminofluorene. The hormone control of cyclic AMP-formation in isolated parenchymal liver cells from rats fed the carcinogen 2-acetylaminofluorene (0.025% for 4-8 weeks) was studied. The cells from the carcinogen-treated animals responded much more strongly to adrenergic agents than cells from control animals, while no significant difference was found for the glucagon effect. Of the adrenergic substances studied, the order of potency was isoprenalin larger than or equal to adrenalin larger than phhenylephrine; stimulation was blocked by propranolol, but not by phentolamine. The effects of supramaximal concentrations of isoprenalin and glucagon were not additive.


Stimulation of cyclic adenosine 3':5'-monophosphate and corticosterone formation in isolated rat adrenal cells by cholera enterotoxin. Comparison with the effects of ACTH. 1. The production of cyclic adenosine 3':5'-monophosphate (cyclic AMP) and corticosterone isolated ratadrenal cells was increased by cholera enterotoxin. Both responses were accompanied by a lag period which is characteristic of other known actions of enterotoxin. The duration of the lag period in the production of corticosterone depended on the concentration of enterotoxin; with the maximally stimulating amounts it was 30-45 min. 2. Maximum rates of cyclic AMP and corticosterone synthesis, after the lag period, were constant for at least 1 h. Although the maximum rate of corticosterone formation was the same as that obtained adrenocorticotropic hormone, the maximum rate of cyclic AMP formation was only 8-10% of that with adrenocorticotropic hormone. 3. Pretreatment of the cells with enterotoxin ahd no effect on their subsequent steroidogenic response to maximally stimulating amounts of adrenocorticotropic hormone. 4. Cycloheximide inhibited the effect of both enterotoxin and adrenocorticotropic hormone on corticosterone production. 5. Enterotoxin stimulation of both cyclic AMP and corticosterone formation was dependent on the presence of Ca2+ in the medium although the Ca2+ requirement was not same as that for adrenocorticotropic hormone. Thus, EGTA at concentrations which completely abolished the effect of adrenocorticotropic hormone caused only a partial reduction in the effects of enterotoxin. 6. Exogenously added choleragenoid and gangliosides abolished the effects of enterotoxin without having any significant effect on the response of the cells to adrenocorticotropic hormone. 7. After treatment with neuraminidase, the adrenal cells showed an increased response to enterotoxin in terms of both cyclic AMP and corticosterone formation which was due to a combination of two effects: (a) increased rate of synthesis of both compounds and (b) shortening of the characteristic lag period. This is in sharp contrast to the results obtained with adrenocorticotropic hormone where neuraminidase-treatment made the cells less sensitive to adrenocorticotropic hormone.


[3H] norepinephrine binding by rat glial cells in culture. Lack of correlation between binding and adenylate cyclase activation. Subcellular fractions prepared from rat glial cells in culture (clonal line c6) were used in an attempt to characterize the adrenergic receptor involved in adenylate cyclase activation. Both [3H] norepinephrine binding and enzyme activation were measured under identical experimental conditions. Binding sites for norepinephrine could be detected; their main characteristics were: apparant Km: 4 - 10-6 M, macimal capacity: 20 pmol/mg protein. Their stereospecificity towards structually related drugs was found to be different from the stereospecificity of the receptor involved in adenylate cyclase activation. Thus, 3-methoxydopamine (a competitive inhibitor of norepinephrine for adenylate cyclase activation), phenylephrine (a partial adrennergic agonist) and the blocking agent propranolol were unable to compete with [3H] norepinephrine for binding. On the other hand, several molecules like dopa bearing a catechol group and which are unable to interact with the adenylate cyclase as agonists or competitive inhibitors strongly inhibited [3H] norepinephrine binding. As in several other systems so far studied, the presence on the glial cell's membrane of a large number of "catechol-binding sites" makes it difficult to characterize the beta-adrenergic receptor.


Transformation of the rat uterine estrogen receptor after partial purification. Warming crude ratuterine cytosol after the addition of [3H] estradiol accelerates the association of the 4-S estrogen-binding protein with a second macromolecule, resulting in the formation of the 5-S estrogen-binding protein. To determine whether the 5-S estrogen-binding protein consists of two similar or dissimilar subunits, uterine cytosol was subjected to a number of fractionation procedures that separate macromolecules by solubility, molecular gel sieving, sedimentation rate, ionic charge, and heat lability. Following each of these methods, the fraction containing the 4-S estrogen-binding protein was incubated at 28 degrees C; each of the these 4-S estrogen-binding protein-containing fractions retained its capacity to completely transform to the 5-S estrogen-binding protein. In samples subjected to partial purification procedures, it was necessary that the buffer contain 40 mM Tris, 60 mM Tris, 60 mM KC1, 1-10 MM dithiothreitol, and 1 M urea at pH 7.4, in order to accomplish the 4-S to 5-S estrogen-binding protein transformation at 25 degrees C. Formation of the 5-S estrogen-binding protein requires association of the 4-Estrogen-binding protein with a molecule identical to or very similar to itself.


Stimulation of cartilage amino acid uptake by growth hormone-dependent factors in serum. Mediation by adenosine 3':5'-monophosphate. The effects of growth hormone-dependent serum factors on amino acid transport and on cartilage cyclic AMP levels in embryonic chicken cartilage were studied in vitro. Cartilages incubated in medium containing rat serum showed a significantly greater uptake of alpha-amino [1-14C] isobutyrate or [1-14C] cycloleucine than control cartilages incubated in medium alone. Normal rat serum (5%) added to the incubation medium also caused an increase in cartilage cyclic AMP content (from as little as 23% to as much as 109%). The factors in serum which increase cartilage cyclic AMP and amino acid uptake are growth hormone dependent, since neither growth hormone itself nor serum from hypophysectomized rats restores these serum factors. Studies comparing the ability of sera with varying amounts of growth hormone-dependent factors to stimulate amino-aminoisobutyrate transport and to increase cartilage cyclic AMP show a striking linear correlation between the two effects (r=0.977). Theophylline and prostaglandin E1, WHICH RAISE CARTILAGE CYCLIC AMP also increase amino-aminoisobutyrate transport. Exogenous cyclic AMP, N6-monobutyryl cyclic AMP and n6, 02'-dibutyryl cyclic AMP increase cartilage amino-aminoisobutyrate transport. The data are compatible with the thesis that growth hormone-dependent serum factors increase cartilage amino acid transport by elevating cartilage cyclic AMP.


[The potential nuclease activity of the T antigen of SV-40 virus]. SV40 T-antigens were isolated from an extract of golden hamster tumours by precipitation with ammonium sulphate with subsequent fractionation on DEAE cellulose. The degree of purification of the preparation proved to be about 100-fold; it, however, contained an admixture of several cell proteins. Treatment of the DNA of the calf thymus with the T-antigen preparation in the presence of magnesium ions decreased the viscosity of the DNA solution during the first hour of incubation. T-antigen inactivated by heating, and also a fraction of normal hamster tissues analogous to it produced no such effect. In case of centrifugation in the saccharose gradient the constant of DNA sedimentation fell after the treatment with T-antigen from 285 to 165, this corresponding to about4--5-fold reduction of molecular weight of the DNA. The data obtained indicated that the partially purified T-antigen preparation possessed endonuclease activity.


Long survival after operation for cancer of the lung. A survey is presented of 293 cases of lung cancer operated on by one surgeon over a period of 35 years. Lobectomy was done in 86 cases with a mortality of 3.5 per cent. Simple pneumonectomy was done in 89 cases with a mortality of 20 per cent, and radical pneumonectomy in 118 cases with a mortality of 15-25 per cent. The survival rate after 5 or more years for lobectomy was 41 per cent (34 patients). After simple pneumonectomy 21 patients (30 per cent) lived 5 years or more, and after radical pneumonectomy 39 patients (39 per cent) lived 5 years or more. The survival of the patients in whom the excised lymph nodes were involved by growth is presented. It is pointed out that at least 1 in 3 patients with cancer of the lung treated by operation was alive 5 years later. The very good long term results are presented including the 10-year and 20-year survival rate. Two patients are still alive after 30 and 34 years respectively. One patient is alive 29 years after a lobectomy and 13 years after a contralateral lobectomy for a new primary cancer. A second new cancer occurred in 9 patients. Continuance of smoking is shown to be harmful. The factors affecting the long term freedom from recurrence are discussed. The cell type of the cancer is shown to be important. Squamous-celled growths have the best results and oat-celled and small-celled the worst, although long term survival can follow operation for both these. The occurrence of a pleural infection after operation may be favourable to long term freedom from recurrence and this way probably so in 2 patients who lived 25 and 27 years respectively and 1 patient who is still alive after 34 years. On the whole the evidence is unconvincing that much benefit may follow radiotherapy combined with operation.


[Temperature dependence of the epr spectra of a spin-labeled myosin and its complex with adp]. The effect of temperature on ESR spectra of spin-labelled myosin and myosin-ADP complex was studied. It was shown that mobility of the sptrongly immobilized spin label almost did not change with temperature (-2 degrees C-37 degrees C) whereas mobility of the slightly immobilized spin label changed in a complicated manner and not monotonously. The observed changes in mobility of the slightly immobilized spin label and characteristics of the ESP spectrum of the myosin-ADP complex suggest that temperature causes continuous changes of the structure in the local environment of the binding sites of the spin label, and on the average, one conformation of myosin and myosin-ADP complex molecules exists at any temperature.


Oestrogen receptors in breast cancer: a changing concept. Oestrogen receptors were found in 156 (73%) out of 214 primary breast cancers and in 98 (58%) out of 168 metastatic deposits. These proportions reached 82% and 70% respectively in the second half of the study.  Receptors were not found in samples of normal breast tissue but small amounts were present in tissue from some hyperplastic lesions and in male gynaecomastia.  Receptor concentrations in the malignant samples were evenly distributed over a wide range of values, suggesting that even "negative" tumours might contain trace amounts undetectable by the method used.  Each tumour was characterized by a given level of receptor concentration.  In most cases the amounts found in the invaded axillary nodes and their corresponding primary tumours were the same.  We suggest that quantitative rather than qualitative assessment should provide an appropriate criterion for studies of biochemical and clinical correlations.


Role of respiratory viruses in childhood mortality. Respiratory viruses have been identified at necropsy in the lungs of 13 out of 24 children who died with observed acute respiratory illness. The histological appearances of the lungs supported the association between virus and death in each of these 13 children and suggested an unidentified virus aetiology in a further five cases. Histological appearances compatible with bacterial infection were found in the lungs of only two of the 24 children. Similar virus and histological findings have been reported in about one-third of victims of the sudden infant death syndrome (cot deaths), indicating a rapid unobserved respiratory virus infection as the most likely mode of death in this group. Evidence that respiratory viruses may be involved in a larger proportion of sudden unexpected deaths, perhaps as antigens in a hypersensitivity reaction, is discussed. Respiratory viruses seem the major identifiable agents contributing to the maintenance of the postneonatal mortality rate since acute respiratory illness and the sudden infant death syndrome together account for about two-thirds of deaths at this age.


Cerebellar responses to teleceptive stimuli in alert monkeys. Discharges of single Purkinje cells in the intermediate and lateral zones of the cerebellar cortex and of neurons in the interpositus and dentate nuclei were recorded in alert monkeys during the presentation of intense auditory and visual stimuli. Concomitant monitoring of the electromyogram (EMG) demonstrated that these stimuli evoked characteristic startle responses in most instances. Firing patterns of cerebellar nuclear cells to auditory stimuli could be categorized into four types, the most common of which consisted of a short-latency acceleration of discharge, followed by a decrease in activity, and in most cells by a later period of facilitation. Simple spike discharge patterns of Purkinje cells consisted largely of prolonged increases or decreases in firing rate, although more complex patterns were seen. In almost 50% of the Purkinje cells tested, complex spikes were evoked by the auditory stimuli. Comparison of simple spike responses of Purkinje cells and of the discharges of cerebellar nuclear cells to auditory and visual inputs revealed that, except for a longer latency, the discharge pattern evoked by flash stimuli was identical to that evoked by sound in all instances. By contrast, in about one-third of the Purkinje cells with related complex spike discharge, complex spikes were evoked by stimuli of only a single modality. Comparison of the times of changes in nuclear and Purkinje cell activity suggests that the initial change in nuclear cell discharge was due to an increase in mossy fiber activity, while the subsequent decrease resulted from Purkinje cell inhibition evoked by mossy and climbing fiber inputs. The absence of increases in nuclear cell discharge at the time of most decreases in Purkinje activity indicates that removal of Purkinje inhibition does not have a major effect on the discharge rates of individual nuclear cells. The data also suggest that excitation of nuclear cells via climbing fiber collaterals played only a minor role in influencing their discharge. Since most EMG changes occurred after or at about the same time as the initial changes in cerebellar discharge, it is unlikely that the initial changes in cerebellar activity were a result of feedback from contracting muscles. It is proposed that the similar discharge patterns of cerebellar neurons to auditory and visual input results from a convergence of these inputs on a structure which projects to the cerebellum as mossy fibers.


Quantitative aspects of the shift-effect in cat retinal ganglion cells. A sudden displacement of a pattern, even when far away from the classical border of a concentrically organized receptive field, elicits a transient excitatory response in on- and off-center retinal ganglion cells (shift-effect). Peak response and latency of the shift-effect have been studied under various stimulus conditions in on- and off-center ganglion cells. Increasing the retinal distance between the site of stimulation and receptive field yields responses of increasing latency corresponding to an intraretinal conduction velocity of about 0.35 m/sec. Beyond a distance of 20 degrees response amplitudes decrease steadily in on- and off-center neurons. Shift amplitude and contrast of the shifting grating have almost no influence above a threshold of 30 min of arc and 10-20% modulation respectively (all-or-none behavior). Shifts of decreasing velocity (600-10 degrees/sec) increase the latency. The response magnitude decreases only at very slow movements. Other properties of the shift-effect are briefly reported. Possible functional significances in relation to eye movements and brightness perception and the retinal pathway of the shift-effect are discussed.


Generation of spike trains in CNS neurons. The membrane potential waveforms to be expected from many asynchronous inputs to CNS neurons are described, along with modes for repetitive firing through which the input waveforms are converted into spike trains. Area beneath a postsynaptic potential (PSP), rather than PSP peak height, is shown to be an important parameter susceptible to modification. Occasional crossings of threshold produce occasional spikes, but a sustained depolarizing waveform which attempts to hold the membrane potential above threshold elicits rhythmic firing. Firing rate is graded with the amount by which the synaptic depolarizing currents exceed the minimum current for rhythmic firing (approximately rheobase). A systematic sequence of alterations in the membrane potential trajectory between spikes, quite different from those of receptors and invertebrate neurons, may control the firing rate and give rise to sudden changes in the "gain" of this conversion of depolarizing current into firing rate. The different implications of synaptic location during the occasional spike mode and the rhythmic firing mode are discussed, as is the role of the antidromic invasion of the soma-dendritic region during rhythmic firing. Less frequently an"extra spike mode" is seen where depolarizing afterpotentials following a spike themselves cross threshold to elicit an extra spike, which may similarly elicit another extra spike, etc., in a regenerative cycle. The character of the underlying depolarizing afterpotentials (or "delayed depolarizations") is reviewed, along with theories for their origin from the antidromic invasion of the dendritic tree. The stereotyped burst firing patterns characteristic of the extra spike mode can also be seen in deafferented neurons and neurons studied in chronic syndromes such as epilepsy and central pain. This raises the question as to whether some disease states may augment extra spike firing, thus multiplying many-fold the response to a normal input.


Modulation of cortical and pyramidal tract induced motor responses by electrical stimulation of the basal ganglia. Two general mechanisms based on anatomical studies are possible for modulation of motor activity by the caudate nucleus and globus pallidus. These mechanisms are: (1) modulation of the output of cortical neurons that exert motor influences; and (2) modulation of subcortical neurons that exert motor influences. Differentiation between these two mechanisms was accomplished in the present study by two experimental approaches, both of which employed the conditioning-test paradigm. The first approach was an investigation of caudate nucleus or globus pallidus modulation (conditioning stimulus) of flexor responses of the anterior tibialis muscle elicited by electrical stimulation of the sensorimotor cortex (test stimulus) or pyramidal tract (test stimulus). These investigations were carried out in the intact and in decorticate cats. The second approach was an analysis of modulation or cortically induced pyramidal tract responses (direct and indirect, D-I potentials) by conditioning shock trains delivered to various loci within the caudate nucleus or globus pallidus. Both approaches were designed to determine whixh inhibitory and facilitatory motor influences of the basal ganglia occurred at a cortical or subcortical level. Simultaneous stimulation of a locus within the caudate nucleus and the sensorimotor cortex evoked either an enhancement, reduction or no alteration of the cortically induced increase in flexor responses (measured  by Ia afferent activity, EMG, myogram). In contrast, no inhibitory influences occurred from caudate nucleus stimulation upon pyramidal tract induced flexor responses in either the intact or decorticate preparation. Inhibitory loci were distributed toward the rostral portion of the caudate nucleus, whereas facilitatory loci were distributed throughout; this distribution was statistically significant (chi2; P less than 0.01). Only enhancement or no influence upon cortical induced or pyramidal tract induced responses were obtained by conditioning stimuli to the globus pallidus. In the unanesthetized but immobilized cat, trains of shocks delivered to the caudate nucleus enhanced, reduced or had no influence upon the cortically evoked direct (D) and indirect (I) potentials recorded in the bulbar pyramidal tract. The distribution of facilitatory and inhibitory loci was organized in a similar fashion as in theanesthetized preparation. From these observations, a model was proposed in which the output of the caudate nucleus exerts both facilitatory or inhibitory modulation of the tonically active globus pallidus cells. The latter in turn predominantly or exclusively facilitate output of pyramidal tract neurons as well as the output of subcortical structures; both effects facilitate motor responses at the spinal level.


The use of frozen cells in the microtitre serum neutralization test for infectious bovine rhinotracheitis. Some parameters of the microtitre serum neutralization test were examined when using bovine fetal kidney cells derived from stocks stored in liquid nitrogen. In replicate tests with one serum there was no significant difference (p greater than 0.05) between titres calculated on the basis of cytopathic effect when either two or four wells were used per serum dilution. Also, there was no significant difference between titres calculated on the basis of a cell staining method when either two or four wells were used per serum dilution. When titres obtained by the 2-well-cytopathic effect method were compared with those obtained by the 2-well-stain method differences were not significantly different and it was concluded that the latter method using frozen cells constituted a practical and reproducible test. No significant titre variation occurred in cells serially passaged after having been frozen, or in cells derived from five different fetuses. Titres of sera from 41 cattle conformed to a log normal distribution pattern.


Infectious bovine keratoconjunctivitis I. Experimental production. One or both eyes of 20 calves were inoculated one or more time with variou(s combinations of microorganism (live oor killed Moraxella bovis, infectious bovine rhinotracheitis virus, bovine adenovirus, bovine parainfluenza-3 virus and Mycoplasma bovoculi) by conjunctival instillation or direct inoculation of the conjunctivea or cornea. The eyes of all the calves received natural or artificial ultraviolet irradiation. Neither the adenovirus nor parainfluenza-3 virus became established in the eye or produced keratoconjunctivitis. Both M. bovis and infectious bovine rhinotracheitis virus became established in the bovine eye and produced disease. Subconjunctival or intracorneal inoculation of M. bovis caused a severe disease, simulating natural infectious bovine keratoconjunctivitis. Only the intracorneal inoculation of mycoplasma produced severe keratoconjunctivits. Eyes that on initial exposure to M. bovis became severly inflamed were more resistant to a second or third exposure to M. bovis, presumably by enhanced local defence mechanisms.


Further observations on the characteristics of a bovine parainfluenza-3 variant. Calves were inoculated with a bovine para-influenza-3 variant to determine its pathogenicity and the stability of its cytopathic feature and its inability to agglutinate chicken erythrocytes. The inoculated calves and one contact animal developed an immune response without significant clinical illness. The clinical response in calves was similar to that induced by the parent virus strain. The variant was shown to retain its characteristic cytopathic effect for Madin Darby bovine kidney cells and its property of hemagglutination following one passage in the natural host.


Utilization of triaryl phosphates by a mixed bacterial population. A mixed bacterial population that has been isolated by enrichment culture is capable of growth on Fyrquel 220, a commercial triaryl phosphate lubricant, as sole carbon source. The mixture was dominated by a yellow, Gram-negative rod which made up greater than 60% of the mixture. However, all attempts to grow this organism in pure culture on triaryl phosphate were unsuccessful. The mixed population was also capable of growth on tri-o-cresyl phosphate, trixylenyl phosphate, and triphenyl phosphate as sole carbon sources. Viable cell numbers increased 20- to 30-fold, reaching a maximum after 72-96 h growth. Only a small portion of the triaryl phosphate was used for growth; the major part was emulsified and remained in the culture medium. No evidence of extracellular enzymes capable of triaryl phosphate degradation could be found in concentrates of the culture supernatant after growth, though traces of what may have been triaryl phosphate breakdown products were observed. Cell-free extracts of the mixed culture catalyzed the release of inorganic phosphate when incubated with Fyrquel 220, tri-o-cresyl phosphate, trixylenyl phosphate, or triphenyl phosphate, indicating the presence of a phosphotriesterase or of a phosphodiesterase of wide specificity.


Composition of the capsular polysaccharides of Clostridium perfringens as a basis for their classification by chemotypes. An analytical procedure, using gas-liquid chromatography, was developed for the identification of the per(trimethylsilyl) ethers of the constituent monosaccharides obtained from the capsular polysaccharides of Clostridium perfringens Hobbs 5, Hobbs 9, Hobbs 10, and NCTC 10578. Qualitative and quantitative differences between the major polysaccharide components enabled the differentiation of the four strains of C. perfringens investigated.


Conversion of spheroplast symbiotes in a leafhopper, Helochara communis Fitch (Cicadellidae: Homoptera). In mycetomes of leafhoppers, Helochara communis, ultrastructural and histochemical studies revealed that spheroplast symbiotes (t-symbiotes) were converted to 'a-symbiotes' (so-called), with apparent loss of DNA--a phenomenon reminiscent of rhizobium-bacteroid conversion in certain legume nodules. Additional t-symbiotes were incorporated into the substance of these 'a-symbiotes'.


Progeny resulting from complementation between mutants of vesicular stomatitis virus. The complementation properties of the virus progeny released from cells mixedly infected with mutants of vesicular stomatitis virus belonging to four different complementation groups have been examined. The group IV mutant, tsW16B, was tested in combinations with three group I mutants (tsW4, tsW28, and tsG11), one group II mutant (tsG22), and one group III mutant (tsW29). Virus stocks were grown from isolated plaques appearing on the cell monolayers used to assay the mixed infection yields and tested, in a second series of mixed infections, for their ability to complement each of the two parents. It was found that the virus harvested from each one of the first series of mixed infections contained mutants of both parental types.


Cerebellar capillary hemangioblastoma: its histogenesis studied by organ culture and electron microscopy. A capillary hemangioblastoma from the vermis of a 21-year-old man with von Hippel-Lindau's disease was successfully maintained in organ culture systems on Millipore filter platforms and gelatin sponge foam matrices up to 48 days. Three cell types--endothelial cells, pericytes, and stromal cells--identified by their fine structural features and their architectural relationships to vascular lumens and to the extracellular space were recognized in the original tumor. By light microscopy the explants showed, in the late stages, increased lipid droplets in the stromal cells and perivascular hyaline thickening. By electron microscopy, endothelial cells, pericytes, and stromal cells remained distinguishable as cell types. However, the stromal cells demonstrated, in addition to an increase of their normal complement of microfilaments and lipid droplets, features that were more characteristic of the two other cell types, i.e. micropinocytotic vesicles and the formation of zonula occludens junctions and hemidesmosomes. Basement membranes also became more apparent around the stromal cells. With increasing time in vitro, there was a striking increase in mature collagen fibers in the extracellular space. The roles of the different cell types in capillary hemangioblastoma and the histogenesis of the stromal cells are discussed in light of these observations. It is concluded that the capillary hemangioblastoma consists of multiple cell lines--endothelial cells, pericytes, stromal cells and, occasionally, hematopoietic cells--all of which are neoplastic and replicate in parallel with one another. Stromal cells may be regarded as an aberrant monopotential cell type which shares with the endothelial cell and pericyte a common mesenchymal, presumably angiogenic, ancestry, and may, on occasion, display morphological features, such as increased basement membrane formation and the formation of zonula occludens junctions, which recall its angiogenic lineage. However, interconvertibility between endothelial cells and stromal cells does not appear to occur in vivo or in vitro.


An assessment of the World Health Organization classification of the histologic typing of lung tumors applied to biopsy and resected material. In a study of the World Health Organization classification of the histologic typing of lung tumors, sections from a total of 740 patients in the Medical Research Council Study of Cytotoxic Chemotherapy, 182 of whom also had positive preoperativebronchial biopsies and 231 involved lymph nodes in the resected specimens, and from 30 patients in the Medical Research Council Trial of Surgery and Radiotherapy in Small or Oat-celled Carcinoma of the Bronchus have been assessed. Of the 740 primary tumors from the Study of Cytotoxic Chemotherapy, 71% were placed in Type I, 12% in Type II, 9% in Type III, and 7% in Type IV. Only 2 primary tumors could not be typed. A blind comparison of the type of primary tumor and bronchial biopsy showed that the biopsy was a good indicator of the type of the primary tumor. A bind comparison of the primary tumor and involved lymph node also showed a close degree of agreement. However, when the type of the primary tumor was assessed in the presence of the involved node. Tumors were placed in Type IV far less frequently than when assessed blind. It is concluded that the World Health Organization classification is applicable to primary tumor, bronchial biopsy, and involved node, that the biopsy is a valuable indicator of the type of the primary tumor, and that the apparent type of the involved node should not be allowed to over-influence the pathologist in deciding on the type of the primary tumor when both are assessed together.


The glycolytic enzyme activity of the human cervix uteri. The activity of the following enzymes was studied in normal, precancerous, and malignant biopsies from the human cervix uteri: hexokinase (HK), phosphofructokinase (PFK), pyruvate-kinase (PK), lactate dehydrogenase (LDH), and glucose-6-phosphate dehydrogenase (G-6-PDH). In precancerous conditions, i.e., dysplasia and carcinoma in situ without any signs of invasive carcinoma, only PK showed moderate but significant activity increases. A rise in enzyme activity in biopsies histologically classified as carcinoma in situ was found to signal the presence of invasive carcinoma in other parts of the cervix. In invasive carcinomas of the cervix, all the enzymes studied showed a two- to four-fold increase (p less than 0.01) as compared to the normal cervix. The present study failed to reveal significant differences between enzyme activities in biopsies from patients in Stage I, II, and III; no correlation could be established between enzyme activity and prognosis.


Cervical synovial sarcoma at the bifurcation of the carotid artery. Cervical synovial sarcoma has been reported in only 15 cases; in none of these was there any association with a vascular structure. A patient with cervical synovial sarcoma encompassing the bifurcation of the left carotid artery underwent a left radical neck dissection, including excision of the left external carotid artery and the hypoglossal nerve. The tumor was dissected away from the common carotid and internal carotid arteries. Although the fibrous pseudocapsule of the tumor had not involved the fibrous adventitia of the carotid arteries. Radiation therapy with 6000 rads tumor dose was administered to a wide local field. There is no evidence of recurrence 1 year postoperatively. Histologic and ultrastructural findings were similar to those reported previously in a case of synovial sarcoma of an extremity.


Salivary gland neoplasms following atomic radiation: additional cases and reanalysis of combined data in a fixed population, 1957-1970. A reappraisal was made of the relationship between exposure to atomic radiation in 1945 and the occurrence of salivary gland tumors in heavily exposed survivors. This was made possible by the discovery of 8 additional patients; the total now reported consists of 30 cases. In those exposed to 300+ rads, the number of cases was significantly (p less than .01) greater than expected. This was found for the combined group and for malignant types, but not for histologically benign tumors. Our prior conclusion regarding the high-dose radiation effect upon salivary gland neoplasia remains unchanged in the reanalysis; however, our suggestion that younger persons were at significantly greater risk is weakened.


On the mechanism of hormone action in 7,12 dimethylbenz(a)anthracene-induced mammary tumor. I. Prolactin and progesterone effects on estrogen receptor in vitro. The presence of ER in DMBA-tumors was demonstrated by the use of dextran-charcoal assay, sephadex chromatography, sucrose gradient sedimentation, and organ culture techniques. It was found that tumors have binding sites ranging from 10-13 to 10-15 moles/mg protein, and a dissociation constant of ER 10-9 to 10-10 M. In experiments with tumor explants, prolactin-insulin significantly stimulated ER binding capacity, as compared with control without prolactin. This stimulation was tissue-specific and inhibited by progesterone. Insulin had a synergistic effect on prolactin stimulation of ER. Our results presents a plausible explanation for tumor responses to these hormones in vivo. This interaction of prolactin, estrogen, and progesterone may be a common phenomenon for all estrogen-responsive tissues.


Thymus-dependent lymphocyte levels in bronchogenic carcinoma: correlations with histology, clinical stage, and clinical course after surgical treatment. The in vitro spontaneous lymphocyte rosette (T cell) assay was used to determine cellular immunologic competence in 112 patients with bronchogenic carcinoma. Among preoperative patients with localized tumors. T cell levels were significantly lower than in 237 normal controls. With advanced stages of disease, T cell levels declined progressively among patients with squamous cell carcinoma, oat cell carcinoma, and undifferentiated carcinoma, but not among patients with adenocarcinoma. Squamous carcinoma patients considered cured had persisting low T cell levels, but cured adenocarcinoma patients had normal levels. Serial determinations that showed a fall in T cell levels preceded the development of clinically evident metastases by an average of 2.5 months. Postoperative patients with rising T cell levels have remained clinically free of disease. The data indicate that T cell levels correlate with extent of tumor and clinical course of patients with bronchogenic carcinoma. The assay may, therefore, provide a rational basis for the selection of patients who are at high risk for the development of recurrence after surgical resection and who may benefit from the early institution of adjunctive therapy.


"Residual" mammary carcinoma following simulated partial mastectomy. Treatment of mammary carcinoma by partial mastectomy rather than by total mastectomy and axillary dissection may diminish the chances of long-term cure by risking incomplete removal of all local carcinoma at the initial operation. This study was undertaken to determine by pathologic examination how often carcinoma might remain in the breast and axilla after partial mastectomy. The operation was simulated in 203 mastectomy specimens after operations for unilateral invasive carcinoma. In so far as could be determined on gross examination, the entire primary lesion was included in the quadrant which was excised in the simulated procedure. Among 100 women with primary lesions less than 2 cm in diameter, 26% had carcinoma in the breast which remained after simulated partial mastectomy. Six percent of them also had axillary node metastases. An additional 30% only had axillary node metastases. When the primary lesion was more than 2 cm in diameter, 38% of patients had carcinoma in the breast after simulated partial mastectomy, of whom 29% also had axillary metastases. After simulated partial mastectomy, carcinoma was found in 80% of breasts from patients with lesions in the subareolar area, in contrast with 25-35% of patients with a primary carcinoma in one of the four quadrants. None of the 9 patients with medullary and colloid carcinomas that measured under 2 cm had axillary metastases or carcinoma in the breast outside of the primary quadrant. The findings suggested that a familial history of breast carcinoma or a large primary lesion may be associated more often with multifocal disease, but factors such as age at diagnosis, axillary status, and the mammogram report did not have significant predictive value for distinguishing between patients who did or did not have carcinoma in breast tissue after the primary had been removed by a simulated partial mastectomy.


The genetics of childhood cancer.

The genetics of childhood cancer. Retinoblastoma should not be considered as the exceptional childhood cancer that shows dominant inheritance, but rather as the typical childhood cancer that embraces a prezygotic and postzygotic subgroup. Postzygotic cases are conceived as involving mutation as a first step too, but with the mutation in somatic rather than germinal cells. In both prezygotic and postzygotic cases a second event, possibly mutational, occurs before the cancer is initiated. The embryonal cancers are all visualized as genetic disease, and their frequencies limited by gene mutability. This mutability can be increased by environmental agents. Prezygotic cases may develop other primary tumors in other tissues. They may also have affected family members, the probability of affected offspring approaching 50%. A diagnostic test to identify prezygotic cases among those with a single primary tumor and a negative family history is sorely needed.


The evaluation of abdominal masses in children with emphasis on noninvasive methods. A roentgenographic approach. Intravenous urography with total body opacification, and tomography as required, often give the most information toward evaluating abdominal masses in children. Ultrasonography is a noninvasive procedure which defines normal structures and differentiates cystic and solid tumors. The combination of these studies gives sufficient information about renal tumors to plan for possible surgery. Arteriography is not necessary for the diagnosis of Wilms' tumor, nor its surgical or medical management. Pseudotumor of the kidney is due to focal cortical hyperplasia. It can be diagnosed by nephrotomography, renal arteriography or renal scanning. The latter method is most accurate and has the lowest morbidity. Aortography is advisable in the evaluation of a patient with pheochromocytoma in an attempt to locate multiple tumors. Determining the extent of abdominal neuroblastoma by angiography and lymphangiography does not appear to influence the mode of therapy, not the survival rate; therefore, invasive diagnostic procedures do not appear to be indicated in neuroblastoma. Angiography is necessary in the evaluation of liver cancer. If one lobe is determined to be free of disease, lobectomy is a possible cure. Splenic cysts and choledochal cysts can be diagnosed by noninvasive methods such as ultrasonography or radioisotope scanning. Arteriography and percutaneous opacification are not necessary to make these diagnoses.


Advances in the treatment of Wilms' tumor. The addition of chemotherapy to older standard treatment with surgery and radiotherapy has increased survival to more than 80% in localized Wilms' tumor, and about 50% in metastatic disease. Actinomycin-D and vincristine have proved to be the two most active agents. Multiple-course maintenance treatment with actinomycin-D has allowed fewer relapses, but not a significantly different survival rate than a single course of this drug. Use of actinomycin-D and vincristine in combination has been under study; initial indications show that the combination is superior to either drug employed singly. The need for postoperative radiotherapy in totally resected tumors confined to the kidney is also under study. Adriamycin is at present the most encouraging new drug that has had clinical trial.


Radiation therapy of tumors of the central nervous system in childhood. The results of radiation therapy for CNS tumors of childhood are presented and discussed according to histologic type. In this material the 5-year recurrence-free survival rates were about 35% for medulloblastomas, 48% for the hypothalamic and brain stem tumors, 86% for lowgrade ependymomas, and 57% for germinomas. Complications or CNS injury from irradiation were rare and the quality of life in survivors was generally good. Current methods of radiation therapy may possibly improve these results. Tumors of the CNS in children do not necessarily carry a hopeless prognosis; aggressive therapy is indicated.


Dissociations between changes in myocardial cyclic adenosine monophosphate and contractility. The relationship between changes in the myocardial concentration of adenosine 3':5'-cyclic monophosphate (cyclic AMP) and cardiac contractility was studied in guinea pig and rat myocardium. When isolated perfused guinea pig heart were perfused with 10-5-M papaverine, a potent inhibitor of cyclic AMP phosphodiesterase activity, myocardial cyclic AMP concentration increased significantly from 1.7 plus and minus 0.2 (SE) pmoles/mg protein (N equal 12) to 3.3 plus and minus 0.2 pmoles/mg protein (N equal 12), and the percent of phosphorylase aual 6) (P less than 0.01). However, perfusion with papaverine had no effect on contractility in the absence or the presence of exogenous epinephrine. In perfused rat hearts, 10-5 M glucagon increased myocardial cyclic AMP concentration from 1.5 plus and minus 0.1 pmoles/mg protein (N equal 12) to 2.6 plus and minus 0.1 pmoles/mg protein (N equal 12) (P less than 0.001). In contrast, cyclic AMP levels did not increase detectably in guinea pig heart perfused with glucagon. Glucagon increased adenylate cyclase activity more than twofold in rat myocardial broken cell preparations but failed to stimulate the enzyme in preparations from guinea pigs. Despite these differences, the positive inotropic effects of glucagon on rat and guinea pig hearts were very similar over a wide dose range. Thus, with both papaverine and glucagon, changes in cardiac contractility were dissociated from stimulation of adenylate cyclase activity, increases in myocardial cyclic AMP levels, and conversion of phosphorylase b to phosphorylase a in perfused hearts.