Light and electron microscopic features of a pituitary adenoma in Nelson's syndrome.

Electron microscopy of an amphophil pituitary adenoma surgically removed from a 51-year-old woman who had Nelson's syndrome revealed that the tumor was composed of melanocorticotroph cells. This finding is consistent with the view that in the human pituitary gland one single cell type produces both adrenocorticotropic hormone (ACTH) and melanocyte-stimulating hormone (MSH). In contrast to the ultrastructure of pituitary adenomas associated with Cushing's syndrome, no or only very few microfilaments were detected in the cytoplasm of the tumor cells, suggesting that adrenocortical steroids are required for the formation of microfilaments. The presence or absence of microfilaments in the tumor cells may be regarded as a distinguishing ultrastructural feature between Cushing's syndrome and Nelson's syndrome. It appears that changes in the level of circulating corticoids may affect the ultrastructural features of melanocorticotroph cells not only in normal pituitaries but also in adenohypophyseal adenomas.


Steroid-binding proteins in carcinoma of the human male breast.

Receptor sites for a group of four steroid hormones were estimated by competitive binding assay of a biopsy specimen of carcinoma from a male breast. The results indicate that the tissue contained a significant number of binding sites for 17beta estradiol, and probably for dihydrotestosterone and progesterone, but not for testosterone. These preliminary data suggest that, as in the female, certain mammary carcinomas in the male may be under the influence of steroid hormones. Correlation of such data with responses to therapy may help to establish a more rational approach to the treatment of carcinoma of the male breast.


Positive mono-spot test in histiocytic medullary reticulosis.

A case of rapidly fatal histiocytic medullary reticulosis in which two positive mono-spot tests were found is presented. In addition, there was a low liter antibody to EB virus. The patient died with a bleeding diathesis terminally. Postmortem examinations confirmed the diagnosis of histiocytic medullary reticulosis.


Isolation of mycobacteria from clinical specimens by use of selective 7H11 medium.

7H11 agar containing carbenicillin, amphotericin B, polymyxin B and trimethoprim lactate (selective 7H11 or S7H11) was used for the selective isolation of mycobacteria from clinical specimens. This medium as previously described contained 100 mug. carbenicillin per ml. It was found that reducing the concentration of carbenicillin to 50 mug. per ml. made S7H11 less inhibitory to certain strains of mycobacteria. However, some strains of M. kansaii, M. intracellulare, and M. gordonae still do not grow well on S7H11. Of 3,134 clinical specimens (mostly sputum) received, processed, and plated on 7H11 agar and the S7H11 medium, 508 positive specimens were isolated. Of these, 402 were positive on both types of media, 30 were positive on 7H11 only, 19 were positive on S7H11 only, 52 were contaminated on 7H11 but positive on S7H11, while only 5 were contaminated on S7H11 and positive on the plain medium. Thus, the total positive specimens on 7H11 was 437 and total positive specimens on the selective medium was 473. Used in conjunction with nonselective media, S7H11 agar appears to be a valuable culture medium for used in diagnostic mycobacteriology laboratories.


Is a rhinovirus vaccine possible?

Renewal of support for efforts to develop a rhinovirus vaccine seems justified in the light of newer epidemiologic and immunologic studies. The major contribution of RV to acute upper respiratory disease in all age groups but especially in young children emphasizes the public health importance of an effective vaccine. Epidemiologic surveillance of RV infections in widely separated areas has identified two relevant phenomena. First, in each area, certain serotypes were more frequently encountered and tended to persist. Such "common" serotypes accounted for a disproportionate share of the infections recognized and, hence, constitute special targets for immunization. Second, a clear increase over time in the proportion of RV isolates representing higher numbered (types 56-89) serotypes or untypable strains (potentially new serotypes) suggests that new serotypes continue to emerge as the result of progressive antigenic shift. The common origin of the multitudinous RV serotypes so suggested is consistent with the extensive antigenic cross-relations which are becoming evident. Systematic cross-testing with monospecific antisera, especially when high titer sera are employed, has revealed an appreciable number of one-way and reciprocal relations. Largely fortuitous observations of naturally or experimentally infected humans have revealed many additional cross-relations manifested by concurrent response to heterologous RV and presumably attributable to sensitizations resulting from prior RV infections. A model for this has been provided by rabbits immunized sequentially with different potent RV immunogens. Available information as displayed in figure 2 indicates that extensive cross-relations do exist and that sizeable groups of closely related serotypes may be identified. More intensive search for heterotypic response to infection of man coupled with selective use of the rabbit model should define the full extent and strength of cross-relations and identify completely the more closely related groups of serotypes as the basis for formulation of a broadly effective RV vaccine containing a limited number of serotypes. The cross-relations described are based entirely on development of serum neutralizing antibody, the presence of which in man is clearly correlated with relative protection against infection and disease. While this protection may well prove to be mediated largely by concomitant nasal secretory antibody, it is not unreasonable to expect that the antigenic cross-relations also would be manifested in secretory antibody response. These important questions concerning secretory antibody can be best explored in a limited series of volunteer trials with selected cross-related RV serotypes in which homotypic and heterotypic protection could be correlated with serum and nasal secretory antibody. The state of current knowledge, as I view it, is sufficient to justify initiation of such trials at any time.


Neurohormonal control of gonadotropic secretion in primates.

With immunohistochemical techniques, products from both the parvicellular system (such as Gn-RH) and from the magnocellular system (vasopressin, oxytocin, and neurophysins) have been shown to be present in the neurons ending around the hypothalamopituitary portal capillary system. A neurosurgical technique for prolonged collection of pituitary stalk blood has been developed. Gn-RH, vasopressin, and neurophysins have been found in large amounts in pituitary stalk blood. Gn-RH levels were found to vary in a pulsatile fashion, thereby providing direct evidence for hypothalamic modulation of the discharge of LH by the pituitary. The finding of large amounts of estrogen-neurophysin in portal vein blood adds further evidence to the hypothesis that "estrogen" neurophysin may play a role in the control by the hypothalamus over the secretion of gonadotropins.


Case report. Clostridium perfringens septicemia with detection of phospholipase C activity in the serum.

Clostridium perfringens septicemia with massive hemolysis is described in an elderly woman with refractory anemia. This case is highly unusual in that (1) the diagnosis was made during life by discovery of the organisms on a routine peripheral blood film, and that (2) phospholipase C activity was detected in the serum.


Tight-skin, a new mutation of the mouse causing excessive growth of connective tissue and skeleton.

A new dominant mutation, tight-skin (Tsk), is located on Chromosome 2, two recombination units distal to pallid (pa). Heterozygotes (Tsk/+) have tight skins with marked hyperplasia of the subcutaneous loose connective tissues, increased growth of cartilage and bone, and small tendons with hyperplasia of the tendon sheaths. In the loose connective tissue there are large accumulations of microfibrils in the intercellular space. In spite of the increased skeletal size, body weight is not increased. Increase in size of the thoracic skeleton is especially pronounced and leads to pathologic distentsion of the hollow thoracic viscera. Concentration of growth hormone in the pituitary and plasma is normal. Homozygotes (Tsk/TSK) die in utero at 7 to 8 days of gestation. We propose the hypothesis that Tsk might act by causing defective cell receptors with high affinity for a somatomedin-like factor promoting growth of cartilage, bone, and connective tissue and low affinity for a multiplication-stimulating factor promoting embryonic growth.


Ca-activated ATPase activity in subcellular fractions of mouse pancreatic islets.

Ca-stimulated ATPase activity has been demonstrated in homogenates of mouse pancreatic islets. On subcellular fractionation Ca-ATPase activity was found in secretory granules, mitochondria, and microsomes, but not in the postmicrosomal fractions. Highest specific activity was found in the granules. In all active subcellular fractions two Km(Ca) values for Ca-ATPase around 7.0 X 10(-6) and 1.8 X 10(-7) M were estimated. Assuming an ATP hydrolysis:Ca pumping ratio of 1:2, the highest capacity for active Ca transport was found in secretory granules and mitochondria. Concentrations of 40 mM or higher of Na and 10(-5) M cyclic AMP inhibited Ca-ATPase in all subfractions. Caffeine at a concentration of 10 mM inhibited Ca-ATPase significantly in secretory granules and microsomes. Also MG-ATPase activity was demonstrated in the various subfractions. This activity was compared with that of Ca-ATPase at identical concentrations of free metal ions and in the absence or presence of various inhibitors. It was concluded that high-affinity Ca-ATPase and Mg-ATPase are two different enzymic entities. Ca-ATPase may tentatively be assumed to participate in active transport of Ca between intracellular compartments and to constitute a Ca-accumulating system which returns the cytosolic free Ca concentration to the resting state after stimulation of the beta-cells by secretagogues. This enzyme may therefore play a significant role in regulation of insulin release.


The pathogenesis of vesicular stomatitis virus, serotype Indiana, in Aedes aegypti mosquitoes. I. Intrathoracic injection.

Analysis of infectious virus particles after intrathoracic injection revealed that Aedes aegypti mosquito tissues are capable of supporting the growth of vesicular stomatitis virus (VSV), serotype Indiana. Although all tissues assayed (salivary gland, midgut, diverticulum, malphigian tubules, and ovary) were capable of supporting VSV growth, the salivary gland was the only organ capable of maintaining an appreciable amount of virus for periods longer than 9 days postinfection. Electron microscopic studies of infected tissues showed virus particles consistently within the cell cytoplasm of all organs with no evidence of nuclear involvement. Direct evidence of crystalline formation of VSV in the apical cavities of salivary gland tissue was demonstrated.


Operative cholangiography in uncomplicated biliary tract surgery. Review of 354 cholangiography studies in patients without indication of common duct pathology.

The literature of the past ten years is replete with articles on "routine* cholangiography, with particular reference to the overlooked or "unsuspected" stone. Many series do not differentiate between T-tube cholangiograms performed after common duct exploration and those performed on patients with and without the usual clinical or operative indications for common duct exploration. We present a pure operative  cholangiographic study of 354 patients with apparent uncomplicated bilary tract disease; a significant incidence of unsuspected pathologic features are demonstrated. Can a convenient and innocuous procedure like routine operative cholangiography continue to be ignored, even in the most routine of cases?


Selective hepatic venography in primary liver cell carcinoma.

With apparent inoperable hepatic cell carcinoma, knowledge of adequate venous outflow from tumor-free liver tissue permits extensive resection with long survival.


The value of arteriography in the diagnosis and treatment of insulinomas.

Six arteriograms were performed in five patients with insulinoma. The method showed with precision the size and location of the tumor in five instances, whereas in one instance the method was unsuccessful. Arteriography was performed again with more refinements in technic, but was adequate in locating only the largest tumor. Since diagnostic accuracy of arteriography depends on the size of the tumor, and nonpalpable tumors are only occasionally located, the main problems in surgical treatment of insulinomas are unresolved.


Pancreatic tumor of mesenchymal origin presenting as obstructive jaundice.

A case of an unusual tumor occurring in the head of the pancreas is presented. Definitive diagnosis could not be made, the lesion being either a low grade myxofibrosarcoma or a benign fibrous histiocytoma. The literature is discussed and several cases believed to be pancreatic sarcomata are presented, only one of which was definitely of pancreatic origin. No other case of fibrous histiocytoma of the pancreas has been reported.


A quantitative electron microscopic study of atypical structures in normal human cerebral cortex.

Biopsy samples of the cerebral cortex from four normal human brains were examined in the electron microscope for the presence of abnormalities related to neurones and neuroglia. Atypical forms of axons, axon terminals and dendrites, many of them similar to those described in a variety of pathological and experimental material, were found to occur in small but appreciable numbers. Neuroglia exhibiting atypical inclusions occurred but were much less common. Physiologically altered neuronal perikarya were not encountered apart from one neuronal death. An area of 3.6 x 10(5) mu2 was scanned from each brain and atypical structures were categorised and counted. The possible implications of the presence of these structures in normal brain in discussed and the need for neuropathologists and neuroanatomists to be aware of the existence of atypical forms of neuronal processes in normal human and animal brains is emphasised.


Pathogenicity of an attenuated strain of transmissible gastroenteritis virus for newborn pigs.

The pathogenicity of a cell culture-attenuated strain of transmissible gastroenteritis virus for newborn pigs was investigated. Newborn (1- to 2-day-old) pigs were orally given 2 x 10(6) plaque-forming units of attenuated virus. All pigs developed mild diarrhea, but deaths did not occur. As determined by immunofluorescence and villous atropy, infection of the small intestine was limited to the caudal 50 to 66%. Fluorescing cells and atrophic villi were seen from 2 to 3 days until 6 to 7 days after exposure. Attenuated virus-exposed pigs produced circulating virus-neutralizing antibodies detectable as early as 5 days after exposure. By contrast, all pigs orally given 1 x 10(2) pig infective doses of virulent transmissible gastroenteritis virus developed severe diarrhea, and almost all of those not killed died within 2 to 5 days after exposure. In the latter pigs, the entire length of the small intestine, except for the first 4 to 5 cm, was infected with virus by 24 to 36 hours after exposure.


Effects of crude extracts of various plants on infectious bovine rhinotracheitis virus-plaque production.

Extracts of 28 plants were tested without demonstable antiviral activity in an agar-overlay plaque-reduction antiviral assay system, using infectious bovine rhinotracheitis virus and bovine endocardial cell cultures. Ethanolic extract of Narcissus tazetta L bulb elicited antiviral activity by inhibition of viral plaque formation. Antiviral activity was demonstrated against infectious bovine rhinotracheitis and equine rhinopneumonitis viruses. Narcissus tazetta L bulb did not directly inactivate the virus extracellularly. The extract exhibited only limited toxicity to rapidly multiplying bovine endocardial cells at plaque-inhibitory levels and was not cytoxic to preformed confluent cell monolayers. Narcissus extract did not induce the formation of drug-resistant viral strains.


Effects of vaccination infectious bovine rhinotracheitis and simultaneous administration of levamisole on primary humoral responses in calves.

In a study on the primary humoral response of calves vaccinated against infectious bovine rhinotracheitis (IBR) and simultaneously given levamisole, mild but consistent suppression of the group's geometric mean serum-neutralization titer to IBR virus occurred between 12 and 59 days later. The quantitative determination of circulating immunoglobulins (Ig) over the same period indicated a slight decrease in the IgG concentrations from an initial geometric mean concentration of 18.28 mg/ml before calves were vaccinated to 15.29 mg/ml after vaccination, and the control calves (vaccinated and given saline solution only) maintained their prevaccination IgG geometric mean concentration of 20.92 mg/ml. Difference was not observed in the circulating IgM values of the 2 groups. Levamisole had no apparent effect on the circulating Ig-bearing lymphocyte values when compared with control calves during the 24 hours of treatment. It was concluded that a single treatment of levamisole may mildly suppress the primary humoral response to IBR vaccination in calves, but it is unlikely that this would affect the animal's capability to recover from, or maintain resistance to, IBR infection.


The interaction between d-tubocurarine, pancuronium, polymyxin B, and neostigmine on neuromuscular function.

The interaction between pancuronium, d-tubocurarine, polymyxin B, and neostigmine was studied in the rat diaphragm-phrenic nerve preparation. Polymyxin B (5 mug/ml) did not affect twitch tension alone but decreased the pancuronium ED50 from 0.8 mug/ml to 0.32 mug/ml and the d-tubocurarine ED50 from 0.25 mug/ml to 0.15 mug/ml. Neostigmine (0.2 to 10 mug/ml) antagonized pancuronium or d-tubocurarine-induced depression of twitch tension. In contrast, neostigmine (0.001 to 0.2 mug/ml) augmented polymyxin B depression. Similarly, neostigmine (0.1 mug/ml) augmented combined polymyxin B-pancuronium or polymyxin B-d-tubocurarine depression of twitch tension. The authors conclude that polymyxin B potentiates the neuromuscular blockade from pancuronium or d-tubocurarine and that neostigmine further augments this block.


Treating perforated jejunal ulcer, a complication in the Zollinger-Ellison syndrome.

What is considered to represent the fourth case of perforated jejunal ulcer in association with the Zollinger-Ellison syndrome reported in the literature is discussed. Although documented in the original paper by Zollinger and Ellison, this remains a notably rare complication. Generalized peritonitis and related pathophysiological phenomena make initial definitive correction of the ulcer diathesis an extremely riskly undertaking and ill-advised. We submit that primary ulcer resection followed by total gastrectomy when the patient becomes a more suitable operative risk is the preferred method of management. The literature would appear to support this clinical view.


Infiltrating ductal carcinoma of the breast in the postpubertal adolescent: a case report.

Infiltrating ductal carcinoma is an extremely rare neoplasm in the teenage or pre-teenage child. Routine pediatric physical examination should include careful examination of the breast and all palpable masses should be excised. Carcinoma occurring in the prepubertal child carries an improved prognosis particularly if the tumor stains PAS-positive. Carcinoma occurring in the postpubertal teenager probably carries the same prognosis as that in the adult and should be treated in a similar manner. The authors discuss such a case occurring in a 13-year-old girl.


Diagnostic accuracy of endoscopic retrograde cholangiopancreatography in hepatic, biliary, and pancreatic malignancy.

This paper reports the radiographic findings in 84 cases of biopsy-proven pancreatic and biliary malignancies studied by endoscopic retrograde cholangiopancreatography. In the 73 successful studies a diagnosis of tumor could be made in 67 patients (92%). The study was successful in 40 of 43 patients with pancreatic carcinoma and the diagnosis could be made in 37 on the basis of stenosis or obstruction of the ducts. There were 33 successful studies in the 41 patients with miscellaneous tumors including primary bile duct, ampullary, gallbladder, metastatic carcinoma, lymphoma, and hepatoma. A diagnosis of tumor was made in 30 studies. As has been observed in carcinoma of other hollow structures, the hallmark of malignancy in the pancreatic and biliary tract is obstruction and stenosis. This study indicates that malignant disease of the pancreas and tract is accurately assessed by this endoscopic method.


Role of ionophores in energy coupling.

If, as we deem inevitable, the principles of energy coupling are universal, then the necessity for charge-separating devices will apply across the board to all bioenergetic systems. Since, apart from the elctron transfer chain, ionophores are the only charge-separating devices available in bioenergetic systems, model of energy coupling that does not feature this central role of ionophores can be taken seriously. The ionophore approach thus opens the royal highway to the ultimate solution of all bioenergetic problems.


[Adrenal cortex and avitaminosis A: II. Complementary in vivo and in vitro studies on inhibition of corticoidogenesis in moderate deficiency in the rat. III. In vivo criterion for activity: glycemic response to ACTH].

II. We set up new experiments, both in vivo and in vitro, in order to explain the impaired adrenal response to stress, of vitamin A middly deficient, male Wistar rats. Injections of progesterone, 6 mg s.c. or i.p. every other day, did not improve the deficiency curse in our rats. In agreement with these results, we have found that, in vitro, the delta5-3beta hydroxysteroide dehydrogenase is not altered in deficient glands; we confirmed it histochemically. On the other hand, the 11beta and 18 hydroxylase activities are lowered in vitro (and presumably in vivo). When conditions of hyperactivity are established, the conversion of deoxycorticosterone (DOC) to 18 OH-DOC or to corticosterone is reduced and DOC accumulates in the incubation medium. III. When middly deficient intact rats are injected with ACTH i.p. (2UI p. 100 g of body weight), only some of them are found to have normoglycemia or hyperglycemia, 45 or 60 minutes after injection, as the controls do. Many deficient rats become hypoglycemic as the adrenalectomised animals do. We think that the changes in the glycemia after ACTH injection could be used to discriminate between the more or less deficient rats.


Current models of steroid hormone action: a critique.

In this review, we have traced the path of estradiol from its entry into the cell to the time of its release from the receptor. We feel that all of the current models are limited in one respect or another. We have examined most critically those currently most in vogue. The entry of estradiol into the cell is widely assumed to be simply a matter of diffusion. We have highlighted data that suggest the existence of a protein-mediated transport, but feel that the available data are too limited to make a definite conclusion.


Carcinomatous encephalomyelitis with auditory and vestibular manifestations.

A clinical and pathological study of carcinomatous encephalomyelitis is presented. Attention is drawn to the various types of nonmetastatic paraneoplastic syndromes and their particular association with oat cell carcinoma of the lung. The feature of special interest in this study is the onset with otologic symptoms, sudden deafness in the left ear and vertigo, at a time when the neoplastic basis for the disease was not clinically evident. The most striking change in the left temporal bone is the almost total loss of cochlear neurons in Rosenthal's canal and degeneration of both divisions of the vestibular nerve. The organ of Corti and stria vascularis are normal throughout the cochlear duct. The vestibular sense organs are normal. The left cochlear nucleus is devoid of neurons, this neuronal loss is accompanied by a well developed astrocytic and microglial response similar to that in the medulla and spinal cord. This represents a carcinomatous sensory neuropathy involving the left VIII nerve with simultaneous involvement of the left cochlear nucleus. The pathogenesis of this condition still defies explanation, but there are some insights in the autoimmune sector.


Susceptibility of beta-hemolytic streptococci to 65 antibacterial agents.

Tests for susceptibility of 29 group A, 4 group C, and 2 group G strains of beta-hemolytic streptococci to 63 antibiotics and to trimethoprim and sulfamethoxazole, singly and combined in a ratio of 1:16, were carried out in vitro. All strains tested were moderately or highly susceptible to all the antibiotics used except those belonging to the aminoglycoside and polymyxin groups. A few were also resistant to the tetracyclines and to sulfamethoxazole alone. Comparisons with results obtained in previous years indicate that, except for the tetracyclines and sulfonamides, there has been no change in the susceptibility of beta-hemolytic streptococci to the most important and useful antibiotics, particularly penicillin.


Antiviral activity of arabinosyladenine and arabinosylhypoxanthine in herpes simplex virus-infected KB cells: selective inhibition of viral deoxyribonucleic acid synthesis in synchronized suspension cultures.

The drug 9-beta-d-arabinofuranosyladenine (ara-A) significantly suppressed the formation of herpes simplex virus type 1-induced syncytia in BHK-21/4 cells at concentrations as low as 0.1 mug/ml. Optimal activity was noted when the drug was added before initiation of viral deoxyribonucleic acid (DNA) synthesis (3.5 h postinfection). The deaminated derivative of ara-A, 9-beta-d-arabinofuranosylhypoxanthine (ara-H), was at least 10 times less effective in suppressing the development of herpes simplex virus-induced syncytia. The replication of herpes simplex virus was measured by assaying fluids and cells from infected drug-treated cultures by using a plaque production technique. Ara-A at drug levels of >10 < 32 mug/ml completely blocked the replication of infectious virus particles. Ara-H was less effective than ara-A in reducing the replication of virions. Rates of host and viral DNA synthesis were monitored by pulse labeling herpes simplex virus-infected synchronized KB cells with [(3)H]thymidine and subsequently separating viral from cellular DNA in CsCl density gradients. During synthetic (S) phase, ara-A or ara-H at concentrations ranging from 3.2 to 32 mug/ml selectively inhibited viral DNA synthesis. At 3.2 mug of ara-A per ml, viral DNA synthesis was reduced 74% although total cellular DNA synthesis was unaffected. Increasing concentrations of ara-A produced increasing temporal delays in the maximal rate of host DNA synthesis. This time shift was not observed in cells treated with ara-H.


Inhibition of herpesvirus deoxyribonucleic acid and protein synthesis by tilorone hydrochloride.

Tilorone hydrochloride at a concentration of 10 mug/ml very efficiently inhibited herpes simplex virus growth in BSC1 cells when the virus is infected at a low multiplicity of infection. The adsorption of the virus was not affected by the drug, and the penetration of the deoxyribonucleic acid of the input virus into the cytoplasm and nuclei proceeded normally when tilorone hydrochloride was present. However, newly synthesized viral deoxyribonucleic acid was not detectable under these conditions, there was a remarkable decrease in the rate of viral polypeptide synthesis, and virus particles were not formed. The inhibition of herpesvirus growth by tilorone hydrochloride was absolutely dependent on the presence of the drug in the cultures. Pretreatment of the cells with the drug did not result in resistance to herpesvirus infection after the removal of the drug.


Inhibition of Clostridium perfringens by an antibiotic substance produced by Bacillus licheniformis in the digestive tract of gnotobiotic mice: effect on other bacteria from the digestive tract.

A strain of Bacillus licheniformis, established in the digestive tract of gnotobiotic mice, inhibited the subsequent establishment of a Clostridium perfringens strain ingested by the animals. This inhibitory effect depended on the in vivo production by B. licheniformis of an antibiotic substance having a number of the characteristics of bacitracin. If C. perfringens was the first to become established in the digestive tract of the gnotobiotic mice, B. licheniformis also became established but did not produce any antibiotic. Mutants of C. perfringens resistant to the antibiotic substance were not observed when the antibiotic was produced in situ by B. licheniformis, but were rapidly selected when the Bacillus culture filtrate or bacitracin was administered per os. B. licheniformis was also capable of eliminating from the digestive tract 5 of the 13 additional bacterial strains tested.


Action of polymyxin B on bacterial membranes: phosphatidylglycerol- and cardiolipin-induced susceptibility to polymyxin B in Acholeplasma laidlawii B.

To identify the polymyxin receptor molecules in the membranes of living microorganisms, fusion of intact Acholeplasma laidlawii B with lipid vesicles was investigated according to the procedure of Grant and McConnell (1973). The naturally polymyxin-resistant A. laidlawii B was treated with phospholipid vesicles prepared from purified phospholipids of the polymyxin-susceptible Salmonella typhimurium G30. A. laidlawii B absorbed between 15 and 45% of its own lipid content of the added tritium-labeled phospholipids without loss of viability. Association with the acidic components phosphatidylglycerol and cardiolipin produced a 10- to 30-fold increase in polymyxin susceptibility, which was not obtained with egg-phosphatidylcholine and mixed phosphatidylcholine-phosphatidylethanolamine vesicles. The polymyxin-sensitized cells bound 12 times more radioactive antibiotic than resistant cells. The phosphatidylglycerol-induced susceptibility was abolished by serum fraction V (Cohn) proteins.


Considerations on structure-activity relationships with mengovirus of substituted 5-amino-4-cyanopyrazoles.

In a screening program using the agar diffusion-plaque inhibition test, 22 out of 27 substituted aminopyrazoles tested showed marked antiviral activity against mengovirus in FL cell cultures. The efficacy of the different derivatives was compared by determining their effect on the yield of infectious virus after a one-step growth cycle. The antiviral activity against mengovirus correlated with the chain length of the substituents and increased or decreased according to the position of the side chain.


Mode of action in vitro against mengovirus of substituted 5-amino-4-cyanopyrazoles.

Substituted amino pyrazoles were found to exhibit plaque reduction and inhibition of the cytopathic effect of mengovirus on FL cells. Their antiviral activity was not caused by a virucidal effect or by inhibition of viral adsoprtion or penetration but by suppression of the virus multiplication. During a one-step growth cycle maximum suppression of virus yield occurred after compound addition from 0 to 2 h after infection. Progressively less viral inhibition appeared when compound was applied during the later part of virus replication. The antiviral effect was reversible by removal of the compound, and no inhibitor-resistant period occurred.


Use of sodium polyanethol sulfonate to selectively inhibit aminoglycoside and polymyxin antibiotics in a rapid blood level antibiotic assay.

Sodium polyanethol sulfonate inhibits aminoglycoside and polymyxin classes of antibiotics in direct proportion to its concentration. Aminoglycoside and polymyxin class antibiotics are selectively inactivated; penicillin, including the semisynthetic penicillins, cephalothin, chloramphenicol, clindamycin, tetracycline, erythromycin, and vancomycin are not inhibited. By incorporating sodium polyanethol sulfonate directly into the test medium it is possible, in a 4-h antibiotic blood level assay, to selectively obviate the activity of the aminoglycosides and polymyxins to determine the concentration of other antibiotics present in the same serum sample.


Stimulation of murine interferon by a substituted pyrimidine.

2-Amino-5-bromo-6-methyl-4-pyrimidinol (U-25,166) induced high levels of circulating interferon in mice when administered either parenterally or orally. Peak titers of interferon were found in the serum between 6 and 12 h after inoculation of the drug. Lower but significant levels of interferon were found in rat serum after administration of U-25,166 by either the intraperitoneal or oral route, and good levels of circulating interferon were observed in cats after oral treatment. Repeated intraperitoneal doses (50 mg/kg) of U-25,166 protected mice against intranasal encephalomyocarditis virus challenge. The minimal effective acute oral dose for antiviral activity was approximately 250 mg/kg. This was also the minimal dose that produced detectable levels of interferon. Maximum tolerated doses in mice were four to six times the minimal effective doses. A single oral treatment was protective in mice against challenge virus inoculated 24 h later. The compound protected mice from challenge with high levels of encephalomyocarditis virus, up to 20,000 mean lethal doses. Antiviral activity in mice was retained when certain minor substitutions were made in the U-25,166 structure.


Effect of cytosine arabinoside and 5-iodo-2'-deoxyuridine on a cytomegalovirus infection in newborn mice.

Murine cytomegalovirus was inhibited by 0.6 to 1.2 mug of cytosine arabinoside per ml and by 0.3 to 0.6 mug of 5-iodo-2'-deoxyuridine in mouse embryo fibroblast cells. Human cytomegalovirus was inhibited by similar concentrations of the two drugs in WI-38 cells. Intraperitoneal inoculation of suckling mice with 10(4.5) plaque-forming units of murine cytomegalovirus provides a model for disseminated human cytomegalovirus infection in human newborn infants and is characterized by a widespread infection involving the liver, spleen, lung, kidney, and brain with a 70 to 90% mortality in 7 to 9 days. Treatment with 12.5 mg of cytosine arabinoside per kg or 25 mg of 5-iodo-2'-deoxyuridine per kg twice daily for 8 days had no effect on final mortality or the pathogenesis of the infection with the exception of reduced viral titers in the spleen of 5-iodo-2'-deoxyuridine-treated animals. These data indicate that neither cytosine arabinoside nor 5-iodo-2'-deoxyuridine are effective in the treatment of murine cytomegalovirus infections in suckling mice and suggest that they may be of limited value in the treatment of severe cytomegalovirus infections in human neonates.


In vitro and in vivo activities of minocycline and other antibiotics against Acinetobacter (Herellea-Mima).

Minocycline was the most active of six antibiotics tested against 65 clinical isolates of Acinetobacter calcoaceticus (syn.: Herellea, Mima) received from six medical centers. In the Bauer-Kirby disk susceptibility test, all isolates were rated susceptible to minocycline, gentamicin, and polymyxin; 25% were resistant to tetracycline. In agar dilution tests, minocycline was two to four times more potent than gentamicin or polymyxin and eight times more potent than tetracycline. Ampicillin and cephalexin were relatively ineffective. Against lethal infections produced by five strains of A. calcoaceticus in mice, minocycline was, in general, more active than gentamicin or polymyxin on a dosage basis and significantly more active on a blood-level basis. Minocycline was significantly more potent than tetracycline on both dosage and blood-level bases against tetracycline-sensitive and -resistant strains. In the last decade there has been an increase in the reported incidence of acinetobacters in a variety of infections. The cultures are susceptible to few antibiotics. Our data show that minocycline could offer an effective alternative to the more toxic drugs for the treatment of these infections. Susceptibility should be determined with minocycline disks.


In vitro susceptibility of varicella zoster virus to adenine arabinoside and hypoxanthine arabinoside.

The in vitro susceptibility of varicella zoster (VZ) isolates to adenine arabinoside (ara-A) and hypoxanthine arabinoside (ara-Hx) was determined in human skin fibroblasts. Using a system in which deamination was inhibited, and using a modified plaque reduction method, the antiviral activities of ara-A and ara-Hx were separated. The plaque inhibitory concentration of ara-A for all VZ isolates tested was 1 to 2 mug/ml and 80 to 100 mug/ml for ara-Hx. A comparison of antiviral activity based upon total plaque suppression gave a rate of 40 to 50:1, in favor of ara-A.


Microbial metabolism of aryl sulphonates a re-assessment of colorimetric methods for the determination of sulphite and their use in measuring desulphonation of aryl and alkylbenzene sulphonates.

The reaction of 2,4-dinitroanilinomaleimide with sulphite which has been claimed as the basis of a suitable colorimetric assay for the anion was carefully re-examined. The sulphite-imide addition product provides a suitable and specific qualitative test for sulphite after separation by paper chromatography but the method as previously used is probably measuring the hydrolysis of the imide to 2,4-dinitroanilinomaleamic acid and cannot be used for sulphite determination either colorimetrically or in kinetic assays. A new colorimetric method for the determination of sulphite based on its reaction with Ellman's reagent, 5,5'-dithiobis(2-nitrobenzoic acid) is described and compared for sensitivity with the p-rosaniline-HCHO method. Both methods were used to show the formation of sulphite as the initial product of arylsulphonate metabolism by bacteria. The failure to find sulphite in similar cultures of a third organism was attributed to the very high activities of sulphite oxidase found in extracts. The Ellman reagent was examined as the basis of an indicator medium for the detection of sulphite-excreting colonies.


Activation of human leucocyte procollagenase by rheumatoid synovial tissue culture medium.

Production of a factor activating leucocyte procollagenase by tissue culture of rheumatoid synovium was shown. The activator was isolated, partly purified, and shown to be thermolabile, nondialysable, and had no activity toward casein, haemoglobin, histones, and PZ-peptide. The activity of the activator was partly decreased by trypsin.


Benign liver cell adenoma associated with use of oral contraceptive agents.

A benign hepatic adenoma in a young woman taking oral contraceptives for 7 years is reported. The diagnosis must be suspected in any young woman taking oral contraceptive agents who develop signs and symptoms of acute cholecystitis with hepatomegaly or mass, or when signs and symptoms of non-traumatic intraabdominal hemorrhage are present. Rupture of the tumor is a life-threatening complication. Treatment should be either hepatic lobectomy or wide local resection.


Cyclic adenosinemonophosphate in cerebrospinal fluid. Effects of theophylline, L-dopa and a dopamine receptor stimulant in rats.

The effects of L-Dopa and the dopamine receptor stimulant ET-495 on cisternal cAMP levels were studied in rats. L-dopa (100-200 mg/kg) increased cisternal cAMP levels by 60 to 80% of controls. When peripheral Dopa-decarboxylase was inhibited, smaller doses of L-Dopa were effective. Fla-63, an inhibitor of dopamine-beta-hydroxylase lowered the increase induced by L-Dopa which was completely suppressed by propranolol, not by phentolamine, suggesting that the cAMP increase is mediated through a central beta-adrenoceptor stimulation. ET-495 failed to influence cAMP levels which argues against a dopamine-sensitive adenylate cyclase involved in the L-Dopa effect. Moreover, large increases of cisternal cAMP were observed after treatment with theophylline , not papaverine which suggests different effects of these "phosphodiesterase inhibitors" on the cyclic AMP systems in the central nervous system or on transport mechanisms.


Intrathoracic paraganglioma arising from aorticosympathetic paraganglion.

A slowly growing intrathoracic paraganglion arising from an aorticosympathetic ganglion was removed from a 22-year-old woman. It was adherent to the parietal pleura and azygos vein, but not to the spinal cord. She has been well for the two years since that time. This patient represents the sixth tumor reported in this location. Most of the reported tumors are benign with occasional local invasion as seen in paragangliomas in other locations (carotid and aortic bodies). Complete excision of the tumor is the treatment of choice, but partial excision may provide long-term survival. A new classification and terminology suggested by Glenner and Grimley is emphasized. It groups logically the tumors arising from the paraganglion cells on the basis of anatomic distribution, innervation, and microscopic structure.


Cystosarcoma phyllodes metastatic to a brenner tumor of the ovary.

A patient had cytosarcoma phyllodes that developed metastases to bone and to a Brenner tumor of the ovary. The original breast tumor was reported as benign, but the patient died of metastases four months following mastectomy. The rapid growth in the ovary may have been due to estrogenic stroma in the Brenner tumor. This is the first report, to our knowledge, of cystosarcoma phyllodes metastasizing to another tumor.


[Study of cortico-melanotropic activities in the anterior pituitary of an anencephalic and a normal newborn].

Immunofluorescence studies by-means of anti-beta (1-24) corticotropin, anti-alpha (17-39) ACTH; anti-beta-MSH were performed on the anterior pituitary of an at-term anencephalic newborn. They showed small and rare cortico-melanotropic cells, as compared to those seen in the anterior pituitary of newborns. The comparison of these data with experimental observations suggest a possible autonomy of corticotropin function, although under hypothalamus control.


Grafting the unhealed perineal wound after coloproctectomy for Crohn disease.

The unhealed perineal wound following proctectomy for Crohn disease is a most unfortunate and disabling complication; treatment has been generally unsuccessful and disappointing. Since 1967, we have managed these wounds by skin-grafting the raw surface of the saucerized wound, rather than by attempting to obliterate the cavity. The graft is applied to the defect two or three days after wide debridement and saucerization. No attempt is made to cover more than the readily accessible raw surface. The graft is first dressed on the third postoperative day, and daily thereafter. The patient receives 40 units of corticotropin (ACTH) daily during his hospital stay; the drug therapy is continued in smaller dosage for several months. Results to date have been satisfactory, with seven wounds totally dry, and 37 resurfaced sufficiently so as to make the amount of drainage negligible. Four patients had results classified as poor.


The use of chicken tracheal organ cultures for the isolation and assay of avian infectious bronchitis virus.

A study has been made of the use of chicken tracheal organ cultures for the isolation and assay of avian infectious bronchitis (AIB) virus from both naturally and experimentally infected chickens. Six strains of AIB virus were investigated, 3 of which had been isolated from natural outbreaks of disease. Two of the virus isolations from the outbreaks of AIB were made directly into tracheal organ cultures without passage in embryonated eggs. Organ cultures prepared from 20-day-old embryos were used since they were found to be somewhat more sensitive in virus assay than those derived from chickens of up to 31 days of age. Ciliostasis, which was used as the marker of infectivity, was complete by 3 days after inoculation with each strain of virus examined. Virus could be isolated from both respiratory and non-respiratory tissue in tracheal organ cultures and these cultures were found to be at least as sensitive as 9-day-old embryonated eggs in detecting AIB virus either in pathological material or in serial dilutions. When virus was assayed in both systems, the titres were very similar. It is considered, therefore, that chicken embryo tracheal organ cultures offer a reliable alternative system to embryonated eggs for studying AIB virus.


In-vitro cultivation of human cytomegalovirus in thyroid epithelial cells.

Several strains of human cytomegalovirus including recent isolates, were grown in epithelial cells derived from thyroid tissue. All the strains tested grew in these cultures without pre-treatment of the cells, and no difference in cytopathic effect was detected between strains of genital and non-genital origin. 4 strains of CMV were isolated directly from urine in thyroid cells; however the failure to isolate 6 further strains in these cultures from other specimens may indicate that a higher multiplicity of infection is required to infect epithelial than fibroblast cells.


Simian virus 40-chinese hamster kidney cell interaction. II. The semipermissivity of the cell system.

Throughout in vitro passages, Chinese hamster kidney (CHK) cells progressively lost susceptibility to SV 40 virus infection while remaining continuously susceptible to viral DNA infection. Upon infection with SV 40 virus or viral DNA, the CHK cell line supported viral DNA and virus replication at a low level. SV 40 transformed CHK cell lines spontaneously produced small amounts of viral DNA and virions. The percentage of virus-producing cells was low. Various clones derived from each of these lines behaved as the parental cell population, leading to the conclusion that each CHK cell, whether transformed or not with SV 40, is potentially permissive for this virus.


Electron microscope observations on tubular structures in cells infected with herpes simplex virus type 2.

The fine structure of filamentous structures and lattice-like structures found in herpes simplex virus type 2-infected cells was studied. By specimen tilting experiments, it was demonstrated that the lattice-like structures were the cross-sectional views of the tubular structures and the filamentous structures were the longitudinal sections. The observation in negatively stained preparations of isolated tubular structures revealed that the structure was constructed by regular array of ring shaped elements. A hypothetical model is presented for tubular structures constructed on the basis of the present findings. The tubular structures appeared as early as 6 hours after infection and reached a maximum at 12 hours.


Heterogeneity of infectious bronchitis virus grown in eggs.

Egg-grown infectious bronchitis virus, strain Beaudette, was concentrated and centrifuged on sucrose density gradients to separate the virus into five peaks with densities of 1.144, 1.160, 1.172, 1.191 and 1.218 g/cm3. All peaks retained infectivity, complement fixation activity and were labelled with 3H-uridine. Morphologically the densest peak consisted of very large virus particles and amorphous material, the other peaks consisted of mainly intact particles although small differences in size and pleomorphism were seen. Polyacrylamide gel electrophoresis of material from the density gradient peaks revealed four major polypeptides and at least 10 minor polypeptides. The proportions of the polypeptides were approximately similar for all peaks with the exception of the densest peak in which the major polypeptides were greatly reduced. The four major polypeptides had approximate molecular weights of 1. 52,000, 2. 45,000, 3. 34,000, 4. 32,000. The major polypeptides 1 and 4 were shown to be glycosylated as were two of the minor polypeptides.


Characterization of cell lines derived from hamster tumors induced with the BK virus.

Some of the properties of three continuous cell lines derived from BK virus-induced hamster tumors were examined. The cell lines had in vitro growth characteristics of transformed cells. Morphologically most of the cells were fibroblastic, but multinucleated giant cells were also common. Ultrastructurally all three cell lines displayed the usual features of cells grown in vitro. Marked variation in the nuclear size and shape as well as prominent nucleoli were characteristic to these cells. No viruses or virus-like particles were found. Virus isolation attempts by fusing the cells with Vero cells were negative, and no virion antigen was detected in these cells by immunofluorescence. T antigen similar to that of other papovaviruses was found in the cells. This antigen stained with sera from a number of hamsters carrying transplanted BK virus-induced tumors, and also with SV 40 T antisera. The antigen disappeared after 30 minutes at 56 degrees C. Cytogenetic analyses showed that the three cell lines were heteroploid with subtetraploid numbers of chromosomes. Chromosome abnormalities were also seen. All three cell lines induced sarcomatous tumors in adult hamsters after subcutaneous inoculation.


Ultrastructural characterization and hepatic pathogenesis of duck plague virus.

Six-week-old white Pekin ducks were inoculated intravenously with duck plague virus (DPV) isolated from wild waterfowl. The virus replicated in hepatic macrophages, hepatocytes, and bile duct epithelium. In ultrathin sections, herpes-like nucleocapsids and virions were found respectively in the nucleus and cytoplasm of infected cells. Typical herpesviral capsids and virions were seen in negatively-stained preparations of duck embryo fibroblasts. Antibodies against Holland-attenuated strain of DPV reacted with virions of this isolate.


[Electron microscopy studies of the early stages of mosquito cell infection in vitro by the chilo iridescent virus and the infectious bovine rhinotracheitis (author's transl)].

The early stage of the viral infection in vitro of mosquito cells were studied by electron microscopy using a microculture technique. We demonstrate in this paper that two viruses: iridescent (CIV) and herpetic (Infectious bovine rhinotracheitis) can enter by viropexis and be uncoated in several lines of mosquito cells.


Cell-mediated immunity in Aujeszky disease virus infected pigs. I. Lymphocyte stimulation.

The appearance of cell-mediated immunity was studied in Aujeszky diseased pigs with the aid of the in vitro stimulation of sensitized lymphocytes. The first cell-mediated immunity reaction of lymphocytes occurred 4 days after infection. From day 7 to 35, the latest day tested, the reactions were most marked with lymphocytes from lymph nodes and spleen, whereas blood and thymus lymphocytes reacted less frequently; bone marrow lymphocytes showed no response. Reinfection did not considerably enhance lymphocyte reactivity. Humoral immunity was demonstrated a few days later than cell-mediated immunity. Neutralizing antibodies were first detected at day 7, reaching optimal titers at day 14. Complement fixing antibodies were detected from day 14 onward. Reinfection caused a very weak booster effect only on neutralizing antibody production. The sensitivity of the neutralization test could be enhanced up to sixfold by the addition of fresh guinea pig complement. It is concluded that cell-mediated immunity influences the early stage of infection with Aujeszky disease virus when humoral immunity is not yet demonstrable or yet rather low. Lymph nodes and spleen are apparently of special importance for the appearance of ADV-reactive lymphocytes.


Enhanced SV 40-virus replication in chinese hamster kidney cells pretreated with 5-iodo-2'-deoxyuridine.

SV40 virion production was enhanced when semipermissive Chinese hamster kidney cells were treated with 5-iodo-2'-deoxyuridine (IUDR), prior to infection either with virus or viral-DNA. Pretreatment did not increase SV40 replication in fully permissive monkey kidney and nonpermissive Syrian hamster or mouse embryo cells.


[Tissue nature of the lining of the lymph node sinuses].

The lymphatic nodes of intact albino rats were investigated electron microscopically. It was shown that the lymphatic sinuses were restricted by a layer of flattened cells; the basal membrane was absent. Certain distinctions in the structure of the cell lining sinuses and the reticular cells comprising the reticular base of the lymphoid tissue of the lymphatic node were found. The structure of the "sinus network" strands is shown. The structure of the cells of the sinuses lining is shown to be identical to the structure of cells of the vascular endothelium. It suggests the endothelial nature of the lining of the lymphatic node sinuses.


[Lesions of the central nervous system in adenovirus infections].

The investigation of the central nervous system in generalized adenoviral infection in two children showed the presence in large nuclei of the nerve cells of hyperchromic inclusions containing the antigen of adenovirus. This testifies to the possibility of developing encephalites caused by respiratory viruses.


[A histochemical study of the effect of progesterone on the carbohydrate metabolism enzymes of different morphologic forms of endometrial cancer].

The histochemical method was used for determining glycogen, phosphorylase, glycogensynthetase, and glucose-6-phosphatase in order to evaluate the effect of progesterone on different morphological forms of cancer of the endometrium in man. On the basis of histochemical analysis of changes in the carbohydrate metabolism in 29 cases of cancer before and after treatment with progesterone a conclusion is drawn that sensitivity of tumours of the endometrium to hormonotherapy only partially depends upon the degree of their morphological differentiation.


[The condition of mitochondria in experimental viral carcinogenesis].

A parallel histoenzymatic and electron-microscopy study of the state of mitochondria in experimental viral cancerogenesis induced by adenovirus of monkeys SA7(C8) and virus of Rous sarcoma in soft tissues of hamster was carried out. It was shown that the earliest changes in the cell in the process of its tumorous transformation were those in mitochondria. The character of deposits of diformazin at an early stage of viral cancerogenesis corresponded to the trend to the linear arrangement of mitochondria and to initial phenomena of their swelling. Progressive swelling of mitochondria in the process of tumorous growth was established, which correlated with histochemically revealed changes in the relationships of a number of reduction-oxidation enzymes.


[Malignant glomus tumors of the stomach].

A total of 16 malignant glomic tumours of the stomach were studied. It is established that these tumours, as well as benign formations, have organoid structure, but differ from the latter by disorders of the tissue architectonics, polymorphism of cells of various degree, occasionally insignificantly manifested. The clinical course of the tumours not always corresponds to their histological structure--in mature microscopic structure they are sometimes characterised by local destructive growth and can give metastases. The authors propose to differentiate among malignant glomic tumours of the stomach the same variants of the morphological structure as in benign ones, i.e., myoid, angioleiomyomatous, angiopericytic, and paraganglioma-like. The authors assume that malignant glomic tumours of the stomach appear as a result of malignization of benign tumours.


[Morphofunctional state of pulmonary macrophages and their biological protection during phagocytosis of toxic elements].

Studies of a morphofunctional state of macrophages of the lungs of rats in phagocytosis of quartz (size of particles 1-2 mum) and against the background of prophylactic administration of polyvinylridine-N-oxyde (PVNO) were carried out using the method of quantitative cytochemistry (interference microscopy and cytometry) and electron-microscopy techniques (raster and transmissive). It was established that administration of PVNO led to intensification of the phagocytic activity of macrophages, increase in the content of RNA and dry weight (total proteins) therein, enlargement of the sizes of cells. The electron-microscopy investigation showed a good preservation of organelles of macrophages.


Neurologic manifestations of glomus tumors in the head and neck.

In 75 patients with glomus tumors in the head and neck region, 57 tumors arose from the jugular bulb region, 11 from the middle ear, and seven from the vagus nerve. Thirty-seven percent (28 patients) had cranial nerve paralysis, and 14.6% (11) had intracranial tumor extension. The jugular foramen syndrome was associated with a 50% (two of four patients) incidence, and hypoglossal nerve paralysis with a 75% (three of four) incidence of posterior fossa tumor invasion. Horner syndrome and labyrinthine destruction had a 50% (two of four) incidence of a middle cranial fossa tumor invasion. The incidence of central nervous system (CNS) invasion with cranial nerve paralysis (excluding the seventh nerve) was 52% (11 of 21). Otologic findings and seventh nerve paralysis did not correlate with CNS tumor extension.


Palato-ocular synchrony during eyelid closure.

The effects of eyelid closure on oculomotor function was examined in 11 patients with palatal myoclonus. In eight patients, eyelid closure induced gross rhythmic vertical or almost oscillatory movements of the globes, which were synchronous with the rhythmic beat of the palatal myoclonus. The rhythmic vertical ocular movements induced by eyelid closure replaced calorically induced or spontaneous horizontal nystagmus present when the lids were open. Vertical ocular motions persisted during some stages of slow-wave sleep and reappeared during each stage of rapid eye movement. The physiologic basis of the palato-ocular synchrony may be similar to the eyelid closure, vertical eye movements, and palatal myoclonus that occurs in monkeys on stimulation of the central tegmental fasciculus.


The oncogenic potential of some avian adenoviruses causing diseases in chickens.

The oncogenic potentials of three different strains of avian adenoviruses (the Tipton strain of the inclusion body hepatitis virus, the DPI-2, and the Indiana C viruses) were investigated in newborn hamsters. Animal inoculations were via two routes, subcutaneous and intracerebral. All three viruses proved nononcogenic for the hamsters observed over a period of 225 days. However, lesions of hepatitis similar to those of inclusion body hepatitis of chickens were seen in three hamsters inoculated with the DPI-2 virus.


A lymphoproliferative disease in Japanese Quail.

A naturally occurring lymphoproliferative disease was found in 3 flocks of Japanese quail in Mexico. The tumorlike lesions were detected mainly in livers and spleens. Histopathology revealed a pleomorphic lymphoproliferative infiltration with dark-staining large cells, lymphocytes of different sizes, and reticulumlike cells. Marek's disease (MD) herpesvirus was not isolated from 74 quail tested, nor were antibodies to MD detected in 84 sera. Antibodies to reticuloendotheliosis (3/24) and avain leukosis virus subgroup A (2/24) were present. The possible etiology of this disease is discussed.


Identification of seven isolants of infectious bronchitis virus by interference with the B-1 isolant of Newcastle disease virus.

Seven isolants of infectious bronchitis virus (IBV) interfered with the development of hemagglutinins (HA) by the B-1 isolant of Newcaslte disease virus (NDV) in eggs previously inoculated with these IBV isolants. The I50 [defined as the minimum amount of IBV needed to produce, upon inoculation in embryonating chicken eggs (ECE), HA interference in 50% of these ECE reioculated 10 hr later with 10(2,41 EID50 (HA) of the B-1 strain of NDV] was less than 1 EID50. The interference was considered specific for IBV because if could be eliminated by addition of homologous anti-IBV serum. Furthermore, three other common poultry viruses (laryngotracheitis, fowlpox, and CELO viruses) did not interfere with NDV. In view of its sensitivity, specificity, and simplicity of performance, the IBV-NDV interference test appears to have considerable potential as a diagnostic test for IBV under field conditions.


New strains of infectious bronchitis virus isolated in Maine.

Twelve IBV isolants from diseased chickens in Maine were studied by serum-neutralization tests in chicken tracheal culture. Six isolants were neutralized by Mass-type antisera. The serum-neutralizing characteristics of six other isolants from vaccine breaks differed from any of the standard IBV viruses and appear to comprise three new strains, Maine 209, 212,and 246. The data provide additional evidence that strains of IBV responsible for disease probably vary from region to region and that the strains present must be known for a regional vaccination program to be effective.


Immunity to infectious bronchitis virus from spray vaccination with derivatives of a Holland strain.

Study was made of the protection of chickens from spray vaccination with different modifications of a Holland (H) strain of infectious bronchitis virus (IBV). Noted in each trial were the virus-serum-neutralizing antibody titers induced and the virulence of the vaccine virus at different passage levels. Protection against respiratory signs was good from challenge with a homologous virus serotype as well as with 5 heterologous IBV isolates. This occurred from vaccinating day-old chicks possessing homologous IBV antibodies as well as vaccinating susceptible 4-week-old chickens. The tracheal virus shed-rate following challenge was considered low (20% or less) with 4 of the challenge viruses. The challenge virus shed-rate was somewhat higher from vaccination at one day old than from vaccination at 4 weeks; the virus-serum-neutralization titers were lower, or even insignificant, with early-age vaccination. The immunogenicity and virulence of the vaccine virus was apparently reduced in the higher embryo passages employed. None of the vaccine derivatives was considered virulent under these conditions.


Isolation and identification of a turkey respiratory adenovirus.

A virus with physical and biological characteristics of an adenovirus was isolated from turkey poults with respiratory disease. The virus was ether-resistant and incorporated [3H] thymidine. Electron microscopy revealed virions of icosahedral configuration, approximately 78 nm in diameter, within the nuclei of infected cells. The virus produced cytopathology in turkey kidney cells, but did not produce observable disease when inoculated into commercial turkey poults or specific-pathogen-free embryonated chicken eggs. Virus-neutralization tests indicated widespread exposure to the virus in North Carolina turkey populations.


The influence of thymectomy on Rous sarcoma regression.

The incidence of regression of wing-web tumors induced by Rous sarcoma virus was shown to be dependent on the quantity of thymus tissue remaining after neonatal thymectomy in chickens of inbred line 6. Frequency of metastasis was associated negatively with the amount of thymus tissue present. Tumor regression and metastasis restriction both appeared dependent on the quantity of thymic tissue present.


Expression of the human adenylate kinase isozymes, phosphopyruvate hydratase, 6-phosphogluconate dehydrogenase, and phosphoglucomutase-1 in man-rodent somatic cell hybrids.

The expression of the adenylate kinase isozymes and of phosphopyruvate hydratase was studied in man-mouse and man-hamster hybrid clones. Concordant segregation of the loci coding for AK-2 and PPH was observed in 54 of 55 primary hybrid clones, and these loci were demonstrated to be synthetic with the loci specifying PGM-1 and PGD. The pattern of expression of the four enzymes in discordant clone suggests the gene order 1pter-(PGD, PPH)-AK-2-PGM-1-centromere. In addition, AK-1 was found to be expressed independently of AK-2.


Resolution and functional characterization of two mitochondrial iron-sulphur centres of the 'high-potential iron-sulphur protein' type.

Two distinct iron-sulphur centres of the 'HiPIP' (high-potential iron-protein) type are distinguished in both pigeon heart and ox heart mitochondria. These two species, although both are paramagnetic in the oxidized state, exhibit signals which differ in their detailed line shape, field position, and temperature- and power-dependence. They also exhibit different thermodynamic and kinetic behaviour and are located on opposite sides of the mitochondrial coupling membrane. One of these centres corresponds to Centre S-3. The other 'HiPIP'-type centre is removed readily from the mitochondrial membrane and its physiological function is not known.


Effects of cholecalciferol on the translocation of calcium by non-everted chick ileum in vitro.

An apparatus is described that allows perfusion of a non-everted segment of intestine in vitro and the study of the accumulation of substances within the mucosal cells. The translocation of Ca(2+) by rachitic-chick ileum and the effect of pretreatment with cholecalciferol was investigated, with the following conclusions. (1) Entry of Ca(2+) across the microvilli into mucosal cells is by diffusion; it does not require metabolic energy or the presence of any other inorganic ions. (2) Pretreatment of the chick with cholecalciferol causes increased permeability of the microvillus to Ca(2+) in both directions (lumen to cell, cell to lumen). The increased transport brought about by cholecalciferol in vivo can be partially mimicked by sodium dodecyl sulphate added in vitro. (3) The sign and the magnitude of the electrical potential difference prevailing across the ileum does not influence Ca(2+) transport. (4) Exit of Ca(2+) from the mucosal cell is temperature-sensitive, requires metabolic energy and Na(+). (5) Pretreatment with cholecalciferol caused increased movement of Ca(2+) out of the cell across the basement membranes. This effect of cholecalciferol given in vivo could be markedly increased by the presence of dicyclohexylcarbodi-imide in the perfusion fluid. These observations suggested that cholecalciferol increased Ca(2+) entry (and exit) at the mucosal surface and also caused Ca(2+) to be more available to the pump at the serosal surface.


Phosphatidate phosphohydrolase and palmitoyl-coenzyme A hydrolase in cardiac subcellular fractions of hyperthyroid rabbits and cardiomyopathic hamsters.

Activities of phosphatidate phosphohydrolase and palmitoyl-CoA hydrolase were determined in cardiac subcellular fractions prepared from rabbits which has received tri-iodothyronine and from hamsters with hereditary cardiomyopathy (strain BIO 14.6). 1. Both mitochondrial and microsomal fractions of hyperthyroid rabbit hearts produced 4-5 times as much diacylglycerol 3-phosphate from glycerol 3-phosphate and palmitate as did those of euthyroid hearts. 2. Phosphatidate phosphohydrolase, measured with phosphatidate emulsion, was activated by 1mm-Mg(2+) in all but the mitochondrial fraction of euthyroid rabbit hearts. The activation was more pronounced in subcellular fractions isolated from hyperthyroid hearts, so that the measured activities were significantly increased above those of the controls. The highest activity was found in the microsomal and lysosomal fractions. 3. In the absence of Mg(2+) during incubation, the difference in phosphohydrolase activities between eu- and hyper-thyroid states was not significant. 4. The phosphohydrolase of subcellular fractions of control hamsters did not respond to addition of 0.5-8.0mm-Mg(2+). The enzyme from cardiomyopathic hearts was slightly inhibited by this bivalent cation and therefore significant increases in activity were observed only in the absence of Mg(2+) from the assay system. 5. The rate of reaction by soluble phosphatidate phosphohydrolase was similar regardless of the nature of the substrate. Both when microsomal-bound phosphatidate was used as the substrate and when phosphatidate suspension was used, the activity of soluble enzyme was lower than that of the microsomal and lysosomal enzymes measured with phosphatidate suspension; this was especially so when the assay was carried out in the absence of Mg(2+). Neither tri-iodothyronine nor cardiomyopathy influenced the soluble phosphohydrolase activity in the two species. 6. Neither tri-iodothyronine nor cardiomyopathy significantly changed palmitoyl-CoA hydrolase activities in subcellular fractions. 7. Microsomal diacylglycerol acyltransferase and myocardial triacylglycerol content were also unchanged in the hyperthyroid state.


Regulation of mitochondrial protein turnover by thyroid hormone(s).

1. The effect of thyroidectomy on turnover rates of liver, kidney and brain mitochondrial proteins was examined. 2. In the euthyroid state, liver and kidney mitochondria show a synchronous turnover with all protein components showing more or less identical half-lives compared with the whole mitochondria. The brain mitochondrial proteins show asynchronous turnover, the soluble proteins having shorter half-lives. 3. Mitochondrial DNA (m-DNA) of liver and kidney has half-lives comparable with that of whole mitochondria from these tissues. 4. Thyroidectomy results in increased half-lives of liver and kidney mitochondria, with no apparent change in the half-life of brain mitochondria. 5. A detailed investigation of the turnover rates of several protein components revealed a significant decrease in the turnover rates of mitochondrial insoluble proteins from the three tissues under study. 6. The turnover rates of m-DNA of liver and kidney show a parallel decrease. 7. Thus it is apparent that thyroid hormone(s) may have a regulatory role in maintaining the synchrony of turnover of liver and kidney mitochondria in the euthyroid state. Turnover of brain mitochondria may perhaps be regulated by some other factor(s) in addition to thyroid hormone(s). 8. It seems likely that during mitochondrial turnover m-DNA and insoluble proteins may constitute a major unit. 9. The mitochondrial protein contents of the three tissues are not affected by thyroidectomy. 10. No correlation was seen between the turnover rate of mitochondria and cathepsin activity in any of the tissues under study in normal or thyroidectomized animals. 11. On the other hand, mitochondrial proteinase activity shows good correlation with the turnover rates of mitochondria in normal animals, and a parallel decrease in activity comparable with the decreased rates of turnover is observed after thyroidectomy. 12. It is concluded that mitochondrial proteinase activity may play a significant role in their protein turnover.


Gluconeogenesis by isolated guinea-pig liver parenchymal cells.

1. Guinea-pig hepatocytes were prepared by collagenase digestion of the perfused liver. 2. The highest rates of gluconeogenesis were obtained from fructose, followed by pyruvate, xylitol and lactate, glycerol and propionate in that order. Maximum rates of gluconeogenesis were attained at 6-10mm substrate. 3. An initial 15-min lag period occurred during gluconeogenesis from lactate. This lag was abolished by preincubating the cells or by preincubation plus the addition of NH(4)Cl or lysine. 4. The lactate/pyruvate and 3-hydroxybutyrate/acetoacetate ratios were increased during the lag and adjusted to values favouring rapid gluconeogenesis from lactate after 15min. 5. The data suggest that the low glucose synthesis during the lag resulted from a limitation of the glutamate-aspartate shuttle and from the unusual redox state of the NAD(+) couple prevailing during this period. 6. At 0.1mm, amino-oxyacetate, a transaminase inhibitor, decreased gluconeogenesis from lactate by 80%, but had a negligible effect on glucose production from pyruvate. Gluconeogenesis from lactate was also inhibited (20%) by 10mm-dl-3-hydroxybutyrate.


The release of adenosine 3':5'-cyclic monophosphate from the isolated perfused rat heart.

Although basal release of cyclic AMP from isolated perfused rat hearts was not measurable, isoprenaline induced substantial release of the nucleotide, suggesting that in vivo the myocardium can contribute to plasma cyclic AMP. Anoxia also increased the amount of cyclic AMP released, but insulin and nicotinate alone or in combination had no effect.


Analysis of metabolic changes in revertants of antibiotic-resistant micobacteria.

In the present report evaluation of the possible metabolic changes in partially revertants of M. tuberculosis var. hominis, reisolated from strains at high levels of drug-resistance after treatment with mutagenic agents, was reported. Based on the data presented here, it must be concluded that no significant biological and biochemical differences in mycobacterial cells revertant from high mutants mono-resistant to four aminoglycosides (Sm, Kana, Vio, Neo) and p-aminonasalicylic acid (PAS) are to be found.


[Seroepidemiologic research on the antipoliomyelitic immunity in adult unvaccinated subjects].

The Authors carried out a serological research on the polimyelitis viruses in 727 adult subjects who had not been vaccinated orally. The results of the titration of the neutralizing antibodies showed that the situation of immunity with respect to poliomyelitis is still satisfactory. However, the difficulty of making an exact estimation of the duration of the state of immunity to poliomyelitis, and the persistent, though reduced, circulation of wild polioviruses are such that a continuous epidemiological control is advisable.


[ADAM syndrome].

We present six cases of ADAM syndrome, that we have been studying at the Unit of Clinical Genetics of the Hospital Infantil de México. We wish to stress how it is possible using medical photography to appreciate the different characteristics of ADAM syndrome: A-Amniotic, D-Deformation, A-Adhesions, M-Mutilation.


Treatment of transplanted rat tumours with double-stranded RNA (BRL 5907). I. Influenced of systemic and local administration.

Growth of transplanted rat tumours was retarded and in some cases completely suppressed when cells were injected subcutaneously in admixture with double stranded RNA (ds-RNA). This response required intimate contact between ds-RNA and tumour cells and systemic treatment with the agent failed to prevent progressive growth of a range of rat tumours. Direct cytotoxic effects of ds-RNA may contribute to tumour suppression since the compound was cytotoxic in vitro for cultured tumour cells. The involvement of host factors is suggested, however, by the in vivo tests showing variations in susceptibility to ds-RNA mediated tumour suppression similar to that previously observed with bacterial adjuvants.


Treatment of transplanted rat tumours with double-stranded RNA(BRL 5907). II. Treatment of pleural and peritoneal growths.

Intrapleural growth of transplanted rat tumours was prevented or retarded by intrapleural administration of double-stranded RNA. A similar suppression of growth was achieved with peitoneal tumours by the intraperitoneal injection of the compound. These studies indicate the possible potential of this form of treatment of thoracic and peritoneal tumours for clinical application in the treatment of mesothelioma.


Studies on the relationship between concanavalin A and SV40-transformed human fibroblasts.

The extent of binding of 125I-Con A to the surface of SV40-transformed human fibroblasts and the degree of agglutination of the cells by the native lectin have been measured. In addition, trypsinized and succinylated Con A have been used to study the effects of the lectin upon certain cell growth parameters. Trypsinized Con A was found to alter the growth rate, the saturation density and the contact inhibition of the transformed cells, an effect not neutralized by alph-methyl-D-mannoside.


Recovery of herpes simiae (B virus) from both primary and latent infections in rhesus monkeys.

The suspected ability of herpes simiae (B virus) to persist in a latent form has been confirmed in rhesus monkeys. The virus was recovered from primary oral lesions of 2 young monkeys and again, 6 months after disappearance of symptoms, from cultures of Gasserian ganglia taken from the same individuals. B virus was identified by its effects in vivo and in vitro and in cross neutralization tests with antisera to reference B virus and herpes simplex virus. Tests showed that the same virus was present in oral lesions and in ganglia. The one-way immunological relationship between herpes simplex virus and B virus was clearly shown in results of cross neutralization tests.


Latency of Herpes Simiae (B virus) in rabbits.

When small doses of herpes simplex virus (HSV) were given to rabbits a significant degree of protection was afforded against later infection with herpes simiae (B virus). Only 12/47 rabbits died within 3 weeks of receiving B virus dosages that would normally have proved lethal. B virus became latent in survivors and was recovered from brain and spinal cord suspensions from rabbits which died within 6 months and from dorsal root ganglia of rabbits which had survived for more than 2 years without overt signs of infection. A minority of the survivors tested also yielded HSV. Rabbits with latent B virus in the ganglia showed little or no detectable neutralizing antibody to B virus. The possibility is discussed that human populations having a high frequency occurrence of HSV antibody may include carriers of latent B virus.


Ultrastructural changes in livers of tumour bearing rats.

An ultrastructural study of the livers of rats bearing transplanted mammary carcinomata from 4 to 12 weeks after transplantation revealed changes mainly in cytoplasmic structure of hepatocytes. The significant findings were pleomorphism of mitochondria, presence of large number of myelin figures, marked increase in microbodies, dilatation of rough endoplasmic reticulum, proliferation of smooth endoplasmic reticulum, intracytoplasmic cholesterol clefts and absence of increase in lysosomes. These alterations have been compared with that of others who have used different animal model systems to study the effect of tumour in livers of host. Centrioles in Küpffer cells are described for the first time in this study.


Effects of 8-substituted adenosine 3',5'-monophosphate derivatives on high Km phosphodiesterase activity.

Most of twenty-one 8-substitued adenosine 3',5'-monophosphate derivatives were found to inhibit competitively the hydrolysis of adenosine 3'5'-monophosphate by partially purified high Km (Michaelis-Menten constant) phosphodiesterase from hog brain cortex, which had one active site at high concentration of adenosine 3',5'-monophosphate (0.3 to 4.0 mM). The Ki value for the 8-substituted alkylaminoadenosine 3'5'-monophosphate derivative was found to decrease with increasing unbranched carbon chain of the substituent, and a minimum value was obtained in the case of 8-octylaminoadenosine 3',5'-monophosphate. The Ki value, however, increased gradually as the substituent of derivative became longer than that of 8-octylminoadenosine 3'5'-monophosphate. The similar phenomenon was observed in the 8-substituted alkylthioadenosine 3',5'-monophosphate. The standard affinity for adenosine 3,5'-monophosphate of the high Km phosphodiesterase was 5.0 kcal/mol, which was calculated from Km. The standard affinity for 8-hexylthioadenosine 3',5'-monophosphate, which inhibited most strongly the enzyme activity, was 7.2 kcal/mol. The difference (2.2 kcal/736) between the standard affinity for adenosine 3',5'-monphosphate and that for 8-hexylthioadenosine 3',5'-monophosphate seems to be based on the partial affinity for the substituent (hexylthio group) of the active site on the enzyme or its neighborhood. A characteristic similar interrelation between substituent length of derivatives and their inhibitory effect on the enzyme activity was observed similarly in two different series of derivatives, 8-substituted alkylaminoadenosine 3',5'-monophosphate and alkylthioadenosine 3',5'-monophosphate. The results may indicate the characteristic structure of the active site of the enzyme or its neighborhood.


Molecular forms of acetylcholinesterase from Torpedo californica: their relationship to synaptic membranes.

The 16S and 8S forms of acetylcholinesterase (AchE), which are composed of an elongated tail structure in addition to the more globular catalytic subunits, were extracted and purified from membranes from Torpedo californica electric organs. Their subunit compositions and quaternary structures were compared with 11S lytic enzyme which is derived from collagenase or trypsin treatment of the membranes and devoid of the tail unit. Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the absence of reducing agent, appreciable populations of monomeric through tetrameric species are observed for the 11S form. Under the same conditions, the 16S form yields only monomer and dimer in addition to a higher molecular weight species. If complete reduction is effected, only the 80,000 molecular weight monomer is dominant for both the 11S and 16S forms. Cross-linking of the 11S form by dimethyl suberimidate followed by reduction yields monomer through tetramer in descending frequency, while the 16S form  again shows a high molecular weight species. A comparison of the composition of the 11S and 16S forms reveals that the latter has an increased glycine content, and 1.1 and 0.3 mol % hydroxyproline and hydroxylysine, respectively. Collagenases that have been purified to homogencity and are devoid of amidase and caseinolytic activity, but active against native collagen, will convert 16S acetylcholinesterase to the 11S form. Thus, composition and substrate behavior of the 16S enzyme are indicative of the tail unit containing a collagen-like sequence. A membrane fraction enriched in acetylcholinesterase and components of basement membrane can be separated from the major portion of the membrane protein. The 16S but not the 11S form reassociates selectively with this membrane fraction. These findings reveal distinct similarities between the tail unit of acetylcholinesterase and basement membrane components and suggest a primary association of AchE with the basement membrane.


The monosaccharide composition and sequence of the carbohydrate moiety of human serum low density lipoproteins.

Human serum low density lipoprotein (d = 1.027-1.045) was delipidated with organic solvents and the apoprotein digested with thermolysin. The digest was fractionated by gel filtration and DEAE-cellulose chromatography. Two glycopeptides were obtained. One of the glycopeptides (GP-I) contained 2 residues of N-acetylglucosamine and 6 residues of mannose per mole of the glycopeptide, while the other contained 2 sialic acid, 5 mannose, 2 galactose, and 3 N-acetylglucosamine residues per mole of glycopeptide. The results of sequential enzymatic digestion with purified glycosidases, periodate oxidation, and partial acid hydrolysis lead us to propose the following sturctures for the two glycopeptides: (see article). These glycopeptides represent at least 50% of the carbohydrate moiety of LDL.


Inactivation and reactivation of mitochondrial respiration by charged detergents.

Respiration of submitochondrial preparations can be inhibited by the cationic detergent cetyl trimethyl ammonium bromide and the anionic detergent sodium dodecyl sulfate in the range of 0.3-2 mumol of detergent per mg of mitochondrial membrane protein depending on the substrate and detergent used. This inhibition can be rapidly reversed by neutralizing a given detergent by the detergent of the opposite charge. At higher levels of the inhibiting detergent, no such reactivation was observed. Spin labeling assays of membrane structure were used to correlate structural effects with the loss and recovery of respiratory functions. Because the detergents progressively disrupt membrane structure, mitochondrial were cross-linked with bifunctional imidoesters to an extent that osmotic properties and detergent lysis were gone, but respiration remained. Such fixed respiring mitochondria also show inhibition reactivation phenomena.


Energy transduction in photosynthetic bacteria. X. Composition and function of the branched oxidase system in wild type and respiration deficient mutants of Rhodopseudomonas capsulata.

The respiratory chain of Rhodopseudomonas capsulata, strain St. Louis and of two respiration deficient mutants (M6 and M7) has been investigated by examining the redox and spectral characteristics of the cytochromes and their response to substrates and to specific respiratory inhibitors. Since the specific lesions of M6 and M7 have been localized on two different branches of the multiple oxidase system of the wild type strain, the capability for aerobic growth of these mutants can be considered as a proof of the physiological significance of both branched systems "in vivo". Using M6 and M7 mutants the response of the branched chain to respiratory inhibitors could be established. Cytochrome oxidase activity, a specific function of an high potential cytochrome b (E'0 = +413 mV) is sensitive to low concentrations of KCN (5-10(-5) M); CO is a specific inhibitor of an alternative oxidase, which is also inhibited by high concentrations of KCN (10(-3) M). Antimycin A inhibits preferentially the branch of the chain affected by low concentrations of cyanide. Redox titrations and spectral data indicate the presence in the membrane of three cytochromes of b type (E'0 = +413, +260, +47 vM) and two cytochromes of c type (E'0 = +342, +94 mV). A clear indication of the involvement in respiration of cytochrome b413, cytochrome c342 and cytochrome b47 has been obtained. Only 50% of the dithionite reducible cytochrome b can be reduced by respiratory substrates also in the presence of high concentrations of KCN or in anaerobiosis. The presence and function of quinones in the respiratory electron transport system has been clearly demonstrated. Quinones, which are reducible by NADH and succinate to about the same extent can be reoxidized through both branches of the respiratory chain, as shown by the response of their redox state to KCN. The possible site of the branching of the electron transport chain has been investigated comparing the per cent level of reduction of quinones and of cytochromes b and c as a function of KCN concentrations in membranes from wild type and M6 mutants cells. The site of the branching has been localized at the level of quinones-cytochrome b47. A tentative scheme of the respiratory chains operating in Rhodopseudomonas capsulata, St. Louis and in the two respiration deficient mutants, M6 and M7 is presented.


The branched respiratory system of photosynthetically grown Rhodopseudomonas capsulata.

Various respiratory electron transport activities of Rhodopseudomonas capsulata were studied in membrane fragments prepared from photosynthetically grown cells of a parental strain and two terminal oxidase-defective mutant strains. The NADH and succinate oxidase activities of the mutant having a functional N,N,N1,N1-tetramethyl-p-phenylenediamine oxidase, M6, were consideraly more sensitive to inhibition by either antimycin A or cyanide than the corresponding activities of the mutant lacking a functional N,N,N1,N1-tetramethyl-p-phenylenediamine oxidase, M7. The parental strain, Z-1, but not the mutants, showed biphasic inhibitory responses of NADH and succinate oxidase activities with either antimycin A or cyanide. In certain reactions no differences in inhibitor susceptibility were found among the strains tested, implying that the pathways involved were unaffected in the mutants. In this category were the actions of rotenone on NADH oxidase, antimycin A on cytochrome c reductase and, in M6 and Z-1, cyanide on N,N,N'N'-tetramethyl-p-phenylenediamine oxidase. These results suggest that the respiratory chain of the parental strain branches at the ubiquinone-cytochrome b region into two pathways, each branch goes to a distinct terminal oxidase, and either may be blocked independently by genetic mutation.


A flash photolysis ESR study of photosystem II signal IIvf, the physiological donor to P-680+.

In flash-illuminated, oxygen-evolving spinach chloroplasts and green algae, a free radical transient has been observed with spectral parameters similar to those of Signal II (g approximately 2.0045, deltaHpp approximately 19G). However, in contrast with ESR Signal II, the transient radical does not readily saturate even at microwave power levels of 200 mW. This species is formed most efficiently with "red" illumination (lambda less than 680 nm) and occurs stoichiometrically in  a 1:1 ratio with P-700+. The Photosystem II transient is formed in less than 100 mus and decays via first-order kinetics with a halftime of 400-900 mus. Additionally, the t1/2 for radical decay is temperature independent between 20 and 4 degrees C; however, below 4 degrees C the transient signal exhibits Arrhenius behavior with an activation energy of approx. 10 kcal-mol-1. Inhibition of electron transport through Photosystem II by o-phenanthroline, 3-(3,4-dichlorophenyl)-1,1-dimethylurea or reduced 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone suppresses the formation of the light-induced transient. At low concentrations (0.2 mM), 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone partially inhibits the free radical formation, however, the decay kinetics are unaltered. High concentrations of 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (1-5 mM) restore both the transient signal and electron flow through Photosystem II. These findings suggest that this "quinoidal" type ESR transient functions as the physiological donor to the oxidized reaction center chlorophyll, P-680+.


Zero field resonance and spin alignment of the triplet state of chloroplasts at 2 degrees K.

Microwave induced transitions in zero magnetic field have been observed in the photoinduced triplet of chloroplasts treated with dithionite by monitoring changes in the intensity of the 735 nm fluorescence band at 2 degrees K. Similar results were obtained with chloroplasts treated with hydroxylamine plus 3-(3,4-dichlorophenyl)-1,1-dimethylurea and preillumination. The zero field parameters are D = 0.02794 +/- 0.00007 cm-1, E = 0.00382 +/- 0.00007 cm-1, i.e. equal to those of monomeric chlorophyll a to within the experimental error. The photoinduced triplet appears to be linked to Photosystem II. This indicates that the low temperatures 735 nm fluorescence band of chloroplasts is at least partly due to Photosystem II.


Uptake and degradation of low density lipoprotein by swine arterial smoot muscle cells with inhibition of cholesterol biosynthesis.

We have previously proposed on the basis of studies in hepatectomized animals that low density lipoproteins are degraded at a significant rate by peripheral tissues. To test the capacity of one peripheral cell type to catabolize low density lipoprotein, cultures of swine aortic smooth muscle cells were incubated with homologous 125I-labeled low density lipoprotein and uptake and degradation measured. Degradation of 125I-labeled low density lipoprotein to products soluble in trichloroacetic acid showed an initial lag period of 1--2 h after which the rate increased and remained linear for the following 15 h. Rates of degradation increased sharply with low density lipoprotein concentration over the lower range (from 0--25 mug protein/ml) and then more slowly up to the highest concentration tested, 300 mug protein/ml. Even at very low concentrations, 1 mug low density lipoprotein protein/ml (less than 10% of the plasma low density lipoprotein concentration), the in vitro degradation rate (per kg of smooth muscle cells) exceeded the in vivo degradation rate (per kg of total body weight). To the extent that smooth muscle cells are representative of other peripheral cells, the results support the proposal that peripheral degradation of low density lipoprotein apoprotein may be quantitatively important. The rate of incorporation of labeled acetate into sterols was suppressed in cells incubated with whole serum, low density and very low density lipoproteins, or suspensions of free cholesterol. In this respect, the results were similar to those observed in human skin fibroblasts studied concurrently. However, high density lipoprotein inhibited sterol synthesis by about 25% in swine smooth muscle cells while it had no effect in human skin fibroblasts.


The effect of hypoxia on uptake and degradation of low density lipoproteins by cultured human arterial smooth muscle cells.

Atheroma have been produced in experimental animal by systemic hypoxia. This study assessed the effects of hypoxia on binding, uptake and degradation of human low density lipoprotein (LDL) by human arterial smooth muscle cells, the cell involved in atherogenesis. The LDL content of the smooth muscle cell grown in the usual conditions (95% air [20% O2], 5% CO2) increased with the incubation time of LDL in the medium (7.5 mug protein/ml of medium); the trypsin releasable LDL "binding" reached a plateau by 24 h (2.2 +/- 1.3 [x +/- S.D.]) ng/mug LDL protein added per 10(6) cells whereas the LDL in the cell after trypsinization ("net uptake") continued to increase up to 48 h (6.5 +/- 4.6 ng/mug LDL protein added per 10(6) cells at 48 h). LDL protein degradation increases rapidly between 7 and 48 h (10.4 ng/mug LDL protein added per 10(6) cells at 24 h) after an initial delay of approximately 7 h. Smooth muscle cells grown under hypoxic conditions (5%02) had similar LDL "binding " but showed increased "net uptake" (10.7 +/- 4.8 ng/mug LDL protein added per 10(6) cells) and a 36 +/- 13% decrease in degradation (p less than 0.05; n =8). The impaired degradation of lipoprotein by smooth muscle cells may, in part, explain the role of hypoxia in atherogenesis.


The content of prostaglandin E and prostaglandin F2alpha in the exudate of carrageenin granuloma of rats.

1. Granuloma was made by the subcutaneous injection of 2% carrageenin solution on the dorsum of male rats. Eight, 16, 24 and 72 h after the injection. the exudate from each rat granuloma was withdrawn and extracted for rpstaglandins. 2. Extracted prostaglandins were separated prostaglandin E and prostaglandin F group by silicic acid mini-column chromatography. Then the amount of prostaglandin E and prostaglandin F2alpha were determined by the radioimmunoassay method. 3. The levels of prostaglandin E in the granuloma exudates were 4.6 ng/ml at 8 h after the carrageenin injection, then decreased 3.6 ng/ml and to 1.1 ng/ml at 16 h and 24 h, respectively. Seventy-two h after the injection, prostaglandin E level was increased to 8.1 ng/ml. 4. The levels of prostaglandin F2alpha in the exudate were as follows: At 8 h after the carrageenin injection, the level was 9.4 ng/ml, then decreased to 1.3 ng/ml and to 0.8 ng/ml at 16 h and 24 h, respectively. Seventy-two h after the carrageenin injection, it was again elevated to 4.7 ng/ml. 5. The exudate of granuloma, 24 and 72 h after the carrageenin injection, was incubated with [3H]prostaglandin E1 at 37 degrees C for 30 min. Then the acidic ether extract was subjected to reversed phase partition chromatography. It was found that the exudate of 24 h and 72 h granuloma had little activity of prostaglandin 15alpha-hydroxy dehydrogenase.


Combined enzymatic and chemical approaches to the synthesis of unique polyribonucleotides.

The enzymatic polymerization by polynucleotide phosphorylase of 6-chloro-9-(beta-D-ribofuranosyl)purine 5'-diphosphate to poly(6-chloropurinylic acid) and its conversion to poly(6-thioninosinic acid) is described. The sulfur isostere of poly(I) was found not to form a complex with poly(C), but to form a self-association complex with a Tm around 295 degrees K. The sedimentation velocities,  pKa and Tm values of the polymer have been examined under various conditions. A two (or more) stranded helical array is suggested as the most probable structure. Thermal loss of the thione chromophore was noted for poly- (S6I), S6IMP and S6I; the degradation product from S6I was shown to be inosine.


Stimulatory actions of thyrotropin and dibutyryl cyclic AMP on transcription and translation in the regulation of thyroidal protein synthesis.

When beef thyroid cells were incubated with thyrotropin and then tested for protein synthesizing activity, the stimulatory effect of thyrotropin appeared in two distinct phases: first, an immediate stimulation which continued as long as thyrotropin was present, but died away promptly after withdrawal of the added thyrotropin; and second, a delayed stimulation with a lag period of 1 to 2 h, which persisted after thyrotropin withdrawal. The fast and the delayed effects each stimulated protein synthesis by about 25%, so that after 4 h of thyrotropin treatment, total stimulation amounted to 50% above basal levels of activity. The fast thyrotropin effect was not affected by actinomycin D or cordycepin, and hence is evidently not dependent upon induced RNA synthesis. In contrast, the delayed thyrotropin effect was completely prevented by actinomycin D or cordycepin. These findings suggest that the fast effect of thyrotropin occurs at the translational level to provide for acute adjustments in thyroglobulin production by regulating the rate of translation of existing mRNA. The delayed thyrotropin effect, by initiating certain transcriptional reactions could be the means for selectively inducing the production of specific enzymes or proteins. Both the fast and delayed effects of thyrotropin were faithfully reproduced by dibutyryl cyclic AMP. This finding seems to be the first in vitro demonstration of a cyclic AMP induced transcriptional response to a hormone in mammalian cells.