Catheter technique for encephalography.

Catheter technique for encephalography. A catheter technique for encephalography utilizing a polyethylene catheter is described. The advantages are principally that it reduces the possibility of needle displacement during examination and thus the danger of extra-arachnoid injection of air.


Hypocycloid tomography in encephalography to demonstrate deep hemorrhage of the brain. Five cases of spontaneous deep brain hemorrhage have been presented. Two hemorrhages occurred above the tentorium and three below. The advantages of encephalography combined with tomography in diagnosing and following patients with deep brain hemorrhage are demonstrated in each case.


Secondary empty sella syndrome.

Secondary empty sella syndrome. Etiology and pathogenesis of empty cella syndrome remains undetermined in some cases (primary or idiopathic type), while in others it is related to treatment of pituitary and parasellar disease (secondary type). Ten patients with empty sellae, secondary to treatment of pituitary adenomas (five cases), granulomas (three cases), craniopharyngioma (one case), and optic glioma (one case) are presented. Pathogenesis of empty sella is discussed and the importance of neuroradiologic evaluation of this syndrome is stressed.


[Prolactin-secreting adenomas. Radiologic signs]. The sellar and tomo-encephalographic changes in pituitary prolactin secreting adenomas are described on the base of 75 cases detected within a period of 2 years. In the diagnosis of smaller adenomas in which the sellar findings may be questionable, the value of fractionated encephalography is emphasized, and especially of frontal tomograms, in order to demonstrate a lateral bulging of the sellar diaphragm and a shift of the pituitary stalk to the opposite side.


Encephalography using supra-occipital tapping of the cisterna magna. A report is given of the results of supra-occipital tapping of the cisterna magna and dynamic encephalography by that route. In 72 patients with cranial hypertension this encephalography was well tolerated. Supra-occipital tapping is less dangerous than suboccipital puncture as regards the possibility of injury to the medulla. In 38 cases, the method led immediately to the diagnosis of herniation of the tonsils.


[Effect of fractionated pneumoencephalography on intracranial pressure]. The major changes in clinical condition during and after encephalography may be explained by intracranial pressure variations. If neither intracranial hypertension nor CSF dynamic perturbations exist, minimal complications may be predicted, but in intracranial hypertension immediate deterioration may occur. Despite an apparently good level of tolerance, complications may develop if the compensation phenomena cannot occur: normal pressure hydrocephalus, craniosynostosis. In the cases with good clinical tolerance, minimal changes occur when air injection is administered at a slow rate and with a small volume.


Spontaneous displacement of air after encephalography. In some cases it has been observed that after encephalography air seems to be displaced from one part of the ventricular system to another without movements of the head. This apparently occurs in spite of the law of upward displacement of gases in fluids.


Ventriculography with water-soluble contrast media. Ventriculography with water-soluble contrast media is described. The advantage of subtraction and tomography is discussed. The incidence of side effects and complications of ventriculography with air and Dimer-X are given and compared with data about Conray 60.


The lateral recess of the fourth ventricle examined with a water-soluble contrast medium. Difficulties still remain in the routine radiologic examination of the lateral recesses of the fourth ventricle. Ventriculography with the direct introduction of water-soluble contrast medium into the third ventricle has improved the results. The normal anatomic and radiologic appearances and pathologic changes of the lateral recesses are discussed.


Central ventriculography in childhood with water-soluble contrast media. Central ventriculography was performed by catheterization of the third ventricle or puncture of the anterior horn of the lateral ventricle through the anterior fontanelle. Three different contrast media were compared in 138 cases. Densopax offered the following advantages over Conray and Triyosom 60 MG: Less discomfort to the patient, the procedure may be performed with the patient supine, no danger of embolization, and better demonstration of the aqueduct and fourth ventricle when the ventricular system is greatly dilated.


[Intraspinal venous hypertension due to multiple anomalies in the caval system. A major cause of myelopathies]. Increased venous intraspinal pressure is described as a venous system disease, resulting in numerous unexplained paraplegias and tetraplegias. The chronic venous stasis in the intraspinal plexuses, into which the circulation of the spinal cord is drained, is due to the association of multiple abnormalities (stenoses, compressions, thromboses) on the major pathways of the caval and azygos system. The abnormalities, most of which are not known, are demonstrated by a special procedure, the cavo-spinal phlebography, and some of them are subjected to surgery.


[Cavo-spinal phlebography in myelopathies of venous origin. Application of the method in 115 cases]. The intraspinal venous stasis, described by ABOULKER as the cause of numerous myelopathies, is due to the addition of multiple venous abnormalities, demonstrated by cavospinal phlebography. The venae cavae and their major affluents and the prespinal system (lumbar and ascending lumbar veins, azygos, hemi-azygos, right superior intercostal and vertebral veins) are explored by catheterization. Cavo-spinal phlebography reveals multiple obstacles and the resulting stasis in the intraspinal plexus.


[Cavo-spinal phlebography in myelopathies. Stenoses of internal jugular and azygos veins, venous compressions and thromboses]. Increased intraspinal venous pressure, resulting according to ABOULKER in numerous spastic paraplegias and quadriplegias is due to multiple venous abnormalities demonstrated by cavo-spinal phlebography. The most frequent are stenoses of the internal jugular veins, the left renal, the left iliac veins, the azygos veins and compressions of the innominate venous trunks. These abnormalities cause a permanent stasis in the intraspinal plexuses through excessive supply or insufficient drainage. Out of 80 patients, 60 per cent had at least 2 abnormalities, 38 per cent at least 3 abnormalities.


Lumbar tomomyelography with water-soluble contrast medium. Myelography supplemented by tomography was routinely carried out in 150 cases. The number of false negative findings was reduced practically to nil. At present, this method is only used in cases with equivocal results at conventional myelography or with poor correlation between clinical symptomatology and myelography. The frequency of side effects in 500 examinations with Dimer-X is unimportant.


Xeroradiography. Its value in gas myelography. Electrostatic imaging with its capacity for enhancing borders improves radiographic demonstration of gas-soft tissue interfaces in gas myelography. Its wide recording latitude also provides satisfactory delineation through areas of widely varying photon absorption. Xeroradiogrphy surmounts the paradox wherein contrast and latitude are inversely related so that the combination of both high contrast and broad latitude are simultaneously used to advantage. The xerogram, especially in the thoracic region, may be sufficiently diagnostic so that tomography is not necessary. Where additional information is required, tomographic xeroradiography can provide diagnostic information not perceptible on film tomograms.


Orbital phlebography in evaluation of the cavernous sinus and adjacent basal veins of the skull. Frontal vein phlebography has proven to be of great diagnostic value in the evaluation of pituitary tumours and of lesions in the vicinity of the sella turcica. It permits the diagnosis of venous thrombosis; similar phlebographic changes may be the only positive radiologic finding in cases with malignant tumour of the skull base. Phlebography is thus to be recommended as an early method to be used in possibly malignant lesions at the skull base.


Postural scintimyelography and dynamic CSF-pressure measurements in cervical myelopathy. Postural scintimyelography is a refinement of scintimyelography and in combination with Queckenstedt's test proved to be an exellent screening method for possible space occupying lesions in the cervical subarachnoid space. The results in a material of 96 patients are reported. Agreement between both methods is of high clinical significance.


Videodensitometry of cerebrospinal fluid pulsations. Roentgen videodensitometry (RVD) during myelography is described in 13 patients with a variety of intraspinal lesions. Abnormalities in the RVD tracing are discussed and correlated with the type of spinal canal obstruction. Intramedullary, intradural and extradural processes may be differentiated by RVD.


Metrizamide, the new water-soluble non-ionic contrast medium for myelography. Clinical experience. A survey is made of the development of water-soluble contrast media and of the experimental and preliminary clinical investigations of the new water-soluble contrast medium metrizamide. The experiences with the medium indicate that its use in the subarachnoid space is safe and reliable.


Perfluoro carbon bromides as contrast media in radiography of the central nervous system. A preliminary experimental report. A preliminary investigation of the properties of perfluorooctyl bromide in the subarachnoid space revealed this compound to have suitable attenuation capacity and advantageous physical and biologic properties for demonstration of the brain and spinal cord. No acute or chronic neurotoxicity was encountered. Microscopy demonstrated evidence of chronic inflammatory changes subsequent to long term exposure to the compound. It was concluded that the medium was superior to iodophendylate in several respects and that it may provide an alternative to water-soluble contrast media. Perfluorohexyl bromide proved unsuitable for neuroradiologic use because of formation of excessive amounts of non-absorbable gas, leading to high subarachnoid pressure and severe neurologic deficits.


Advances in ventriculography and myelography with the non-ionic constrast medium metrizamide. Laboratory experiments in animals have shown that the non-ionic constrast medium metrizamide is well tolerated by the central nervous system. The preliminary clinical experiences have confirmed this exceptional tolerance. The extremely weak epileptogenic action of this compound represents the most important progress.


Metrizamide in radiography of the central nervous system. A preliminary report. Metrizamide seems to be a suitable contrast medium for ventriculography and myelography. This contrast medium is less toxic and far less epileptogenic and spasmogenic than the other water-soluble contrast media in current use. No serious complications were noted in the present series. Transient changes in EEG were recorded, mainly with cervical myelographies. However, it is too early to evaluate the long-term side effects.


Preliminary experiences with the EMI scanner. Preliminary experiences of computed tomography of the head are reported in a material of 213 patients. The majority of examined lesions includes intracranial tumours and cerebrovascular lesions.


First results with an EMI scanner.

First results with an EMI scanner. The results of computer assisted tomography (80 x 80 matrix) of 600 patients are presented.


Correlation between the appearances of the ventricular system at carotid angiography and computer assisted axial tomography. Computer assisted axial tomography and the venous phase of carotid angiography provide complementary assessments of the ventricular and paraventricular structures. The computer assisted tomography more readily defines the ventricular system, but can prove weak in differentiating paraventricular gliomas, solitary metastasis and infarcts. Angiography plays a major role in differentiating between these three conditions, but is poor in accurately localizing and identifying intra- and paraventricular hemorrhage which is characteristically and expeditiously uncovered by C.T. scanning.


Radiology of suprasellar ectopic pinealoma. The radiographic and clinical findings in twenty-three recently reported cases of suprasellar ectopic pinealoma are reviewed and three previously unreported cases are described. It is concluded that suprasellar calcification is extremely rare; enlargement of the sella turcica was present in approximately 20 to 30 per cent, none demonstrated elevation of the anterior cerebral artery. Pneumography may show a smooth or irregular filling defect in the anterior part of the third ventricle. In one of the new cases there was an isotope uptake in the suprasellar region. The differential diagnosis and other clinical data are also discussed.


Accuracy of scanning, angiography and encephalography in posterior fossa tumors. A material of 120 cases with posterior fossa tumors examined with scanning, vertebral angiography and encephalography was reviewed to determine the value of the information and accuracy these methods provide. Encephalography was the most accurate procedure (98% positive) compared to angiography (81%) and scanning (44%).


Technique for internal auditory cisternography. A technique for internal auditory cisternography is described. A small volume of Iophendylate is used and coronal hypocycloid tomographic sections of the dependent internal auditory canal and adjacent cerebellopontine angle cistern performed.


Quantitative analysis of the pituitary fossa and gland in Cushing's disease. With tomography and encephalography of 78 cases with primary Cushing's disease, changes in the appearance of the pituitary gland and fossa have been demonstrated in greater numbers than previously reported. One or more direct or indirect signs of enlargement of the pituitary gland either by hyperplasia, microadenoma, or adenoma occurred in 60 per cent. Most of these cases were unproved as to cell type because they were given proton beam therapy. There were more cases of pituitary enlargement in those with adrenalectomy in comparison to non-adrenalectomy cases and in those with pigmentation in comparison to non-pigmented cases. The high incidence of pituitary gland enlargement in the different phases of this disease supports the importance of this gland in the pathogenesis of primary Cushing's disease. Limitation in correlation of the pituitary fossa to size of the pituitary gland when the latter enlarges only slightly has been discussed.


Developmental defects of the cerebellum. A radiologic and anatomic investigation. A classification of cerebellar malformations is proposed, based on more than fifty cases with varying degrees of abnormality, where comparative anatomic and radiologic analysis was carried out. The anatomic information was collected at surgery or autopsy and the radiologic evidence was derived from encephalography or ventriculography and vertebral angiography. The malformations were classified in four groups in order of gravity, with regard to the morphologic and etiologic features: (1) total or sub-total agenesia of the cerebellar structures (vermis and hemispheres); (2) almost complete agenesia of the vermis associated with the Dandy-Walker malformation; (3) almost complete agenesia of the vermis without the Dandy-Walker malformation; (4) partial defects of the vermis, which could be separated into four subdivisions.


Prolactin-secreting pituitary microadenomas. Prolactin-secreting pituitary microadenomas have been surgically selectively removed in 20 patients presenting an amenorrhea-galactorrhea syndrome, hyperprolactinemia and radiological modifications of the sella turcica. In these patients, the sella was of normal size but hypocyloidal tomography evidenced subtle changes characterizing pituitary microadenomas. The surgical intervention has been based on the twin criteria of hyperprolactinemia and sellar findings.


Iatrogenic intracranial aneurysms.

Iatrogenic intracranial aneurysms. Intracranial aneurysms following head trauma are not uncommon. Few cases of iatrogenic intracranial aneurysm have been reported however. This type of lesion is illustrated by three cases. If these lesions have the same poor prognosis as that proposed for the usual traumatic aneurysm, their early diagnosis and treatment is important.


Accessory meningeal artery.

Accessory meningeal artery. The accessory meningeal artery, originating from either the internal maxillary or middle meningeal artery, is predominantly an extracranial vessel. A minor branch of the artery enters the skull through the foramen ovale to supply the dura of the temporal fossa as well as the Gasserian ganglion. This intracranial branch may be demonstrated on angiography and may serve as a collateral between the external and the internal carotid arteries, with or without a complex intervening vascular network. The accessory meningeal artery may be identified as the arterial pedicle responsible for blood supply to meningiomas of the temporal fossa and possibly even to tumors of the Gasserian ganglion.


Radiologic evaluation of the function of the cervical spine. Cervical functional examinations initiated in 1965 suggested the need of a correlated, multifactorial expression of the cervical sagittal movement function, allowing comparative evaluations of physiologic-diagnostic significance. Technical aspects of the successive steps of investigation, together with the final morphologic-functional formula are described. The significance of the normal and abnormal functional patterns, identified in a selected series of 300 cases, is finally discussed.


Fusiform aneurysmal dilatation of pericallosal artery. A sign of lipoma of corpus callosum. Two cases of lipoma of the corpus callosum with aneurysmal dilatation of the pericallosal artery are reported. This appears to be a reliable sign of lipoma of the corpus callosum. Our cases and the literature reveal that 19 of 22 cases with illustrations of carotid angiography have demonstrated such an aneurysmal dilatation.


Early venous filling and controlled hyperventilation in cerebral angiography. Controlled hyperventilation is a useful adjunct to angiography of tumours and inflammatory disease, giving increased demonstration of normal penetrating arteries and abnormal vessels. This shunting effect commonly causes early filling of the deep ependymal veins which should not be mistaken for early venous return due to tumour. While the effect could theoretically mask early venous flow secondary to tumour, this has not been our experience.


Sylvian triangle. Angiographic evaluation of its appearance in normal and pathologic conditions. Description of a method for determining the size of the Sylvian triangle on lateral films. A cranio-insular index is constructed which allows differentiation between normal cases, paracapsular and subdural hematomas.


[Effects of intrathoracic pressure on cerebral circulation in the monkey as a function of body posture, sitting or standing]. Sixty-four measurements of CBF were performed in 8 monkeys in supine and in sitting position under various conditions of ventilation. The position of the body does not normally influence the response of brain circulation to CO2. The response is not different from normal with high insufflation pressure in supine position and the capacities to autoregulation seem to be maintained. In vertical position however there are no longer any significant relations between CBF and PaCO2, and the loss of autoregulation seems to occur earlier and to be more marked. Correlation of these findings to neuroradiologic data indicates that the cerebral venous drainage through the meningospinal plexus is enhanced in vertical position and that during intrathoracic hyperpressure the cerebral venous blood is retained in the jugular vein and meningospinal plexus, thus decreasing the cerebral venous pressure increase and maintaining a perfusion of brain tissue compatible with the survival of the animal.


Phlebographic appearance and pathogenesis of venous malformations in the orbit. A retrospective clinical and radiographic analysis of 20 patients with orbital venous malformations was performed. The malformations were fed by other veins rather than arteries. They may nearly always be demonstrated by orbital or jugular phlebography, though occasionally they will fill only following direct puncture. There appeared to be varied etiologies. A few were related to trauma or to other causes of increased intraorbital pressure. There is an association with periorbital hemangiomas suggesting developmental or congenital origin.


Cerebral arterial spasm. II. Etiology and treatment of experimental cerebral vasospasm. Delayed cerebral vasospams is caused by excessive accumulation of dopamine-beta-hydroxylase (DBH) and noradrenaline in cerebral vessel walls. This study demonstrates the mechanisms of delayed spasm, particularly the role of red blood cell components, and the successful relief of delayed cerebral vasospasm. Spasmogenic substances which contained a heme component, such as methemoglobin, methemalbumin and catalase enhanced DBH activity in human serum as measured by a one step chemical spectrophotometric assay. The concentration which gave the highest DBH activity caused the maximum constriction of the basilar artery, when the substances were applied topically. Among components of red cells, methemoglobin, methemalbumin, catalase and nicotinamid adenin dinucleotide (NADH) caused constriction of basilar artery in cats, when applied topically, whereas hematin, hemin and bilirubin caused no significant spasm. An oxyhemoglobin solution obtained by mixture with methemoglobin and ascorbic acid produced no significant vascular spasm either. Relief of delayed cerebral vasospasm was obtained with topical application of specific alpha adrenergic blocking drug such as phenoxybenzamine, specific inhibitors of DBH such as fusaric acid, o-phenanthroline and alphaalpha' dipyridyl beta2 adrenergic stimulants such as salbutamol, and a phosphodiesterase inhibitor, ascorbic acid.


Dedifferentiation and invasive growth of an eosinophil pituitary adenoma. This report concerns the course of an eosinophil pituitary adenoma in an acromegalic female (16 years of age when first symptoms appeared) over a period of ten years. The case was complicated by craniocerebral trauma and CSF rhinorrhoea. After several operations, dedifferentiation, and invasive growth into the orbit and the petrous bone were observed with walling in of the VIIIth cranial nerve. The possible causes of the dedifferentiation of the adenoma are discussed.


The mode of action of the nicotinic cholinergic receptor protein in the postsynaptic membrane. The functional properties of the cholinergic (nicotinic) receptor protein were discussed. on the basis of in vitro studies of membrane fragments of Electrophorus and Torpedo electric tissues. The membrane fragments of both are excitable in vitro. In contrast with the membrane fragments of Electrophorus, however, those of Torpedo give dose-response curves of in vitro excitation that shift towards higher concentration of the agonists by one to two orders of magnitude compared with the actual binding curves of agonists to the receptor sites. Moreover, they exhibit the phenomenon of pharmacological desensitization in a completely cell-free system. It was suggested that the discrepancy between the dose-response and the binding curves is related to the desensitization. The mechanism and the molecular events of desensitization are discussed. Desensitization is a reflection of a conformational change of the receptor protein during the excitation caused by agonists. This conformational change causes desensitization in subsynaptic membranes, through amplification. The regulatory properties of the cholinergic receptor protein similar to those of allosteric enzymes were discussed


Cytochrome oxidase and urate oxidase as intracellular O2 indicators in studies of O2 gradients during hypoxia in liver. Results obtained from a new approach of investigating O2 distribution in intact perfused liver under hypoxic conditions are presented. Based on the signals observed by organ absorbance spectrophotometry from two compartments with oxidases of markedly different O2 sensitivity, the mitochondria and the peroxisomes, a distribution between high O2 and zero O2 zones is postulated, an intermediate border zone of O2 concentrations between the K0,5 (O2) values being virtually absent (steep intercellular O2 gradients).


Transient metabolic and vascular volume changes following rapid blood pressure alterations which precede the autoregulatory vasodilation of cerebrocortical vessels. It has been shown by surface fluoro-reflectometry that stepwise decrease of arterial blood pressure causes a biphasic cerebrocortical vascular volume response. After the arterial blood pressure decrease the vascular volume first decreased and later increased. In both parts of the biphasic reflectance change, the cerebrocortical NAD-NADH redox state shifted considerably towards reduction and there was no reoxidation after the onset of cortical vasodilatation. Since a very rapid NADH reduction occurred during the first 30 secs. of the arterial hypotension in parallel with the vascular volume decrease, it is suggested that in the transient phase of arterial hypotension cerebral hypoxia may occur. Furthermore it is suggested that anaerobic tissue metabolites or some unknown NAD-NADH dependent process might dilate the cerebrocortical arterial network during the autoregulatory adjustment of CBF. The participation of the sympathetico-adrenal system in transient brain hypoxia caused by bleeding is a possibility since both the early vasoconstriction and the steep NADH reduction were prevented by the administration of phenoxybenzamine (1 mg/kg) before bleeding.


Prognostic significance of morphologic and cytochemical markers in adult acute leukemia. Survival times in 100 cases of acute leukemia (74 granulocytic, 14 lymphocytic, and 12 undifferentiated) were correlated with classic morphologic and cytochemical criteria. The 14 patients who had lymphocytic leukemia had significantly longer survival compared with the two other groups. Undifferentiated leukemias had a shorter survival time than granulocytic leukemias. Several subclasses of granulocytic leukemias were formed according to the presence or absence of Auer rods and the percentage of peroxidase-positive blasts. Neither of these two features significantly influenced the survival of these patients. In the lymphocytic leukemia group, PAS-positive and negative leukemias had similar survival expectancies. It is concluded that division into lymphocytic and nonlymphocytic leukemias is still helpful in predicting survival times of patients who have acute leukemia, but that further subclassification of these groups based on the presence or absence of Auer rods and the percentages of peroxidase-positive blasts is of no additional benefit.


Growth hormone deficiency, brain development, and intelligence. Twenty-nine patients with growth hormone (GH) deficiency were selected according to the following criteria: no evidence of reversible GH deficiency, onset of growth retardation in early childhood, and no evidence of pituitary tumors or other direct pituitary trauma. Fourteen patients had evidence of multiple hormone deficiencies, 14 had isolated GH deficiency, and one patient questionable isolated GH deficiency. Psychometric testing showed a normal IQ distribution. The GH deficiency was not associated with deficiencies in specific mental abilities. Likewise, GH treatment in later childhood and adolescence did not seem to influence intelligence. Patients with multiple hormone deficiencies had somewhat lower IQs than patients with isolated GH deficiency when socioeconomic status was controlled. We conclude that GH deficiency itself does not seem to affect human brain development and intelligence.


Superoxide generation by human monocytes and macrophages. Intracellular and extracellular superoxide (O2.-) generation by human monocytes and macrophages was quantitated by the nitroblue tetrazolium (NBT) reduction method. Human monocytes reduced 4.4 +/- 0.9 nmoles/10(6) cells/15 minutes with an increase to 12.4 +/- 1.3 during phagocytosis of zymosan. Based on inhibition by superoxide dismutase, superoxide generation of these cells was 1.8 +/- 0.9 nmoles in the resting state and 16.8 +/- 2.8 nmoles with zymosan phagocytosis. Human macrophages obtained by thoracentesis had comparable levels of NBT reduction and O2.-generation. Monocytes from a patient with chronic granulomatous disease demonstrated no increment in O2.-production during phagocytosis. Thus, human monocytes and macrophages appear capable of generating substantial amounts of O2.-during phagocytosis which may play an important role in bactericidal and other cell functions.


Uridine monophosphate kinase polymorphism in two Venezuelan populations. A study of the uridine monophosphate kinase (UMPK) electrophoretic patterns in Venezuelan individuals from the mestizo population of Caracas and from the Warao Indians of the Nabasanuka village in the Delta of the Orinoco River are reported. Among the mestizo population, the frequency of the UMPK1, UMPK2, and UMPK3 alleles was .979, .020, and .001, respectively. A higher frequency of the UMPK3 gene was seen in the highly inbred Warao Indians than any other population studied to date.


Consensual reactions to anterior chamber paracentesis in the rabbit. A consensual reaction was consistently obtained in rabbits subjected to unilateral paracentesis. On fluorescein angiography of the anterior segment, we observed that this consensual reaction consisted of a slight to moderate dye extravasation from the ciliary processes into the aqueous. Aspirin usually, but not always, inhibited this reaction. Retrobulbar injection of anesthesia to the injured eye always blocked the consensual response, as did phenoxybenzamine injected intravenously before the paracentesis. We concluded that consensual responses are more efficiently inhibited by nerve-blocking agents than by prostaglandin inhibitors. These results suggest that the interocular pathway mediating consensual responses is probably neural and not caused by prostaglandins released into the general circulation.


Orbital fibrous histiocytoma in an infant. A case of fibrous histiocytoma of the orbit occurred in a 1-year-old infant. The tumor was excised, and no recurrence has been evident.


The lung of the copper-deficient rat. A model for developmental pulmonary emphysema. Based on the hypothesis that cross-linked elastin is critical for normal lung structure, lung tissue from copper-deficient rats was studied. Copper deficiency was induced in the second generation by feeding dams a milk-based diet low in copper (less than 1 ppm) during gestation and lactation. The weanlings were fed the same diet until they showed severe signs of deficiency between 6 and 10 weeks of age. Controls animals received the basal diet supplemented with 10 ppm copper. Liver cytochrome oxidase activity, which served as the chief index of deficiency, decreased from a normal level of approximately 80 to 15 mumole/min/g. The lungs of the deficient animals contained 17% less elastin and had 35% larger alveolar spaces (34.7 vs 47.7 intercepts), as determined by the mean alveolar intercept method. The ultrastructure of elastin in the bronchi, arterioles, and alveolar ducts had a "washed out" appearance. To determine the reversibility of the pathology, deficient animals, 5 to 10 weeks of age, were repleted by feeding a copper-supplemented diet for 1, 2, and 3 months. During this period growth resumed, anemia disappeared, and liver cytochrome oxidase returned to normal. There was no improvement in lung structure with regard to alveolar size (28.4 intercepts compared with 43.6 in controls and 35.1 in deficient littermates killed at the start of repletion). The ultrastructure and electron density of pulmonary elastin was restored to near normal. The lung of the copper-deficient rat is proposed as a model for developmental pulmonary emphysema.


The effect of phorbol myristate acetate on the metabolism and ultrastructure of human alveolar macrophages. In the present investigation we examined the influence of the surface-active agent phorbol myristate acetate (PMA) and opsonized heat-killed bacteria (HKB) on oxygen consumption, superoxide release, and glucose oxidation of human alveolar macrophages (AM). Both PMA and HKB produced a surge in oxygen consumption, superoxide release, and oxidation of 1-14C-glucose and 6-14C-glucose by human AM. Examination of AM by electron microscopy following stimulation by these two agents demonstrated membrane ruffling, loss of microvilli, and increased vacuolization in PMA-treated cells and phagocytic vacuoles containing bacteria in HKB-treated cells. The vacuolization produced by PMA-treated AM was much less striking  than the vacuolization produced in PMA-treated leukocytes. The similarity in the metabolic and some of the physical responses of AM stimulated by PMA and HKB suggest that PMA may be a useful agent for evaluating cell-membrane-related events of phagocytosis in AM.


Expression of herpesvirus in adherent cells derived from bone marrow of latently infected guinea pigs. The role of bone marrow adherent cells in the latency of guinea pig herpes-like virus (GPHLV) was explored. Cultures of macrophage-enriched adherent cells derived from infected guinea pigs were examined for evidence of latent GPHLV infection. Expression of the virus was detected in these cultures 9 to 10 days after in vitro cultivation. Increasing virus infectivity titers as well as light and electron microscopic evidence of virion assembly in macrophages and fibroblasts were demonstrated. Infections virus was detected in the bone marrow adherent cells that had attached for 30 or 120 minutes but only following reverse cocultivation. The data showed not only that the bone marrow adherent cells were susceptible to GPHLV in vitro but also that GPHLV was harbored by the macrophage-enriched bone marrow population in vivo in latently infected guinea pigs.


The opiate receptor and morphine-like peptides in the brain. Opiate receptors--neuronal membrane proteins that have been identified by the direct binding to membranes of radioactive opiates--have been shown to mediate the pharmacological effects of opiate drugs. Examination of brain extracts for substances that mimic effects of opiates on the opiate receptor permitted identification of the enkephalins, two peptides, each containing five amino acids, which are the normal substrates for the opiate receptor. Enkephalins are contained in specific neurons localized to areas of the brain enriched in opiate receptors. Enkephalin-containing neurons and opiate receptors are concentrated in portions of the brain that mediate pain perception, emotional behavior, and other functions altered by opiates. beta-Endorphin, an opiate-like peptide containing 31 amino acids, is localized to the pituitary gland from which it can be released into the circulation to act presumably at peripheral target organs. In the brain beta-endorphin is concentrated in the hypothalamus.


Brief versus standard hospitalization: the differential costs. The authors compared the use of inpatient and day care services, number of readmissions, use of special services, use of drugs, costs to family and community, and differential dollar costs of three treatment approaches--brief hospitalization followed by day care, brief hospitalization followed by outpatient care, and standard hospitalization. They found that, among patients who had families willing to care for them, brief hospitalization followed by either day or outpatient care was less expensive in terms of hospital costs and costs to the family than standard hospitalization.


The paradoxical underutilization of partial hospitalization. Partial hospitalization continues to be underutilized even though its clinical effectiveness for a variety of psychiatric patients has been demonstrated. The authors investigated the potential economic advantage of partial hospitalization by comparing matched groups of day hospital patients and inpatients who had comparable symptoms and prognoses on admission. They present one-year follow-up data documenting the comparability of the study groups on clinical outcome measures and the cost advantages favorable the partial hospitalization group. They discuss possible causes of the paradoxical underutilization of the clinically effective and lower-cost partial hospitalization, which include institutional factors, patients' clinical characteristics, family resistance, and clinician bias.


Electron-microscopic study of pseudo-exfoliation of the lens capsule. The lens, iris, and ciliary body from the eyes of patients with glaucoma with cataract were examined via electron microscopy, and the following results obtained. 1. Zonular bundles were encrusted with accumulations of pseudoexfoliative (PE) materials and fragmented zonular fibrils, especially on their lateral aspects. 2. PE materials ranged from 300 to 400 A in width, consisting of several subunits about 100 A wide. 3. Pigment epithelial cells of the iris and nonpigmented epithelial cells of the ciliary process occasionally contained many phagosomes engulfing pigment granules. 4. Near the ciliary process, degenerating zonular bundles were mostly replaced with PE materials and fragmented zonular fibrils, and a gradual transition was observed between them. From the above findings, it is probable that PE material might be mainly composed of degenerated zonular fibrils, which are presumed to be digested with lysosomal enzymes.


The public health significance of mycotoxins. Diseases of man and animals associated with foodstuffs contaminated by molds have been known for centuries, but only recently have mycotoxicoses been recognized as a significant hazard to man. A detailed review of the literature involving the major mycotoxin-producing fungi is presented. Because of their hazards to both human and animal health, increasing concern has developed regarding the carcinogenic activity of aflatoxins and other mycotoxins. The symptoms of mycotoxicoses in man and experimental animals is reviewed and certain specific case histories reported. The industry is urged to implement all available techniques in order to minimize the contamination of foods by mycotoxins.


Cytomegalovirus retinitis: two cases occurring after renal transplantation. Cytomegalovirus (CMV) retinitis is reported in 2 patients who received immunosuppressive medication after cadaveric renal transplantation. The clinical features of this disease as well as the differential diagnosis are presented. Reduction in the amount of immunosuppressive drugs given postoperatively is suggested as the definitive treatment of this disease. Available specific antiviral chemotherapeutic agents were ineffective against CMV retinitis. This finding may be modified at a later date, with the introduction and use of new effective and safe antiviral drugs.


[Frequency of viral infections (especially with herpes virus) during Darier's and Hailey-Hailey's diseases (author's transl)]. The fragility of epidermal cells in Hailey-Hailey's disease end even more in Darier's disease seems to favour the development of viral infections. The authors present six cases of herpes virus infections in three siblings with an intermediate form between Darier's disease and Hailey-Hailey's disease, both having a tendency to recur. One of the flares demonstrated signs of Kaposi and Juliusberg's pustulosis vacciniformis which is often found in the literature as a complication of Darier's disease. The authors discuss the frequency of the recurrences, which could be due to different viruses. Herpetic virus is often found, less frequently vaccinial virus, but sometimes also Coxsackie A 16 virus.


[Double-blind comparative study of trimethroprim-sulphacetamide-polymyxin b and gentamicin in the treatment of otorrhoea (author's transl)]. A double-blind comparative study of the use of Garamycine (gentamicin) ear drops and TSP (trimethroprim-sulphacetamide-polomyxin B)  ear drops in the treatment of 100 cases of otorrhoea due to external otitis, a recurrent otitis media accompanied by perforation of the drum, an infection of the mastoid cavity or a postoperative infection, provided evidence of the effectiveness of both medications. TSP ear drops gave positive results in 42 out of 50 cases treated, whilst for gentamicin aural solution results were positive for 46 out of 50 cases. Gentamicin gave a successful result in 5 of the 8 failures with TSP, whilst TSP cured the four original failures with gentamicin. There were no signs of ototoxicity, of excessive fungal proliferation nor of any local sensitivity to the ear drops. It would seem that these aural preparations are complementary, capable of resulting in the disappearance of the majority of bacterial agents responsible for pathogenic otorrhoea.


[Partially hormone dependent bilateral malignant polyadenomatous hyperplasia]. Bilateral malignant polyadenomas of adrenals seem to be rare. They involve either a double primary adrenal tumour, or a metastasis of a controlateral tumour. In the observation reported here, the evolution is characterized by periods of central stimulation with partial autonomy, and periods of tumoral autonomy with central inertness. Pathological findings consist of coexistence of various histological aspects (atrophy, hyperplasia, benign adenoma, carcinoma) suggesting the successive steps of a very particuliar adrenal carcinogenesis. Such findings allow to discuss the following physiopathological mechanism: a stimulation by ACTH might have resulted initially in a preneoplasic hyperplasia, then in an incompletely autonomous tumour, and finally in an autonomous tumour. Accordingly, from a practical and therapeutic point of view, it would be suitable, after removal of an adrenal tumour, especially of one with demonstrated ACTH-dependance, to suppress totally endogenous ACTH by cortisol or cortisone therapy, in order to reduce the occurence of corticotropin stimulation of possibly remaining malignant adrenal cells.


Neurilemoma of the jugular foramen. Transmastoid removal. The authors present two cases in which the palsies of the IX, X, XI and XII nerves heralded the presence of a neurilemoma within the jugular foramen. Temporary bone polytomography, retrograde jugular venography, and pantopaque posterior fossa myelography allowed the nature and extent of the tumor to be predicted accurately prior to the operation. The tumors were completely removed using a transmastoid-extended facial recess approach without labyrinthectomy, supplemented in one case by exploration of the upper cervical portion of the internal jugular vein. This technique has avoided the complications which attend removal by way of the posterior fossa. We feel that transmastoid removal merits more extensive clinical trial.


[Pituitary adenoma with alpha, 17-39 ACTH and beta, MSH cells, without hypercorticism (author's transl)]. In one case of pituitary basophil adenoma, the majority of the cells contains numerous small granulations which, by using immunohistoenzymatic technique, react with alpha, 17-39 ACTH and beta, MSH antisera. The reaction with anti beta, 1-24 ACTH is negative. The radioimmunoassay of ACTH reveals an increase of plasmatic ACTH, but the cortisol cycle stays low. This data suggests that this functional adenoma releases a product of which has an immunological relationship with ACTH, without biological activity.


[Hyponatremia, a frequent, and at times a major and indicative sign in anterior pituitary insufficiency]. Thirty five patients with hypopituitarism have been studied. Nine had a mild hyponatremia (126-137 mEq/l) and five a profound hyponatremia (105-124 mEq/l). None of them had hemodilution (plasma protein, potassium were normal). Hyponatremia was present even in the absence of deficit in GH and/or TSH, and/or FSH-LH. All the pateints had a complete ACTH deficiency. There was a negative correlation between age and natremia (r = -0,39; p0.02). 2/5 patients with severe hyponatremia had mental confusion. These two patients had a complete TSH deficiency. Natremia increased with cortisone therapy in all cases but reached the normal range only in one case. Mental confusion disappeared and natremia became normal in all patients with therapy with cortisone and thyroid hormones.


Study of 100 patients with bilateral sensorineural hearing loss for lipid abnormalities. One hundred persons were selected from our audiology records who showed at least 20 dB bilateral sensorineural hearing loss. The number 100 was picked for an adequate statistical analysis. The patients were taken from the chart files which are filed in chronological order. Letters were mailed with instructions for fasting 14 hours prior to testing except for the consumption of water. On presentation, a history was taken for diabetes, height and weight, and the blood pressure was recorded. The blood samples were drawn in clot tubes and taken to a commercial laboratory for overnight refrigeration and testing of the serum. Testing included observation of serum for massive chylomicronemia, triglycerides, cholesterol and lipoprotein electrophoresis (by the cellulose acetate method). Lipoprotein testing results showed 12 patients with Type II A or II B and 8 patients with Type IV abnormalities. No Types I, III or V were found. These 20 patients represent a lower number of hyperlipoproteinemic patients than would be expected in the general population.


Synovial sarcoma of the hypopharynx.

Synovial sarcoma of the hypopharynx. Primary synovial sarcoma of the hypopharynx is an extremely rare neoplasm. Only 16 cases could be traced in the literature so far. An additional case, in a 29-year-old male patient, is presented. Progressive disturbance of deglutition and speech, chocking sensation, and mild respiratory distress were his original complaints. Our case differs from those previously reported in that a) the tumor was pedunculated, and the site of its attachment could be very well-defined, and b) unusual intratumoral destructive hemorrhage turned the case into an emergency. The pedicle was radically excised together with part of the grossly uninvolved underlying tissue, and the tumor was removed in toto. The histological findings are described. The patient is under observation. No recurrence has been noticed 11 months after surgery, and there are no signs of metastasis. The origin of the tumor and its treatment are discussed.


[Inorganic polyphosphate metabolism in Staphylococcus aureus and the action on it of antibiotics]. The culure of Staph. aureus in the exponential growth phase contained 14-18 mg/g of dry orthophosphate biomass and 18-22 mg/g of dry acid insoluble polyphosphate biomass. The extracellular extract of the culture had a phosphohydrolase activity with respect to high molecular polyphosphates, tripolyphosphate and pyrophosphate. Penicillin and bacitracin which inhibited the biosynthesis of the cell wall had no effect on the content of polyphosphates and the phosphohydrolase activity of Staph. aureus. Heliomycin which inhibited the biosynthesis of RNA increased the content of polyphosphates by 1.5 times and decreased the content of ATP by 30 per cent in the cells of Staph. aureus.


Practical method for detecting poliovirus in anaerobic digester sludge. A method has been developed which permits reliable detection of low numbers of poliovirus infective units in anaerobic digester sludge and dewatered composted sludge.


Primary versus secondary nerve repair: a review of the literature. The pros and cons of primary and secondary neurorrhaphy are discussed on the basis of a study of the literature. The impression is gained that recently, many surgeons have come to prefer secondary restoration of the continuity of a severed peripheral nerve. However, the longer the delay, the more serious the problems and the worse the prognosis. The authors conclude therefore that repair of the nerve in the earliest possible stage should always be attempted. Provided the relevant conditions are fulfilled, primary neurorrhaphy should be seriously considered; the prognosis of primary repair is good, especially in children. If primary neurorrhaphy is contra-indicated, early secondary repair is to be preferred to further delay.


Ultrastructure and x-ray microanalysis of siderosome. Comparative ultrastructural examination and energy-dispersive electron probe X-ray microanalysis were performed on a long standing skin lesion of hemosiderotic histiocytoma. Iron-containing fine particles present in siderosome were the main element of interest. Qualitative study of spectra over the siderosomes clearly demonstrated the characteristic X-ray energy emitted by iron, whereas spectra obtained from adjacent cytoplasm revealed minimal iron peak. On quantitative evaluation of the spectra yielded from various siderosomes, iron counts intensity was found to be proportionally increased with increment in amount and electron opacity of the siderosomal inclusion. Accounts on chemical nature of the siderosomal inclusion and on the presence of lipid residue in cytoplasms were noted.


[Multivesicular vacuolization in malignant histiocytoma of the skin (author's transl]. A 47-year-old patient developed several rapidly growing tumours in his right leg. Histiologic and electron microscopic examination revealed morphological characteristics suggesting the histiocytic origin of the tumour cells. An important finding was the striking number of pleomorphic membraneous inclusions and large vacuoles containing numerous small vesicles. These cytoplasmic inclusions are interpreted as transformed multivesicular bodies. Their formation may be an expression of an increased and bizarre membraneous activity of a malignant cell race. Radiotherapy resulted in a remarkable rapid and complete remission.


"Total" therapy for small cell carcinoma of the lung. Seventy-one consecutive patients with small cell carcinoma of the lung were treated with an integrated approach between November, 1974, and May, 1977. The regimen included radiotherapy to the primary site, relatively brief (6 to 12 weeks) although intensive chemotherapy, and prophylactic cranial irradiation. Complete responses were achieved in 75% and 40% of patients with limited and extensive disease, respectively. Modest prolongation of survival (median, 10 months) was realized by patients with extensive disease, but prolonged relapse-free survival was not observed. In contrast, one-half of the patients with limited disease who achieved a complete response have remained clinically free of disease without further treatment for a mean of 18 months (range, 6 to 33 months). Since all relapses to date have been noted within the first year following cessation of treatment, this experience suggests there may be a potential for cure in those survivors who are now relapse free for intervals exceeding two years.


[Ultrastructure and clinicopathological aspects of mucoepidermoid tumour of the salivary glands (author's transl)]. The mucoepidermoid tumor of the salivary glands is relatively rare; its ultrastructure is analysed on the basis of four cases. Three secretory tumour cells can be differentiated: one tumour cell with mucous secretion and two tumour cells with abortive secretion which are different in structure. The dysregulated and abortive secretion is due to a defective functional differentiation and defective innervation of tumour cells. Further characteristic tumour cells without secretion are: epidermoid cells, myoepithelial cells and clear cells. The variety of the tumour cells in the tissue is due to the broad cytogenic prospection of acinar and tubular epithelium of the salivary glands. Recommendations are given for practical electron-microscopic diagnosis by means of typical leading elements of the structure. The author tries to assess the gravity of the mucoepidermoid tumour by ultramicroscopic findings. The tumour is classified as potentially malignant. Therapeutic aspects are discussed dependent upon this grading.


Quantitative nuclear-cytoplasmic ratios in human hepatomas. This report presents quantitative data on nuclear-cytoplasmic (N/C) ratios of human hepatomas, normal liver specimens, and cirrhotic liver specimens. The N/C ratios were determined by point-counting, using an intraocular microscopic grid as the source of the points. The N/C ratios of hepatoma cells were chiefly greater than 0.21, while those of normal and cirrhotic liver cells were chiefly less than 0.21. The N/C ratios of hepatoma cells had a variance significantly greater than those of normal and cirrhotic cells. Quantitative N/C ratios can yield information useful in characterizing cell types for diagnostic purposes.


Metastatic tumors of the endocardium: report of three cases. Because of their apparent rarity and the tendency of clinicians to lump indicative signs and symptoms under the heading of metastatic disease, metastatic tumors of the endocardium are seldom mentioned in the literature, in the three cases presented herein, endocardial metastases were evident at autopsy. In one case of malignant melanoma, clinical evidence for endocardial involvement was present in life. This article also presents a case of endocardial involvement by Wilms' tumor and a case of endocardial involvement by hypernephroma with pulmonry tumor emboli.


Cystosarcoma phyllodes in young women.

Cystosarcoma phyllodes in young women. A total of eight cases of cystosarcoma phyllodes in women under 25 years of age were reported to the Swedish Cancer Registry during a ten-year period (1960 to 1969). Only one of these tumors was malignant. It was successfully treated with simple mastectomy and postoperative radiation therapy. Of the remaining seven cases, six were treated with local excision of the lesion and one with simple mastectomy. In no case, did the tumor recur during a follow-up of an average of ten years. Local excision seems to be sufficient for the benign form of the tumor, whereas simple mastectomy is indicated for the malignant form. Radical mastectomy is indicated only of the tumor has invaded the pectoral fascia. The tumor rarely metastasizes to the axillary lymph nodes.


Isolation of a variant of BK virus with altered restriction endonuclease pattern. A papovavirus was isolated from the urine of a 24 year old female who underwent bone marrow transplantation after a relapse of acute myeloid leukemia. The virus (JL) resembles BK virus in its antigenic and growth properties, but has a different restriction endonuclease pattern after digestion with the restriction endonucleases Eco R1 and R Hind II + III.


Stability of neurotropic mouse hepatitis virus (JHM strain) during chronic infection of neuroblastoma cells. A line of mouse neuroblastoma cells which was chronically infected with the neurotropic strain (JHM) of MHV, a member of the coronavirus group, was established. These cells, designated NJ, exhibited typical MHV cytopathic effects (CPE) at all passage levels along with the continual production of infectious virus. Most cells were positive for viral antigen by immunoflourescence. Viral particles consistent with the morphology of MHV were found by electron microscopy. The uninfected neuroblastoma cell line did not contain a detectable population of cells resistant to JHM, and persistence did not elicit the production of interferon. No plaque morphology or temperature sensitive mutants were selected for in the NJ culture, and we were unable to detect the presence of either a defective or defective interfering virus population. The addition of low concentration antiviral antibody modulated the infection to a carrier culture with viral antigen in the cytoplasm of the cells, but no infectious virus was produced, and the cells lacked both surface viral antigen and CPE. Possible mechanisms of viral persistence in vitro are discussed.


Interferon production and activity in mouse neuroblastoma cells. Mouse neuroblastoma cells are capable of synthesizing interferon after induction with virus or polynucleotides. Differentiation of neuroblastoma cells depresses interferon synthesis without an apparent effect on sensitivity to interferon.


[Value of electroencephalographic findings in the prognosis of Lennox-Gastaut syndrome]. The electroencephalographic findings observed in twenty-nine patients with Lennox-Gastaut syndrome were related with prognosis of the epileptic manifestations. The presence of slow spike-wave complexes with preponderance of spike over the slow wave and the presence of fixed complexes in spite of the medical treatment, are related with poor prognosis for the epileptic manifestations.


[Radicular syndrome caused by epidural tuberculous granuloma. Case report]. A case of tuberculous granuloma producing a radicular syndrome in a 58 year-old white male is reported. Excision of the granuloma and the administration of antituberculous therapy was followed by complete recovery.


Hypersomnia-sleep apnea due to micrognathia. Reversal by tracheoplasty. A 67-year-old woman with acquired micrognathia developed severe daytime hypersomnia, loud snoring, nocturnal enuresis, encopresis, and hypertension. A polysomnogram demonstrated 564 sleep apneas, primarily obstructive, recurrent hypoxia, a bradytachycardia, and absent stages III, IV, and REM sleep. Endoscopy during sleep revealed recurrent active closure of the upper pharynx associated with loud snoring. A tracheoplasty was done because of severity of symptoms and failure of conservative therapy. Dramatic improvement in sleepiness and hypertension occurred within 48 hours. On postoperative night 15 a repeated polysomnogram showed only 23 apneas, no hypoxia or bradytachycardia, and long periods of stage II, IV, and REM sleep. Patients with the hypersomnia-sleep apnea syndrome should be provided with a tracheal opening during sleep when severe daytime somnolence, cardiac arrhythmias, and hypertension are present.


Central retinal artery occlusion complicating Fabry's disease. A 16-year-old boy had a central retinal artery occlusion and was subsequently diagnosed as a hemizygote with Fabry's disease. The typical ocular manifestations in males with this inborn error of glycosphingolipid metabolism include whorl-like corneal epithelial infiltrates, retinal and conjunctival vessel tortuosity, and lenticular changes. The present case represents the first report of a retinal artery occlusion as an ocular complication of Fabry's disease.


Fibrous histiocytoma metastatic to the orbit. The term fibrous histiocytoma encompasses a heterogenous group of tumors believed to have a common origin in the histiocyte. These tumors demonstrate a wide histopathologic spectrum ranging from a predominantly cellular to a predominantly fibrous pattern. No clinical or histological criteria have been identified that can predict malignant behavior. Although generally characterized by local recurrence, a metastatic potential does exist. Fibrous histiocytoma of the thigh metastasized to the orbit.


Treatment of herpes simplex keratitis with levamisole. A model of chronic herpetic keratitis was developed by injecting rabbits subconjunctivally with a corticosteroid. A levamisole hydrochloride-treated group of these rabbits developed milder, more rapidly healing epithelial lesions than an untreated group. Although most of the untreated rabbits developed stromal disease, almost all of the lesions in the levamisole-treated animals were limited to the epithelium.


Transmission of a mucosal disease virus infection between sheep. The transmission of mucosal disease virus (MDV) from infected ewes and their lambs to susceptible sheep was investigated. MDV was recovered from the amniotic fluid of an infected pregnant ewe and from the blood, nasal swabs and urine of hairy lambs. MDV infection was transmitted either at lambing, from infective foetal fluids or lambs, or later as a result of contact with a surviving with a hairy lamb and either aborted or gave birth to an infected hairy lamb. Adult sheep and 12-months-old sheep were less readily infected than were newborn lambs. Pregnant ewes were infected by contact with a hairy lamb and either aborted or gave birth to an infected hairy lamb. A method to minimise spread of the infection in the field is suggested.


Epidemic adenovirus inclusions body hepatitis of the chicken in Australia. The occurrence of inclusion body hepatitis (IBH) on 11 properties in Australia is described. The 6 Victorian cases occurred between 1967 and 1973 and were identified as incidental findings accompanying other diseases or occurred as ill-defined, minor economic disease episodes. In well-managed flocks the extra mortality attributable to IBH was as low as 1.8% which represented a 50% increase in total mortality. The 1973 epidemic in New South Wales on 5 related farms resulted in flock mortality of 8.4% to 32.3% and an average mortality of 19.5% of 814,000 birds. For uninfected flocks of 307,000 birds the average mortality was 8.5%. Higher mortality in male flocks was statistically highly significant as was lowered efficiency of feed conversion. Growth rate in both sexes was depressed by about 0.15 kg in 70 days and heavy downgrading at processing occurred. Secondary infections were important. The cardinal features of IBH were fatal necrotising hepatitis with Cowdry type-A intranuclear inclusion body formation; secondary bacterial infections were also important causes of loss.


The pathogenesis of infectious avian encephalomyelitis. 1. The effect of the age of the chicken and the route of administration of the virus. The age of the chicken at the time of infection with infectious avian encephalomyelitis virus (IAEV) and the route of the administration of the virus had a marked effect on the development of the clinical disease. Chickens given the virus by intramuscular, intraperitoneal and oral routes exhibited a decreased susceptibility to development of clinical disease with increasing age. Irrespective of age, chickens were consistently susceptible to intracerebral inoculation of IAEV. Chickens infected with the virus at 1-, 7- and 14-days of age developed lower neutralising antibody titres to IAEV than chickens infected at 21 and 28 days. A relationship between the ability to produce specific neutralising antibody to IAEV and susceptibility to the development of clinical disease is discussed.


The pathogenesis of infectious avian encephalomyelitis. 2. The effect of immunosuppression on the disease. Normal chickens given infectious avian encephalomyelitis virus (IAEV) orally at 1, 7 or 14 days of age developed infectious avian encephalomyelitis (IAE), whereas those dosed with the virus at 21, 28 and 35 days of age did not. Chickens in all of these age groups that had been treated with cyclophosphamide or testosterone developed clinical IAE. Intraperitoneal inoculation of IAEV immunoglobulin at the time of dosing with the virus, or 48 hours later, protected normal and immunosuppressed chickens against the onset of clinical IAE. The titre of IAEV serum neutralising antibody was found to be lower in normal chickens given the virus at 1, 7 and 14 days of age in older chickens. These results suggest a significant role for the humoral immune system in the pathogenesis of IAE and in the development of immunity to the disease.


The pathogenesis of infectious avian encephalomyelitis. 3. The relationship between viraemia, invasion of the brain by the virus, and the development of specific serum neutralising antibody. An association was demonstrated between the development of clinical infectious avian encephalomyelitis (IAE), the persistence and titre of infectious avian encephalomyelitis virus (IAEV) in the brain of the chicken, the duration of detectable viraemia and the age of the chicken at the time of infection with the virus. The older the chicken at the time of infection the milder the disease, the lower the virus titre in the brain and the shorter the period of viraemia. IAEV serum neutralising antibody was produced earlier after infection in older chickens, and its detection was associated with decreasing virus titres in the brain and the cessation of detectable viraemia. Treatment of chickens with testosterone in ova, to inhibit the development of antibody synthesis, prevented the onset of age-associated resistance and testosterone treated birds were as susceptible to clinical IAE as baby chickens. The results suggested that the ability to produce IAEV serum neutralising antibody was an important component of age-associated resistance to IAE.


The pathogenesis of infectious avian encephalomyelitis. 4. The effect of maternal antibody on the development of the disease. Infectious avian encephalomyelitis virus (IAEV) maternal antibody was detected in the serum of chickens for up to 21 days following hatching. This antibody protected chickens against clinical IAE after intracerebral inoculation with van Roekel strain or oral administration of the NSW-1 strain of IAEV. Maternal antibody to IAEV also protected testosterone bursectomised chickens against the development of clinical disease. IAEV maternal antibody also influeced the pattern of virus excretion in faeces and serological responsiveness. This influence on antibody responses persisted beyond the time that IAEV maternal antibody could be detected. The importance of IAEV maternal antibody on the strategy of vaccination against IAE is discussed.


Lack of protection against oxygen toxicity by in vivo protective agents in the isolated toad bladder. Agents which protect against the development of oxygen toxicity in vivo were tested in an in vitro system, the isolated urinary bladder of the toad Bufo marinus. None of the agents protected against the inhibition of sodium transport across the bladder by hyperbaric oxygen exposure. Two protective agents, disulfiram and diethyldithiocarbamic acid, significantly increased the rate and extent of sodium transport inhibition by 5 ATA of oxygen, either when added in vitro or given in vivo. It was concluded that oxygen toxicity protective agents may have a nonspecific action in vivo.


Haemichrome formation from haemoglobin subunits by hydrogen peroxide. The effect of H2O2 on ferrous human haemoglobin subunits (alphash-, betash-, alphapmb- and betapmb-chains) was studied. These chains were easily transformed to haemichrome by the addition of H2O2 or H2O2-generating systems, including glucose oxidase (EC 1.1.3.4) AND XANTHINE OXIDASE (EC 1.2.3.2), and this was ascertained by e.p.r. measurements and by absorption spectra. The changes in these haemoglobin subunits were not inhibited by superoxide dismutase (EC 1.15.1.1), but were decreased by catalase (EC 1.11.1.6). The rate of oxidation of alphapmb-chains was higher than that of alphash-chains, and the rate of oxidation of betapmb-chains was higher than that of betash-chains. Haemichrome was demonstrated to be formed directly from these ferrous chains by the attack by H2O2, and this process did not involve formation of methaemoglobin. On the basis of these findings the kinetics of the reaction between the haemoglobin subunits and H2O2 was studied, and the pathological significance of H2O2 in disorders of erythrocytes such as thalassaemia was discussed.


Neutral metallo-proteinases of rabbit bone. Separation in latent forms of distinct enzymes that when activated degrade collagen, gelatin and proteoglycans. Rabbit bones in culture produce specific collagenase and neutral metallo-proteinase activity in latent forms that can be activated by either 4-aminophenylmercuric acetate or trypsin. Latent neutral metallo-proteinase activity was resolved by gel filtration into two enzymes, distinct from collagenase, that degrade gelatin and cartilage proteoglycans.


The temperature-dependence of the loss of latency of lysosomal enzymes. 1. When Triton-filled lysosomes from rat liver are incubated for up to 50min at 37 degrees C, pH7.4, in 0.25m-sucrose, no loss of latency of N-acetyl-beta-glucosaminidase or p-nitrophenyl phosphatase occurs unless the incubated lysosomes are cooled to approx. 15 degrees C. 2. It is suggested that a phase change takes place in the incubated lysosomal membranes on cooling; it starts at approx. 15 degrees C and probably is not complete at 0 degrees C. 3. Incubation of the lysosomes causes an increased potential for loss of latency of the lysosomal enzymes. This potential is not fully expressed at elevated temperature (e.g. 37 degrees C), but is expressed on cooling. 4. The increase at elevated temperature in potential for loss of latency exhibits biphasic kinetics, with an initial rapid phase followed by a slower phase, which is linear with respect to time. The extra loss of latency resulting from the rapid phase in proportional to the temperature of the incubation. 5. Arrhenius plots of the increase is potential for loss of latency during the slow phase for N-acetyl-beta-glucosaminidase and p-nitrophenyl phosphatase exhibit marked deviations from linearity beginning at approx. 15 degrees C. This suggests that the increase in potential for loss of latency is affected by a phase change that occurs around this temperature. 6. Activation energies for the increase in potential for loss of latency at and above 22 degrees C are 53.1+/-5.4kJ/mol (12.7+/-1.3kcal/mol) for N-acetyl-beta-glucosaminidase and 45.2+/-7.5kJ/mol (10.8+/-1.8kcal/mol) for p-nitrophenyl phosphatase. It is postulated that these energies reflect enzymic action, the products of which cause loss of latency to occur on cooling.


Complete loss of heparin-releasable triacylglycerol lipase activity after collagenase treatment of the rat liver. Lipolytic activity at pH 8.5-9.5 was lowered by approx. 80% in homogenates from livers perfused with collagenase or heparin. No heparin-releasable lipase activity was detected in hepatocytes isolated by collagenase treatment. It is concluded that crude collagenase completely inactivates the plasma-membrane-bound heparin-releasable lipase.


Effect of parathyrin on the transport properties of isolated renal brush-border vesicles. The transport properties of brush-border membrane vesicles isolated by a calcium-precipitation method from the renal cortex of normal and parathyrin (parathyroid hormone)-treated rats were studied by a rapid-filtration technique. Parathyrin elicited a dose-dependent decrease in the Na+-dependent phosphate uptake by the brush-border membrane vesicles, but the uptake of D-glucose, Na+ and mannitol was not affected. A maximum inhibition of 30% was observed after the application of 30 U.S.P. units intramuscularly 1 h before the animals were killed. Intravenous infusion of dibutyryl cyclic AMP (0.5-1.5 MG) also decreased the phosphate uptake by the brush-border vesicles. Both dibutyryl cyclic AMP and parathyrin were ineffective when added in vitro to brush-border membrane vesicles isolated from normal rats. These data suggest that parathyrin exerts its action on the phosphate reabsorption in the renal proximal tubule by affecting the Na+/phosphate co-transport system in the brush-border membrane. The effects of parathyrin on Na+ and glucose transport, however, seem to be due to alterations to the driving forces for transport and not to the brush-border transport systems.


Analysis of DNA structure by hydroxyapatite columns and ethidium bromide fluorescence techniques. A comparative study and effect on DNA binding. Seven duplex DNA preparations have been structurally analyzed by hydroxyapatite column chromatography and an ethidium bromide fluorescence technique. Significant contamination of one preparation with single-stranded DNA was detected by hydroxyapatite column chromatography. Five of the other six preparations were found to contain significant single-stranded regions by the ethidium bromide fluorescence technique. Synthetic poly dAT was found to be duplex in structure. The presence of single-stranded regions considerably influenced DNA binding results in a radioimmunoassay.


[Antiviral activity of plant components. 1st communication: Flavonoids (author's transl)]. Some drugs effective against influenza contain flavonoids. We therefore examined the antiviral effect of hesperidin, hesperidinmethylchalcon, trihydroxyethylrutin, catechol, quercitrin, rutin and aurantiin against vesicular stromatitis virus (VSV) action on mouse fibroblasts and that of hesperidin against influenza virus in HeLa cells system by means of dye uptake measurements (Finter) and by plaque reduction test, respectively. Preincubation of the cells with the flavonoids 6--8 h before virus addition was inevitable. Protection of cells against virus action persisted for about 24 h and it abruptly disappeared after an addition of hyaluronidase. Maximal inhibition of virus action was achieved with a concentration of 200 microgram/ml flavonoid.


[Studies of the possible importance of complement in increasing the hemagglutination-inhibiting activity of rabbit IgM against Sendai virus]. An investigation has been carried out in order to ascertain the effect of the presence of complement on the HI antibodies of the IgM class obtained in rabbit against Sendai virus. Contrary to the results obtained by some AA. with other viruses, in this case the presence of complement in the form of fresh guinea-pig serum has not modified the HI titre. It has however been demonstrated that in the guinea-pig serum an inhibitor is present, which is active towards Sendai virus and can be identified as a 19S alpha-globulin.


Technetium-99m stannous pyrophosphate scintigraphy in patients with calcification within the cardiac silhouette. Technetium-99m stannous pyrophosphate scintiscanning was performed in 22 patients with radiographically detected calcification within the cardiac silhouette. All but one of these scintigrams showed a localised area of increased activity similar to that ordinarily seen in acute myocardial infarction. Scintiscans in 3 patients after removal of the calcified aortic valve reverted to negative. It was concluded that this technique for acute infarct detection may yield false positive results in the presence of cardiac calcification.


Desensitization of the beta-adrenoceptor of lymphocytes from normal subjects and patients with phaeochromocytoma: studies in vivo. 1 Following the observation that lymphocyte beta-adrenoceptor responsiveness was not depressed in asthmatics treated only with non-adrenergic drugs we have explored the effects of prolonged exposure to beta-adrenoceptor agonists in normal subjects. 2 Treatment with oral salbutamol (12-16 mg/kg/day for 10 days), or with inhaled salbutamol (3000 microgram/day for 8-10 days) resulted in a significant reduction in lymphocyte beta-adrenoceptor responsiveness. 3 A 48 h infusion of isoxsuprine (10 mg/h) resulted in a marked depression of lymphocyte beta-adrenoceptor responsiveness (P less than 0.001). 4 Prolonged elevation of endogenous catecholamines caused by phaeochromocytoma was also associated with a marked depression of lymphocyte beta-adrenoceptor responsiveness (P less than 0.001). 5 There was no evidence that an increase in phosphodiesterase activity could explain the reduced cyclic AMP response. 6 It is concluded that diminished beta-adrenoceptor response occurs as a response to prolonged exposure to beta-adrenoceptor agonists. It is likely that the diminished response seen in asthmatic subjects can be explained on a similar basis and does not indicate an inherent cellular defect. 7 The possible clinical significance of such changes in asthmatics are discussed.


Desensitization of the beta-adrenoceptor of lymphocytes from normal subjects and asthmatic patients in vitro. 1 A lymphocyte culture method has been developed for studying in vitro the effect of prolonged exposure (24 h) to isoprenaline (10(-8) to 10(-6) mol/l) and prostaglandin E1 (PGE1; 2.8 x 10(-6) mol/l). 2 The cyclic AMP response to isoprenaline is reduced by prolonged exposure to isoprenaline. The degree of desensitization is in proportion to the concentration of isoprenaline in the culture medium. 3 Culture with isoprenaline does not reduce the cyclic AMP response to PGE1. 4 Culture for 24 h with PGE1 (2.8 x 10(-6) mol/l) reduces the cyclic AMP response to PGE1. It also significantly reduces the response to isoprenaline. 5 Lymphocytes from asthmatic patients show a similar degree of desensitization to isoprenaline (after 24 h culture with isoprenaline) to that seen in lymphocytes from normal subjects. 6 A modified assay for phosphodiesterase (PDE) activity was developed. PDE activity increased 24.5% (P less than 0.02) after culture with PGE1 but was not significantly affected by culture with isoprenaline. 7 It is concluded that desensitization caused by prolonged exposure to various stimulators of adenylate cyclase is an event dependent on several components, not all of which are yet defined.


Characterization of beta-adrenoceptor subtype mediating the metabolic actions of salbutamol. 1. The following four intravenous treatments were administered in a balanced, randomized, Latin square design to four healthy volunteers: (1) saline injection (10 min); salbutamol infusion (0.15 microgram kg-1 min-1 for 60 min) (sS), (2) propranolol injection (0.3 mg kg-1 for 10 min); salbutamol infusion (pS), (3) practolol injection (2 mg kg-1 for 10 min); salbutamol infusion (PS) and (4) saline injection; saline infusion (ss).  2. Heart rate was recorded and venous blood taken for estimation of insulin, glucose and potassium before and after each injection (0 and 12-15 min) and at various times during the infusion (30,45,60 and 75 min).  3. The mean, peak % heart rate changes from baseline, control were +65.8%, +23.0%, -10.9% and -12.1%, the mean, peak % glucose changes, +61.0%, +7.1%, +3.6% and +6.9%, and the mean, peak % insulin changes, +298%, +28%, -28% and -43% during the infusions for sS, PS, ss and pS respectively.  4. The mean serum potassium levels before the injections and at the completion of the infusions were 3.98 and 3.08 4.03 and 3.63, 4.07 and 4.15, and 4.03 and 4.13 meq/l for sS, PS, ss and pS respectively.  5. Propanolol completely abolished the cardiac and metabolic responses of salbutamol.  6. Practolol produced only partial cardiac beta-adrenoceptor blockade, but completely inhibited the metabolic actions of salbutamol.


Adenosine deaminase and adenosine kinase in rat hepatomas and kidney tumours. Adenosine deaminase and adenosine kinase have been measured in rat liver, 12 transplantable hepatomas, regenerating, foetal and neonatal liver, adult and neonatal rat kidney and 2 transplantable kidney tumours. Adenosine, deaminase activity, relative to the normal liver value, was elevated 2-4 fold in hepatomas of rapid growth rate, was in the normal range in more slowly growing hepatomas and in regernerating liver, and was low in foetal and neonatal liver. Adenosine kinase activity was decreased, relative to rat liver values, in all the hepatomas; activity of this enzyme gave a negative correlation with tumour growth rate. Kinetic properties of the two enzymes were examined in partially purified preparations. Adenosine deaminases from both liver and rapidly growing hepatoma 3924A were subject to weak product inhibition by inosine. Adenosine kinase from liver and hepatoma 3924A was inhibited by the reaction products ADP and AMP, and the enzyme was also subject to excess substrate inhibition by concentrations of ATP in excess of 1 mM. In rat hepatoma cell lines growing in culture, the toxicity of adenosine correlated inversely with the ratio of adenosine deaminase activity to adenosine kinase activity. Chromatographic measurements showed that hepatoma cells incorporated less extracellular adenosine into their adenine nucleotide pools than did isolated liver cells. These results indicate that increased adenosine deaminase activity and decreased adenosine kinase activity may confer a selective advantage upon the cancer cell.


Preparation and characterization of an antiserum to cultured human oat-cell carcinoma cells. Viable cultured oat-cell carcinoma cells were used to immunize a goat. The resulting antiserum contained high titres of anti-normal activity and antibodies to CEA. It was also shown, by indirect immunofluorescence, using fluorescein-conjugated rabbit anti-goat Ig, to localize at high titres on the surface membranes of human lung cancer cells of 4 different histological types. Booster immunizations produced a maximum secondary response one week after 2 weekly injections. The course of each immunization has been monitored for activity against normal human tissues, and the final sera have been absorbed with human spleen cells to remove anti-normal activity. Cross-reactivity with the lung-cancer-cell panel and antibodies to CEA persisted in high titre after absorption of anti-normal antibodies, and were present in the ammonium-sulphate-precipitated globulin fraction. The cells used for immunization did not produce detectable amounts of CEA in culture, and were not known to contain CEA prior to this experiment. Removal of anti-CEA antibodies by absorption with purified CEA has not reduced the cross-reactivity of the absorbed antiserum with the panel of human lung-cancer cells.


Solid tumour models for the assessment of different treatment modalities: VII: single vs fractionated doses of 5-fluorouracil on two solid tumours and their hosts. The effects of one large single dose of 5-fluorouracil (FU) have been compared to the same amount given in divided doses daily over a 3- or 5-day period. Comparison of the effects of single vs fractionated dosage was made on 2 types of experimental solid tumour with different growth, cell kinetic, histological and metastasizing properties. The tumour response was essentially the same for both the single and fractionated dose schedules.There were marked increases in animal mortality from drug toxicity following fractionated doses of FU compared to one large single dose. Mortality in animals with Tumour 3924A increased from 10% following one large single dose to 60% for animals given daily fractionated doses for 3 days, and 80% for animals given daily fractionated doses for 5 days. Total marrow reserve was measured by the total DNA content of tibial marrow. The nadir of 6 days for loss of total tibial marrow DNA following one large dose of FU was increased to 9 days for both fractionated schedules of FU. The 3-day delay in recovery of the marrow prevented recovery within the time frame necessary for animal survival. The inference from these experimental cancer-treatment studies is that daily fractions of chemotherapeutic agents such as FU result in increased morbidity and mortality, without benefit in the control of the solid tumour. The results question the advisability of the clinical practice of initially giving small daily "loading doses" of proliferation-dependent agents such as FU.These results emphasize the need for more precise information on the temporal relationship between the response and recovery of the host and the response and recovery of the solid tumour. They also emphasize the need for a better clinical understanding of the time sequence of solid-tumour recovery in relation to the time sequence of marrow recovery.


A fine structural study of the removal of the effectiveness of benzo-pyrone treatment of lymphoedema by the destruction of the macrophages by silica. Macroscopical, light microscopical and electronmicroscopical observations were made of the diaphragm, skin and brain of rats, some of which were treated with intraperitoneal silica for 8 days (after being given it i.v. for 2 days). The diaphragms showed a most remarkable increase in fibroblast activity and fibrosis beneath the peritoneal mesothelium (which was disintegrating). Deep to this there were many disintegrating macrophages, and much oedema and increased protein concentration. Ligation of the cervical lymphatics produced the usual changes of lymphoedema in the skin and brain. This was greatly reduced in the animals treated with a mixture of benzo-pyrones. However, in those animals also treated with silica, the benzo-pyrones had no effect on the amount of oedema or of protein. In all the animals except those treated with silica, lymphoedema was accompanied by considerable numbers of macrophages entering the affected tissues; in those treated with silica, these numbers were greatly reduced.


DNA unwinding protein from meiotic cells of Lilium. An ATP-dependent DNA unwinding protein is present at a high level of activity in meiotic cells of lilies. The protein also acts as a DNA-dependent ATPase, the single strand form being the preferred cofactor. It binds in the absence of ATP to single-strand DNA and to ends or nicks in duplex DNA. A 3'-OH terminus is required for binding at duplex ends; such binding is highly stable. Unwinding occurs in the presence of ATP, and it is limited to about 50 base pairs per end or 400-500 base pairs per nick. The ATP hydrolyzed during unwinding is distinguishable from ATP hydrolysis in the presence of single-strand DNA.


Chemical and enzymatic properties of riboflavin analogues. The chemical and enzymatic properties of 26 analogues of riboflavin are presented. These analogues include both endo- and exocyclically substituted isoalloxazines with redox potentials from -370 to -128 mV. Physical and chemical data such as the electronic absorption spectra, pKas, and redox potentials of the analogues are presented and are discussed with respect to preferred tautomeric and resonance forms. Like riboflavin, most of the analogues are shown to be catalytic oxidants of dihydro-5-deazaflavins. Analogue binding to egg white binding apoprotein has been quantitated and serves to determine the origins of binding site specificity for this protein. Nearly all of the analogues that possess D-ribityl groups are found to be processed to the FAD level by the flavokinase/FAD synthetase system of Brevibacterium ammoniagenes. Most extensively studied are the reactivities of the analogues with the NAD(P)H:flavin oxidoreductase of Beneckea harveyi. Many of the analogues are substrates in this enzymatic redox reaction, and a linear free energy-rate relation (log Vmax vs. E0' of the analogue) is seen that parallels similar relationships in the nonenzymatic oxidation of dihydro-5-deazaflavins. This suggests a common mechanism for the reactions of such diverse flavins as riboflavin, 5-deazariboflavin, and 1-deazariboflavin.


Spin-labeled ribonuclease A. Effects of chemical, enzymatic, and physical modifications on enzyme conformation. 3-SLHis-105-RNase A is an active derivative of ribonuclease A (RNase A) spin-labeled at the 3 position of the imidazole ring of histidine-105. The spin-labeled enzyme has been modified by urea denaturation, reduction, reduction-carboxymethylation, performic acid oxidation, and digestion with proteolytic enzymes in order to monitor changes in the geometry of the protein by changes in the electron paramagnetic resonance (EPR) spectrum of the nitroxide spin-label probe. The results of these experiments indicate that the spin-label attached to histidine-105 of RNase A is sensitive to modifications affecting the conformational integrity of the molecule and to the reconstituting effects of various active-center ligands.


Librational motion of an "immobilized" spin label: hemoglobin spin labeled by a maleimide derivative. The spin label Tempo-maleimide, when "immobilized" in hemoglobin, is shown to exhibit motional fluctuation whose amplitude and/or frequency depend on temperature and solution conditions. These motional fluctuations are observable by several electron spin resonance techniques. For desalted hemoglobin the fluctuations are detectable at approximately -15 degrees C using saturation transfer techniques and at approximately +25 degrees C using line-width measurements of normal absorption spectra. In ammonium sulfate precipitated hemoglobin, however, motional fluctuations are not detectable by either technique up to at least 40 degrees C. The most probable mechanism for spin-label motion appears to be either fluctuations in protein conformation which affect the label binding site or conformational transitions of the nitroxide ring itself. These motional fluctuations are shown to introduce a librational character to the overall label motion during hemoglobin rotational diffusion, with the librational motion significantly affecting the use of spin-label spectral shapes to calculate hemoglobin rotational correlation times.


Circular dichroism and circular polarization of luminescence of reduced nicotinamide adenine dinucleotide in solution and bound to dehydrogenases. The CD (circular dichroism) and CPL (circular polarization of luminescence) spectra of NADPH in aqueous solution were studied and found to be markedly different. The spectra were not affected by cleavage of the coenzyme molecule with phosphodiesterase. The differences are thus not due to the existence of extended and folded conformations of NADPH and it is concluded that they originate in excited state conformational changes of the nicotinamide--ribose fragment. Opposite signs of both the CD and CPL spectra were observed for NADH bound to horse liver alcohol dehydrogenase and to beef heart lactate dehydrogenase indicating structural differences between the nicotinamide binding sites. The binding of substrate analogues to enzyme--coenzyme complexes did not affect the CD spectra and hence no significant conformational changes are induced upon formation of the ternary complexes. No changes in the CPL spectrum of NADH bound to lactate dehydrogenase were observed upon adding oxalate to form the ternary complex. Marked differences were found between the CPL spectra of binary and ternary complexes with liver alcohol dehydrogenase, while the CD spectra of these complexes were identical. It is concluded that a conformational change of the excited NADH molecule occurs in the binary but not in the ternary complex involving LADH, thus indicating an increased  rigidity of the latter complex.


Mechanistic study of the addition of pyruvate to NAD+ catalyzed by lactate dehydrogenase. The binary complex of NAD+ and dogfish A4 lactate dehydrogenase reacts reversibly with pyruvate enol to produce an inactive, enzyme-adduct complex, in which the nicotinamide and pyruvate moieties are linked by means of a covalent bond. This process is examined in both the forward and reverse directions as a function of reactant and buffer concentrations at pH 7, under conditions where the enolization of pyruvate is at equilibrium, and the involvement of complexes with stoichiometry E.NAD, E.NAD.PyrE, and E.NAD.PyrK is defined. (The subscripts, E and K, indicate the enol and keto forms of pyruvate.) One pathway for formation of the adduct complex involves the prior formation of the E.NAD.PyrE complex from E.NAD and pyruvate enol; the alternative pathway involves formation of the same complex via the enolization of E.NAD.PyrK, a process that is catalyzed by an external (nonenzymic) base. The possible use of the adduct reaction as a model for the normal enzymic reaction is considered.


Isotope-tapping experiments with rabbit liver fructose bisphosphatase. Isotope-trapping experiments with mental-free rabbit liver fructose 1,6-bisphosphatase have shown that enzyme-bound D-fructose 1,6-bisphosphate completely dissociates prior to enzyme turnover initiated by Mn2+ as the catalytic metal. The exchange rate of the binary enzyme-D-fructose 1,6-bisphosphate complex with the substrate pool is, therefore, more rapid than its conversion to products, suggesting that structural Mn2+ is necessary for productive substarate binding. Rapid-quench isotope-trapping experiments confirm the requirement for structural Mn2+ ions for productive binding to occur. These experiments also show that an ordered formation of the enzyme-Mn2+ s-D-fructose 1,6-bisphosphate ternary complex which features metal-ion addition prior to substrate constitutes a catalytically competent pathway in the mechanism of fructose 1,6-bisphosphatase and that all four subunits are active in a single turnover event.


Spin-label and deuterium order parameter discrepancies in bilayers: one possible explanation. We have simulated electron spin resonance spectra of anisotropically immobilized spin labels of the type seen in lipid and soap-like bilayers using a rigorous formalism which explicitly includes the effects of spin-label motion. In most bilayer systems, spin-label experiments have shown lower order parameters then deuterium-label experiments. In the past this apparent decrease in the order parameters was thought to reflect the distortion of the bilayer by the doxyl ring of the spin probes. We wish to report that this type of discrepancy may be due to the neglect of important motional effects in the time-independent effective Hamiltonian formalisms used in previous interpretations of anisotropically immobilized spin label spectra. That the true order parameters may be the same can be shown by including slow motional corrections in the effective Hamiltonian formalism. The larger volume of the doxyl ring may change the apparent order parameter by increasing the importance of the slow motional effects, as opposed to causing a real decrease in the order parameter, as previously proposed.


Anomalies in the polarographic measurement of cytochrome oxidase activity. Reaction kinetics of the reduction of O2 by cytochrome oxidase follow essentially the same rate equation as that proposed for the oxidation of cytochrome c. However, the apparent second order rate constant varies with the oxidase concentration. The redox level of cytochrome c at the steady state was found to be essentially temperature-independent. Currently recognized pathways (or mechanisms) of electron transport from cytochrome c to O2 do not predict, and cannot account for the occurrence of these phenomena.


Characterization of the iron-sulfur protein of the mitochondrial outer membrane partially purified from beef kidney cortex. The iron-sulfur protein present in the mitochondrial outer membrane has been partially purified from beef kidney cortex mitochondria by means of selective solubilization followed by DEAE-cellulose chromatography. The EPR spectrum of the iron-sulfur protein with g-values at 2.01, 1.94 and 1.89 was well resolved up to 200 K which is unusual for an iron-sulfur protein. Analyses confirmed a center with two iron and two labile sulfur atoms in the protein. By measuring the effect of oxidation-reduction potential on the EPR signal amplitude, midpoint potentials at pH 7.2 were determined both for the purified iron-sulfur protein, +75 (+/- 5) mV, and in prepared mitochondrial outer membrane, +62 (+/- 6) mV. At pH 8.2 slightly lower values were indicated, +62 and 52 mV, respectively. The oxidation-reduction equilibrium involved a one electron transfer. A functional relationship to the rotenone-insensitive NADH-cytochrome c oxidoreductase in the mitochondrial outer membrane is suggested. Both this activity and the iron-sulfur center were sensitive to acidities slightly below pH 7 in contrast to the iron-sulfur centers of the inner membrane.


EPR properties of the reaction center of Rhodopseudomas gelatinosa in situ and in a detergent-solubilized form. The photochemical reaction centers from a variety of purple photosynthetic bacteria are composed of a trimer of protein subunits. However, the recently isolated reaction center from Rhodopseudomonas gelatinosa appears to have only two subunits. In this paper we examine the EPR characteristics of the primary photochemical reactants in this species, and compare them with those of other species. Despite of the differences in protein composition, no dramatic differences in EPR properties are seen in vivo, although some interesting effects are seen upon solubilization of the reaction center, which may be related to the unusual lability of the isolated preparation. Perhaps the most noteworthy phenomenon seen in Rps. gelatinosa is the apparent ability of electrons on the reduced intermediary electron carrier to tunnel at low temperatures to the oxidized c-type cytochrome, which has not been seen in other species studied to date.


Control of respiration in proteoliposomes containing cytochrome aa3. I. Stimulation by valinomycin and uncoupler. 1. Both valinomycin and p-trifluoromethoxy carbonyl cyanide phenylhydrazone (FCCP) are required for full release of respiration by cytochrome c oxidase-containing proteoliposomes (prepared by sonicating beef heart cytochrome aa3 in salt solution with 4 parts phosphatidylcholine, 4 parts phosphatidylethanolamine and 2 parts cardiolipin) in the presence of external ascorbate and cytochrome c. In the absence of valinomycin the response to FCCP is rather sluggish, as reported by Wrigglesworth et al. (1976) (Abstracts, 10th Int. Congr. Biochem., No. 06-6-230). 2. The Km for cytochrome c in 67 mM, pH 7.4, phosphate buffer with ascorbate as substrate, was 9 micrometer in both absence and presence of valinomycin and FCCP. Energization thus acts non-competitively towards cytochrome c oxidation. 3. The apparent Km for oxygen is greater in the energized than in the deenergized state; double reciprocal plots of respiration rate versus oxygen concentration are concave downward in the absence of uncouplers, as found with intact mitochondria. Energization thus acts "competitively" towards oxygen. 4. Despite the lack of a functional ATPase system, all the kinetic features of energization found in intact mitochondria can be mimicked in the reconstituted liposomes. This supports the chemiosmotic idea that electrical and perhaps H+ gradients modify the oxidase activity in reconstituted vesicles.


Purification and characterization of cytochrome O from Azotobacter vinelandii. The membrane-bound cytochrome O has been solubilized from the Azotobacter vinelandii electron transport particle and further purified by use of conventional chromatographic procedures. The spectral characteristics as well as the other properties noted for purified cytochrome O are reported herein.