[Electrophysiological effects of chloroacetyl ajmaline in Wolff-Parkinson-White syndrome]. The electrophysiological effects of chloro-acetyl-ajmaline in the Wolff-Parkinson-White syndrome have been studied in 7 patients after an intravenous dose of 1.5 mg/kg of the drug. Preexcitation was abolished in 3 cases, while 3 other subjects showed a slight increase in effective refractory period of the abnormal route of excitation (a mean of 13 ms). The possibility of bringing about a reciprocal rhythm was removed in one case out of two. During tachycardia, chloro-acetyl-ajmaline produced significant lengthening of the ventriculo-atrial conduction time (p less than 0.05). These results show the usefulness of chloro-acetyl-ajmaline in the control of the arrhythmias associated with the Wolff-Parkinson-White syndrome.


[Pulmonary arterial hypertension caused by neoplastic thrombosis of the pulmonary artery]. Neoplastic thrombosis of the pulmonary artery is a rare and little known cause of pulmonary arterial hypertension. The clinical picture is one of acute respiratory failure and progressive right ventricular failure caused by pre-capillary pulmonary hypertension. In the living patient there is no way of distinguishing this condition from that of subacute cor pulmonale due to embolism, especially as the primary tumour is not always found either because it is too small or because it has already regressed by the time it has metastasised. The diagnosis usually rests on histological examination of the lungs, and two pathological types can be distinguished: carcinomatous lymphangitis with secondary invasion and thrombosis of the pulmonary arterioles on the one hand, and the neoplastic arterial emboli of a chorio-epithelioma on the other.


[Coronary spasm on exercise. Demonstration of a case by coronary angiography]. We have seen a case with spasm of the right coronary artery coming on during exercise ergometry in the course of a coronary arteriogram. The patient had angina pectoris spontaneously and on exercise. Bicycle ergometry was repeated four times by the same method, and was positive in three; in two of these there was angina and ST elevation in II, III, aVF, and ST depression in I, aVL and V2 to V5. On one occasion the test was negative, the patient having taken a trinitrin tablet one hour before the test. Repeating the exercise test during coronary arteriography showed spasm of the right coronary artery and elevation of the segment ST in II, III and aVF; this disappeared after trinitrin treatment.


[A rare cause of continuous thoracic murmur. A fistula between the internal mammary artery and the pulmonary circulation]. The authors present a case of a continuous right lateral sternal murmur which led to the discovery of a systemic-to-pulmonary fistula at the origin of the internal mammary artery. The fact that the patient had a right pulmonary abscess with pyopneumothorax drained several years previously has led them to discuss the possible etiology of this fistula. The murmur disappeared after surgical treatment.


[Permanent atrial paralysis].

[Permanent atrial paralysis]. A case is described of permanent atrial paralysis in a 68 year old female with diabetes and cirrhosis of the liver. Absence of electrical atrial activity was expected on the standard electrocardiogram, and confirmed by intracavitary electrophysiological investigation which showed the ineffectiveness of stimulation of both the right atrium and the coronary sinus. Recording of the His potentials showed that the resultant rhythm was of supra-ventricular origin. The absence of mechanical atrial activity was confirmed radiographically, on the jugular venogram, on the apexogram, and on the tracings of right atrial and pulmonary capillary pressure. A search was made for muscular or neuromuscular dystrophy which has often been found in association with this arrhythmia, but none was found. There may be an etio-pathological relationship between carbohydrate metabolism and atrial paralysis, as certain authors have suggested.


[Surgical treatment by valve replacement and aorto-coronary bypass in mitral valve insufficiency caused by chronic dysfunction of the posterior papillary muscle]. Nine patients with cardiac failure which was refractory to medical treatment, and which was caused by chronic malfunction of the posterior papillary muscle, as a result of a myocardial infarction, were studied by cardiac catheterisation and coronary arteriography. The mean pulmonary capillary pressure was 31+/-16 mm of mercury with a nu wave at 51+/-27 mm of mercury. The end diastolic volume was increased (141+/-68 ml/m2) and the ejection fraction lowered (0.40+/-0.13). The left ventricle had overall hypokinesia in 5 patients and akinesia of the inferior wall, representing 21+/-24% of the end diastolic perimeter, in 3 others. All these patients had significant lesions of two or three of the main coronary trunks. At operation lengthening of the posterior papillary muscle and/or the cordae was found. All patients had a replacement mitral valve of the Starr-Edwards type, associated with an aorto-coronary bypass of the anterior descending artery. The operative mortality was zero. At a mean follow-up period of 21 months, there had been no late death, and all the patients were improved.


[Left ventricular aneurysms. Hemodynamic studies and surgical results]. The cardiac function of 37 cases of ventricular dyskinesia were studied by cineangiography. The amplitude and speed of contraction of the contracting parts was evaluated as well as the volume of the akinetic zone. A score of coronary artery obstruction was made for those patients who underwent coronary arteriography. All the patients had resection of their aneurysm. Four patients died in the immediate post-operative period. Six patients developed a temporary low output state requiring circulatory assistance. Twenty-seven patients were operated on successfully without posing any haemodynamic problem. Good immediate and late post-operative results were associated with an ejection fraction of the contracting part greated than 0,40, a dyskinetic area less than 40% and a coronary artery obstruction index of less than 6. Resection of the aneurysm, sometimes associated with aorto-coronary bypass grafting (5 cases), led to a marked improvement in heart failure and/or disappearance of ventricular arrhythmias.


[Effort electrocardiography and coronary arteriography following a 1st myocardial infarction. Critical study of the effort test]. Sixty-five patients, convalescent from a first myocardial infarction (anterior in 24 cases; inferior in 41 cases), underwent an effort electrocardiogram on a treadmill and coronary arteriography. In the anterior infarcts, coronary arteriography showed single vessel disease (anterior descending artery) in 54% of cases and double or triple vessel disease in the others. The effort test was positive in only 25% of patients with an anterior infarct. The presence of stenotic lesions of the circumflex artery and/or right coronary artery was unsuspected in 63% of patients. In the inferior infarcts, there was a significant stenosis of the anterior descending artery in 51% of the cases. The effort test was positive in 54% of patients and in 77% of those the anterior descending artery showed a significant stenosis. The appearance (or increase) of ST elevation greater than or equal to 1 mm in the leads facing the infarcted zone was an indication of more severe deterioration in left ventricular function as shown by a more marked reduction in ejection fraction and a more extensive akinetic region. The co-existence of ST elevation in the leads facing the infarcted zone and of ST depression greater than or equal to 1 mm in the reciprocal leads always indicated that another major vessel was involved, but this was only found in 25% of cases in this series.


[Obstructive cardiomyopathy and associated coronary atherosclerosis. Review of the literature and report of 2 personal cases]. The association of idiopathic hypertrophic subaortic stenosis (IHSS) with significant coronary atherosclerosis is little known, only 43 cases being available in the literature, 2 of which are personal ones. But the incidence of this association has certainly been underestimated. It is especially found from the sixth decade onwards, and at least 20% of patients with IHSS in and above the age group have stenosing lesions of the coronary artery. It is almost impossible to establish the presence of associated coronary abnormalities from the clinical features of from electrocardiogram. It does however seem worthwhile looking for this condition in IHSS when there is refractory chest pain, especially to beta-blockers, particularly if the patient is aged over 50 and has risk factors for ischaemic heart disease. It is also good to find IHSS associated with known coronary artery disease by using simple non-invasive techniques such as phonomechanocardiography and especially echo-cardiography; it is important not to miss the myocardial lesion and to treat concurrently if there is likely to be an indication for dealing with the coronary arteries surgically. The beta-blockers are the treatment of choice for both conditions, together with anticoagulents. If they fail, myectomy or myotomy together with aorto-coronary bypass graft should be considered.


[Intimal coronary fibrous dysplasia and Prinzmetal's angina]. It is currently considered as exceptional for there to be arterial pathology other than atheroma in Prinzmetal angina. The authors have found five cases of coronary fibrous dysplasia in the literature. They add a case, in a woman, of intimal fibrous dysplasia diffusely involving the three main coronary vessels. Coronary arteriography showed that spasm of the right coronary artery was the cause of attacks with the clinical and electrocardiographic features of Prinzmetal angina. The whole coronary tree was otherwise normal angiographically and also macroscopically at autopsy. The dysplasia was only discovered by histological study of serial sections of the three coronary vessels. Histological examination showed no evidence of an atheromatous lesion. The authors place these coronary artery lesions within the general classification of arterial fibrous dysplasias. It remains hypothetical whether there is a relationship between the occurrence of spasm and this very special histological feature of the coronary artery wall.


[Prognosis of adult angina pectoris by means of blood pressure and the electrocardiogram]. One hundred and fifty-three men with angina were studied to determine the risk of death over a 5-year period. Multivariate analysis using age, systolic and diastolic pressure, and six electrocardiographic variables (QRS axis, PR interval, the sum of S in V1 and R in V5, T-wave and ST segment anomalies and incomplete left bundle branch block) identified sub-groups having very different prognoses. An analysis using only six variables (omitting diastolic pressure, ST segment anomalies and PR interval) retained a good discriminatory value, and this same discriminant function calculated from half of the sample had good prognostic value in the other half. A prognostic index based on this function (designed to simplify the calculations in clinical use) identified a sub-group (24% of the total group) in which no mortality occurred, while another sub-group (16% of the total group) suffered a mortality of 67%. The logical use of the blood pressure and ECG leads to a more precise prognosis in angina and should help in determining the indications for myocardial revascularisation.


[Cor triatriatum associated with a ventricular septal defect in infancy. 2 cases treated surgically]. Although rare, the diagnosis of cor triatriatum in its classical form is usually made pre-operatively. The association with a large ventricular septal defect, as in the two cases reported here, masks the clinical and haemodynamic signs. In one case it was the echocardiogram and cineangiography which led to pre-operative diagnosis. The surgical correction of the two lesions was done successfully in one stage. In our second case, lack of recognition of the cor triatriatum led to the death of the patient in a low output state, following closure of the ventricular septal defect.


[Congenital arteriovenous fistula of the broad ligament. 1 case]. Following a case report of a congenital arterio-venous fistula of the broad ligament, the authors set out the different types of vascular malformation seen in the pelvic region and they attempt to define the place of treatment. The treatment of these lesions involves embolisation of the vascular pedicle and an attempt at surgical excision, alone or one after another.


[Recurrent ventricular tachycardia and parchment right ventricle in the adult. Anatomical and clinical report of 2 cases]. Two patients, aged 52 and 55 years, were admitted to hospital for attacks of recurrent ventricular tachycardia for which preventive treatment was difficult. The crises were of one type, with left-sided delay. The findings on clinical examination, electrocardiography, haemodynamic and arteriographic investigation and at post-mortem were those of the parchment right ventricle syndrome of the adult. Electrophysiological investigation in the 2 cases indicated that a re-entry mechanism underlay the attacks of tachycardia. Late activation of the pulmonary infundibulum was responsible for a post-excitation potential on the surface and intra-cavitary leads. Mapping carried out during ventricular tachycardia in one case demonstrated the pathway for re-entry at the level of the pulmonary infundibulum.


[Mitral valve insufficiency due to ballooning. A surgical series of 70 cases]. The authors report 70 cases of mitral regurgitation due to ballooning. For the surgeon this is the most frequent cause of degenerative mitral regurgitation in patients around 60-years-old. The aetiology still remains unknown. There are two opposing theories--the congenital and the acquired. The appearances at operation are characteristic--a localised or diffuse ballooning of one or both cusps with elongation of the chordae attached to the ballooned portion. Two histological features are noted--myxomatous degeneration of the cusp and the absence of any inflammatory process. Ballooning may be asymptomatic and bening for a long time before evolving progressively or suddenly into important mitral regurgitation. The pre-operative diagnosis is aided by left ventricular angiography and echocardiography. Surgical treatment, guided by the findings at operation, usually involves mitral valve replacement or, less frequently, mitral valve repair.


[Echocardiography in the diagnosis of mitral valve insufficiency due to valve mutilations or chordal ruptures. 41 cases with anatomical findings]. This study analyses the echocardiographic findings in 41 cases of severe mitral regurgitation due to chordal rupture (33 cases), elongation of chordae (4 cases) or valve trauma (4 cases). The operative findings are given. It was possible to make the diagnosis of chordal rupture in 60% of cases by recording one or more of the following signs:--For the anterior cusp: amplitude of motion equal to or greater than 38 mm; co-existence of chaotic diastolic fluttering and multiple systolic echoes; recording of several diastolic wave forms of the anterior cusp, out of phase and crossing each other;--For the posterior cusp: paradoxical movement of the cusp in systole and diastole; presence of an echo in the left atrium in systole. The group studied was compared with a group of 40 normal subjects and 48 cases of other types of mitral regurgitation which were severe and received surgery. The various signs had good specificity. The sensitivity of the different signs varied from 33 to 50% of cases. It was greater when the number of chordal ruptures was greater. Whichever cusp was affected, it was sometimes the site of high frequency and large amplitude systolic vibrations, which were found in a quarter of the patients. The specificity of this sign is discussed. The diagnosis of rupture of chordae is possible in a large proportion of cases and the causes of error are analysed.


Scanning electron microscope observation of the seminal vesicle in the Japanese monkey with special reference to intraluminal spermiophagy by macrophages. The seminal vesicle of the adult Japanese monkey was observed by scanning electron microscopy. A number of spermatozoa are found scattered on the luminal surface. Some of them are normal in appearance, while others disintegrated. The epithelial cells bulging slightly into the lumen are devoid of cilia, but provided with diffusely distributed microvilli. The fracture surface reveals that the epithelium of the seminal vesicle consists of columnar epithelial cells with secretory activity and occasional basal cells, in accordance with the finding obtained by transmission electron microscope in other mammals. The most striking observation of this study is the occurrence of a number of macrophages lying on the epithelium of the seminal vesicle. They also are seen to engulf the disintegrated spermatozoa in bulk. The intraluminal macrophages observed may serve to dispose of spermatozoa which have migrated into the seminal vesicle for some unknown reason.


Immunoperoxidase study in alpha-chain disease. Peroxidase-antiperoxidase technique was applied to search for the presence of alpha-chain protein in cells of the infiltrate from six cases of alpha-chain disease (one immunoblastic sarcoma and five plasmacytosis cases). Cases of poorly differentiated lymphocytic and Burkitt's type lymphomas, IgA myeloma, and tuberculous enteritis served as controls. Infiltrate cells from alpha-chain disease showed heavy and diffuse staining with anti-alpha-chain and not light-chain antisera. The myeloma reacted with both anti-alpha and anti-kappa antisera. Plasma cells from tuberculous enteritis showed variable staining with anti-heavy- and anti-light-chain antisera, while control lymphomas did not stain at all. We suggest that immunoenzyme histochemistry is a useful tool in demonstrating intracellular alpha-chain protein.


Perioperative problems in the diabetic.

Perioperative problems in the diabetic. Diabetes mellitus and surgery interact in two separate but important ways. Surgery has hormonal and metabolic consequences which tend to worsen the diabetic state. Diabetes has effects on blood vessels, and resistance to infection and wound healing, that tend to complicate surgery. In this review, the pathophysiological bases for these two interactions will be discussed, and practical methods of minimizing their consequences will be suggested.


External pancreatic fistula: a recent advance in management. The management of six consecutive recent cases of external pancreatic fistula is reported in which the use of total parenteral nutrition resulted in rapid closure of the fistula. The fistulae closed from 11 to 28 days after the commencement of total parenteral nutrition, with an average of 19 days. Surgical intervention to close the fistula was not required in any of the cases.


Coronary and myocardial metabolic effects of combined glyceryl trinitrate and propranolol administration. Observations in patients with and without coronary disease. Coronary haemodynamic and metabolic effects of propranolol and glyceryl trinitrate were studied in 12 patients with coronary artery disease and 5 without coronary heart disease, at rest and during tachycardia stress. Propranolol-associated reductions in indices of myocardial oxygen demand, left ventricle work, tension time, and left ventricle oxygen utilisation (LVVO2) were reversed when heart rate was controlled by atrial pacing. Adding glyceryl trinitrate at rest also restored heart rate but decreased the left ventricular work index and tension time index as coronary resistance declined paradoxically. Tachycardia-related increases in tension time index and LVVO2 were unchanged after propranolol, and ischaemia (angina, ST depression, and reduced lactate extraction) was not altered in most of the patients. During tachycardia, the addition of glyceryl trinitrate decreased the tension time index and LVVO2; angina recurred in only 4 patients, and ST depression and lactate extraction improved. Similar haemodynamic changes occurred in the patients with normal coronary arteries. In contrast with propranolol administered alone, propranolol plus glyceryl trinitrate enhances tachycardia tolerance and prevents tachycardia-induced manifestations of ischaemia. This action is attributed to glyceryl trinitrate-associated improvement in the adequacy of myocardial perfusion.


Semi-automated detection of emesis in the rhesus monkey. The use of a video recorder and the inclusion of a marker dye in material fed to rhesus monkeys has eliminated the need for the constant presence of an observer to detect an emetic response.


Inhibition of the growth of murine tumour cells in vitro by serum from non-immune syngeneic and allogeneic mice. Sera from DBA/2 and Quackenbush mice (which are non-immune for mastocytoma and Sarcoma 180 respectively) contain a heat-labile (56 degrees for 30 min) component(s) that inhibits the in vitro growth of DBA Mastocytoma P-815 X-2 and Sarcoma 180. Adsorption of the sera with tumour cells at 4 degrees did not eliminate the factor(s), suggesting that it is not an antibody. In liquid suspension cultures inhibitory activity was observed at concentrations of mouse serum of 10--20% and in semisolid agar clonogenic cell assays at concentrations as low as 1%. The influences of the inhibitor(s) for both tumours and in both culture systems were parallel. However, there was a quantitative difference in susceptibility to other environmental factors (FCS concentration, bicarbonate concentration, and O2 tension) between the two tumours. These results parallel the in vivo findings where intravenously injected mastocytoma cells produced more tumours than did Sarcoma 180.


Neurofibromatosis and pregnancy.

Neurofibromatosis and pregnancy. Ten patients with neurofibromatosis were studied in 27 pregnancies. Two of the patients had evidence of pre-eclampsia in their first pregnancy but not in subsequent ones. In all other pregnancies, no adverse features were noted. It would seem that neurofibromatosis is not specifically associated with any obstetric complications. The need for further reporting of cases is stressed.


Pathogenesis and classification of massive periretinal proliferation. Massive periretinal proliferation (MPP), a serious complication of retinal detachment, is caused by proliferation and fibrous metaplasia of cells mostly deriving from retinal pigment epithelium and retinal glial cells. Contracting fibrous membranes in the vitreous, and on and also under the retina, cause the intraocular changes of MPP. Early signs such as increased 'tobacco dust', pigmented and unpigmented clumps in the vitreous, and subtle preretinal and even retroretinal membranes are usually overlooked. The late signs such as starfolds, irregular retinal folds, circumferential folds, and funnel-shaped detachments are well known. The pathogenesis of the clinically visible signs is described, and a 4-stage classification of the disease is given.


Kinetic study on the reaction mechanism of pantothenase: existence of an acyl-enzyme intermediate and role of general acid catalysis. A kinetic study was performed on the reaction mechanism of pantothenase (EC 3.5.1.22) catalyzed hydrolysis of the pantothenic acid. A nonlinear progress curve is derived if the reaction occurs at low buffer concentrations. The nonlinearity is due to partial reversibility of the reaction; an acylenzyme (pantoyl-enzyme) is formed during the reaction, and beta-alanine, the other end product, is able to react with the acyl-enzyme and return back to pantothenate. The dependence of the beta-alanine return reaction on buffer concentration and on pH suggests a general acid catalysis during the reaction. A reaction mechanism is suggested, in which the -NH3+ form of beta-alanine participates in the return reaction, and the deacylation of the acyl-enzyme is acid catalyzed.


Partial purification and evidence for multiple molecular forms of the scrapie agent. A procedure for the partial purification of the scrapie agent from mouse spleen was developed based on its sedimentation profile. Differential centrifugation and detergent treatment with sodium deoxycholate yielded a fraction designated "P5" which was enriched for scrapie infectivity approximately 20-fold with respect to cellular protein. The P5 fraction was devoid of cellular membranes but heavily contaminated with ribosomes as judged by electron microscopy. On centrifugation of the fraction P5 to near equilibrium in a sucrose gradient scrapie infectivity was distributed over a range of densities from 1.08 to 1.30 g/cm3. Parallel rate-zonal analysis showed that the infectivity was distributed over a range of particle sizes with s20.w values from approximately 40 S to greater than 500 S. Incubation of P5 at 37 or 80 degrees C, under conditions that disrupt ribosomes, dramatically altered the rate-zonal gradient profile of the agent. Under these conditions, the agent sedimented as particles with s20.w greater than 500 S. The apparent heterogeneity of the scrapie agent with respect to both size and density and its ability to shift from one form to another suggest that the agent may contain hydrophobic domains on its surface.


Properties of the nitrogenase system from a photosynthetic bacterium, Rhodospirillum rubrum. Soluble nitrogenase from Rhodospirillum rubrum has been isolated and separated into its two components, the MoFe protein and the Fe protein. The MoFe protein has been purified to near homogeneity and has a molecular weight or 215 000. It contains two Mo, 25--30 Fe and 19--22 acid-labile sulphide and consists of four subunits, Mw 56 000. The Fe protein has a molecular weight 65 000. It contains approximately four Fe and four acid-labile sulphide and consists of two subunits, Mw 31 500. The highest specific activities for the purified components are 920 and 1260 nmol ethylene produced per min per mg protein, respectively. The purified components require the membrane component for activity (Nordlund, S., Eriksson, U. and Baltscheffsky, H. (1977) Biochim. Biophys. Acta 462, 187--195). Titration of the MoFe protein with the Fe protein shows saturation and excess MoFe protein over Fe protein is inhibitory. Addition of Fe2+ or Mn2+ to the reaction mixture increases the activity apparently through interaction with the membrane component.


Fluorescence from sensitizing phycobilin chromophores in the blue-green alga Anacystis nidulans. Regeneration of the pigment system of Anacystis nidulans was studied following nitrate starvation. Three new, distinct fluorescence bands, at 596, 615 and 636 nm attributed to sensitizing phycobilin chromophores were detected. They each possess a separate excitation band at 425, 395 and 410 nm, respectively.


Calcium and photosynthetic oxygen evolution in cyanobacteria. Calcium activation of oxygen evolution from French-press preparations of Phormidium luridum is largely reversible upon removal of added Ca(2+). Activation occurs via a first-order binding with a dissociation constant of 2.8 mM. An 8-fold increase in oxygen evolution rate observed upon Ca(2+) addition is accounted for by a 4-fold increase in the number of active photosynthetic units, and a doubling of turnover rate. While both Ca(2+) and Mg(2+) stimulate turnover, unit activation is Ca(2+) specific. Under optimal conditions, 30% of the units functioning in the intact cell can be recovered in the Ca(2+) -activated preparation. The Ca(2+) requirement of P. luridum preparations is not relieved by proton-carrying uncouplers, or by rate-saturating concentrations of the Hill acceptor, ferricyanide. Taken together with the reported stimulation by Ca(2+) of oxygen evolution in the presence of DCMU (Piccioni, R.G. and Mauzerall, D.C. (1976) Biochim. Biophys. Acta 423, 605--609) these observations strongly suggest a site of Ca(2+) action within Photosystem II. The pronounced specificity of the Ca(2+) requirement appears in preparations of other cyanobacteria (Anabaena flos-aquae and Anacystis nidulans) but not in the eucaryote Chlorella vulgaris. While milder cell-disruption methods bring about some Ca(2+) dependence in P. luridum, French-press treatment is required for maximal expression of Ca(2+) -specific effects. French-press breakage causes a release of endogenous Ca(2+) from cells, supporting the view that added Ca(2+) restores oxygen evolution by satisfying a physiological requirement for the cation.


A high-potential redox component located within cyanobacterial photosystem II. The calcium-dependent oxygen evolution activity of preparations of Phormidium luridum shows a marked selectivity in favor of ferricyanide over benzoquinone as Hill oxidant. In addition, the rate of oxygen evolution increases with increasing solution redox potential over the range +350 to +550 mV vs. the standard hydrogen electrode. These properties pertain to both 3-(3,4-dichlorophenyl)-1,1-dimethylurea-sensitive and -insensitive fractions of the total oxygen evolution activity. Neither changes in solution potential nor use of oxidants other than ferricyanide obviate the need for added Ca(2+). To explain these observations, two models are proposed, each of which invokes the existence of a redox component located within Photosystem II and having a midpoint potential greater than +450 mV. In one model, the postulated species is a donor which competes with water for oxidizing equivalents generated by System II. In the other model, the 450 mV species is a high-potential primary acceptor of System II electrons.


Excitation energy transfer between pigment system II units in blue-green algae. Efficiency in excitation energy transfer from closed to open reaction center II in blue-green and red algae was estimated by the method developed by Joliot and Joliot (C.R. Acad. Sci. (1964) 258, 4622--4625) after slight modification; the number of open reaction centers II was counted from the mean O2 yield of repetitive short flashes. The efficiency in energy transfer in Chlorella pyrenoidosa was the same in our measurement as that reported by Joliot and Joliot (0.55 +/- 0.02). However, the values obtained with four blue-green algae and one red alga were very small, in a range of 0.00--0.07. The low efficiency was always obtained independently of the size of the apparent photosynthetic unit which was varied by growth conditions. Results indicated that pigment system II forms a unit in which only one reaction center II is operative.


Polymorphic phase behaviour of cardiolipin as detected by 31P NMR and freeze-fracture techniques. Effects of calcium, dibucaine and chlorpromazine. 1. The influence of Ca2+ on the polymorphic phase behaviour of cardiolipin has been investigated employing 31P NMR and freeze-fracture techniques. The close correlation between the results obtained here and previous X-ray studies (Rand, R.P. and Sengupta, S. (1972) Biochim. Biophys. Acta 255, 484--492) confirms 31P NMR as a useful analytical procedure for investigating the polymorphic phase behaviour of hydrated phospholipids. 2. Ca2+ induces formation of the hexagonal (H11) phase via an intermediary phase which is observed at Ca2+/cardiolipin ratios of less than 1 (mol/mol). This intermediary appears to consist of "inverted' structure which lies adjacent to regions of bilayer structure. 3. The local anaesthetics dibucaine and chlorpromazine produce similar phase changes for cardiolipin as does Ca2+. It is suggested that the anaesthetics interact with the membrane in their charged form and induce their effects by charge neutralization.


Interaction of Pseudomonas cytochrome cd1 with the cytoplasmic membrane. Labelling with ferritin-conjugated antibody shows that Pseudomonas cytochrome cd1 is associated with the inner surface of the cytoplasmic membrane. Cytochrome cd1 is however, enriched to the soluble fraction obtained after destruction of Pseudomonas spheroplasts. Comparison of the respiratory nitrite reductase activities, due to this cytochrome, between different cellular fractions and the purified enzyme shows that while the kinetic pattern and the temperature dependence of the activity remain almost the same the molecular activity is enhanced when the enzyme is released from cells. A new assay of respiratory nitrite reductase was developed in this study. The method is based on determination of the stoichiometrical proton consumption accompanying nitrite reduction.


The nature and location of Acholeplasma laidlawii membrane proteins investigated by two-dimensional gel electrophoresis. The high resolution, two-dimensional electrophoresis system for the separation of proteins described by O'Farrell, (O'Farrell, P.H. (1975) J. Biol. Chem. 250, 4007--4021) has been modified for the separation of Acholeplasma laidlawii proteins. Reproducible protein patterns have been obtained from A. laidlawii cell, membrane and soluble protein preparations. The isoelectric focusing of membrane proteins was greatly improved by removing the bulk of the membrane lipid before solubilizing the protein. A. laidlawii peripheral membrane proteins were removed from the membrane by low ionic strength washing and by treatment with EDTA. The effect of an exhaustive EDTA treatment and a rapid, warm EDTA treatment were compared. By comparing the protein patterns obtained in these ways it was possible to distinguish two separate groups of peripheral membrane proteins and one integral membrane protein group. The peripheral membrane proteins which were removed from the membrane at low ionic strength (group I) were also insoluble in Triton X-100, whereas additional peripheral membrane proteins extractable by subsequent EDTA treatment (group II) were soluble in Triton X-100. Exterior-facing membrane proteins were distinguished from the interior-facing ones by lactoperoxidase-catalyzed iodination of intact cells and membranes. Group I peripheral membrane proteins faced the cell interior whereas group II proteins faced the cell exterior. We counted approximately 320 individual whole cell proteins. Of these, about 140 were membrane associated and a maximum of 40 proteins were iodinated after iodinating intact cells. A. laidlawii was also grown in the presence of NaH232PO4 and whole cell proteins were separated by two-dimensional gel electrophoresis. One membrane protein and two soluble proteins were labelled.


Concanavalin A promotes the uptake of lysosomal hydrolases by human fibroblasts. Human placental hexosaminidase B and beta-galactosidase are taken up very poorly by human fibroblasts in culture. However, if fibroblasts manifesting genetically determined deficiencies of these lysosomal hydrolases are first treated with concanavalin A, then enzyme uptake is markedly increased. Enzyme activity which becomes associated with concanavalin A-treated fibroblasts maintained at 4 degrees C can be greatly removed by treatment with haptene sugar, while enzyme activity which becomes associated with cells maintained at 37 degrees C is refractory to haptens treatment. These results are interpreted as an initial binding of enzyme to concanvalin A molecules located at the cell surface, followed by an active cellular process leading to internalization of the lectin-enzyme complexes.


Accessibility of phospholipids in the chromaffin granule membrane. 1. The accessibility of phospholipids in the membrane of the adrenomedullary storage vesicles (chromaffin granules) has been studied. 2. The reaction of 2,4,6-trinitrobenzenesulphonic acid with both intact granules and their ghosts, results in the labelling of 70% of the phosphatidylethanolamine. 3. The action of phospholipase A2 (from bee venom), phospholipase C (from Bacillus cereus) and sphingomyelinase C (from Staphylococcus aureus) on granules and their ghosts was followed as a function of time. No significant difference was observed between the intact granules and their ghosts. 4. In the intact granules the various treatments led to varying amounts of lysis although again no evidence was obtained that such lysis in any way increased the amount of accessible phospholipid. 5. Highly purified granule preparations were also compared with the so-called "large granule" fraction and no significant differences were detected. 6. Approx. 67% of phosphatidylethanolamine + phosphatidic acid, 50% of phosphatidylserine + phosphatidylinositol, 65% of phosphatidylcholine and 20% of sphingomyelin is accessible to enzymatic degradation. In total, approx. 50% of all the phospholipids reacted. 7. It is also shown that, unlike in enzymatic treatment, all the phosphatidylcholine can be exchanged in the presence of a phospholipid exchange protein (prepared from beef liver). 8. It is concluded that transmembrane movement of phosphatidylcholine is slow in isolated membranes of chromaffin granules. The presence of the exchange protein, however, in conjunction with membrane proteins and specific phospholipid arrangements may catalyse this transmembrane movement.


A cloned Drosophila DNA fragment which codes for a 4 S RNA species. A collection of random Drosophila melanogaster DNA fragments cloned individually in Escherichia coli was screened for the presence of sequences complementary to the 4 S, 5 S and 5.8 S RNA species produced in the D. melanogaster Kc tissue culture line. Four D. melanogaster DNA fragments were found which possessed sequences complementary to the 4 S RNA species but not complementary to the 5 S or 5.8 S RNA. One such cloned fragment (6.81 kilobase in length) was characterized further. It hybridizes in situ to region 22A-C of the left arm of chromosome 2 and does not contain repetitive sequences detectable by renaturation (cot) analysis. This same region was reported earlier by Steffensen and Wimber (Genetics (1971) 69, 163--178) to hybridize in situ to bulk tRNA extracted from D. melanogaster.


The use of standard and relaxed hybridization conditions to detect two classes of sequences related to type-D retroviruses in the DNAs of primates. Using standard hybridization conditions (68 degrees C and 0.4 M sodium phosphate) and assaying for RNAase-resistant RNA . DNA duplexes in the presence of 2 X SSC (1 X SSC is 0.15 M NaCl and 0.015 M sodium citrate), sequences representing approx. 20% of the Mason-Pfizer virus (MPV) genome have previously been shown to be endogenous in DNAs of all Old World monkeys examined (Drohan, W., Colcher, D., Schochetman, G. and Schlom, J. (1977) J. Virol. 23, 36--43). We now report that titration of both the temperature at which hybridizations are carried out and the Na+ concentration at which 125I-labeled MPV RNA . DNA hybrids are scored, reveals a second class of sequences related to the MPV genome in the DNAs of primates. These MPV-related sequences, which are similar to an additional 40% of the MPV genome, are detected when the temperature of hybridization is reduced to 54 degrees C and when the resulting 125I-labeled RNA . DNA duplexes are scored for RNAase resistance in 8 X SSC. These sequences are found in the DNAs of all Old World monkeys examined, and the Tm values of the hybrid duplexes are approx. 6--7 degrees C lower than those of the hybrid duplexes formed using standard conditions. These studies further demonstrate the wide distribution of type-D retrovirus sequences in primates.


Induction of sporulation by inhibitory purines and related compounds. Sporulation of Bacillus subtilis can be induced, in the presence of excess ammonia, glucose and phosphate, by many purine derivatives under conditions of partial growth inhibition. Some of the compounds are known inhibitors of purine nucleotide synthesis. For most compounds the effect is counteracted by adenine and guanine. Partial growth inhibition by amethopterin (methotrexate) causes sporulation in the absence of purines but not in their presence. Unable to induce sporulation at any concentration are inhibitors of DNA, RNA, and protein synthesis as well as base or amino acid analogs that are incorporated into these polymers.


The organization of ribosomal RNA genes in the mitochondrial DNA of Tetrahymena pyriformis strain ST. 1. We have constructed a physical map of the mtDNA of Tetrahymena pyriformis strain ST using the restriction endonucleases EcoRI, PstI, SacI, HindIII and HhaI. 2. Hybridization of mitochondrial 21 S and 14 S ribosomal RNA to restriction fragments of strain ST mtDNA shows that this DNA contains two 21-S and only one 14-S ribosomal RNA genes. By S1 nuclease treatment of briefly renatured single-stranded DNA the terminal duplication-inversion previously detected in this DNA (Arnberg et al. (1975) Biochim. Biophys. Acta 383, 359--369) has been isolated and shown to contain both 21-S ribosomal RNA genes. 14 S ribosomal RNA hybridizes to a region in the central part of the DNA, about 8000 nucleotides or 20% of the total DNA length apart from the nearest 21 S ribosomal RNA gene. 3. We have confirmed this position of the three ribosomal RNA genes by electron microscopical analysis of DNA . RNA hybrid molecules and R-loop molecules. 4. Hybridization of 21 S ribosomal RNA with duplex mtDNA digested either with phage lambda-induced exonuclease or exonuclease III of Escherichia coli, shows that the 21-S ribosomal RNA genes are located on the 5'-ends of each DNA strand. Electron microscopy of denaturated mtDNA hybridized with a mixture of 14-S and 21-S ribosomal RNAs show that the 14 S ribosomal RNA gene has the same polarity as the nearest 21 S ribosomal RNA gene. 5. Tetrahymena mtDNA is (after Saccharomyces mtDNA) the second mtDNA in which the two ribosomal RNA cistrons are far apart and the first mtDNA in which one of the ribosomal RNA cistrons is duplicated.


Conservation of the sequence and position of the ribosomal RNA genes in Tetrahymena pyriformis mitochondrial DNA. 1. We have done cross-hybridizations between the mitochondrial ribosomal RNAs and DNAs from strains ST and PP of Tetrahymena pyriformis. DNA . ribosomal RNA hybrid formation can be completely prevented by an excess of the heterologous ribosomal RNA and the heterologous hybrids melt 6 degrees C below the homologous hybrids. This shows that the ribosomal RNA cistrons can account for the 5% cross-hybridization previously observed between the mtDNAs of strains PP and ST (Goldbach et al. (1977) Biochim. Biophys. Acta 477, 37--50). 2. By electron microscopy of DNA . ribosomal RNA hybrids we have determined the position of the ribosomal RNA cistrons on the mtDNA of strain GL, a mtDNA which we have shown to contain a sub-terminal 1 micron duplication-inversion and a terminal palindrome at one end which varies in length from 0 to 5 micron and which includes the 1 micron duplication-inversion (Arnberg et al. (1977) Biochim. Biophys. Acta 477, 51--69). The 21 S ribosomal RNA cistron overlaps the 1 micron duplication-inversion and as a result two or three cistrons are present, depending on the size of the terminal palindrome. Only one 14 S ribosomal RNA cistron is found, located about 10 000 base pairs away from the nearest 21 S cistron is found, located about 10 000 base pairs away from the nearest 21 S cistron and with the same polarity as this cistron. 3. We conclude from these results and those in the preceding paper that the sequence of the ribosomal RNAs and the position of the ribosomal RNA genes in the mtDNA is strongly conserved in Tetrahymena. Possible reasons for the duplication of 21-S ribosomal RNA genes and the terminal heterogeneity of Tetrahymena mtDNA are discussed.


Primary structure of bovine liver tRNATrp. Purified tRNATrp from bovine liver, accepting 1700 pmol tryptophan per A260nm unit, was completely digested with pancreatic ribonuclease and T1 ribonuclease. The sequences of the resulting oligonucleotides were determined and the primary structure of the tRNA was deduced. These analyses showed numerous incomplete post-transcriptional modifications, and several positions heterogenously occupied by two different nucleotides, which lead us to think that in bovine liver there exist a mixture of several tRNATrp.


Phenobarbitone-induced stimulation of protein synthesis in liver of diabetic rat. The effect of phenobarbitone on liver weight, on the rate of protein synthesis and on the sedimentation profiles of polyribosomes from livers was studied in diabetic rats. The rate of protein synthesis by isolated postmitochondrial supernatants from diabetic rats is lower than that from normal animals. The analysis of polyribosome profiles and the effect of Sephadex chromatography on protein synthesis demonstrated that the reduction was dependent in part on polyribosomal disaggregation and in part on the presence in the cytosol of low molecular weight inhibitor(s). Phenobarbitone administration had the same effect in either diabetic or normal rats in that it increased, (a) the degree of polyribosomal aggregation, (b) the rate of protein synthesis by the isolated postmitochondrial supernatants, (c) liver weight and (d) the activity of the inducible enzyme, NADPH-cytochrome c reductase. Both polyribosomal and soluble factors appear to be involved in the phenobarbitone effect. As the diabetic rats do not secret insulin the results suggest that insulin is not involved in the control of protein synthesis by phenobarbitone. It is suggested that the intracellular redox state has a major influence on the rate of protein synthesis.


Studies on extracellular proteases of Streptococcus sanguis. Purification and characterization of a human IgA1 specific protease. Extracellular caseinolytic activity was found in the culture fluid of Streptococcus sanguis ATCC 10556 grown in a dialyzed culture medium. This activity was due to multiple proteases that differed in their elution from hydroxyapatite, sensitivity to enzyme inhibitors, specificity and optimum pH. IgA protease, which splits human immunoglobulin A1 into intact Fc and Fab could be effectively separated from these relatively non-specific proteases and purified to apparent homogeneity in 20% yield by a five-step procedure. Although the bulk of the dextran sucrase activity was separated from the IgA protease, a small amount of sucrase activity remained with the final IgA protease preparation. In polyacrylamide gel electrophoresis at pH 9.5 both activities were located in the single protein band detected in this preparation. A quantitative method for the assay of IgA protease was developed, based on radial immunodiffusion to quantitate the Fab produced. This was used to follow the specific activity and yield during purification, and to characterize some of the catalytic properties of the enzyme. At an enzyme/substrate ratio of 1: 400 (w/w) the protease could effect 50% proteolysis of IgA in overnight incubation at 37 degrees C. The optimum activity was at pH 8.0, and 50% inhibition was achieved at 4 . 10(-4) M o-phenanthroline or 8 . 10(-4) M ethylene diamine tetraacetate. Concentrations of diisopropyl phosphofluoridate, phenylmethyl-sulfonyl fluoride, iodoacetate and p-chloromercuribenzoate up to 10(-2) M were without effect on the IgA protease activity. Full reactivation of the chelator inhibited enzyme could be achieved by the addition of Mg2+, Mn2+ or Ca2+.


Subunit interactions and immobilised dimers of human liver arginase. Incubation of soluble human liver arginase (L-arginine amidinohydrolase, EC 3.5.3.1) with p-hydroxymercuribenzoate resulted in the dissociation of the enzyme into active dimers. Addition of 2-mercaptoethanol resulted in the regeneration of the tetrameric enzyme. When arginase, bound covalently to nylon, was incubated with p-hydroxymercuribenzoate, matrix-bound dimers were obtained. Incubation of these species with 2-mercaptoethanol resulted in stable, unmodified dimers. Based on this dissociation of arginase, a model with D2-symmetry is suggested for this enzyme. The specific activity, the Km value for arginine, pH optimum and the inhibition constants for ornithine and lysine were determined for monomeric, dimeric and tetrameric forms. It is concluded that the behaviour of the active sites of the monomers is not substantially altered by the interaction of these species in the oligomeric molecule.


Studies on the reaction intermediate of protocatechuate 3,4-dioxygenase. Formation of enzyme-product complex. The nature of the oxygenated intermediate observed (Fujisawa, H., Hiromi, K., Uyeda, M., Okuno, S., Nozaki, M. and Hayaishi, O. (1972) J. Biol. Chem. 247, 4422--4428) during the reaction of protocatechuate 3,4-dioxygenase (protocatechuate:oxygen 3,4-oxidoreductase (decyclizing), EC 1.13.11.3) was investigated. 3,4-Dihydroxyphenylpropionic acid and 3,4-dihydroxyphenylacetic acid were used as substrates of the enzyme to slow down the rate of the reaction. The enzyme reactions were performed under conditions where the concentration of the organic substrate was lower than those of the enzyme and oxygen in the reaction mixture. The reactions were stopped before completion by the addition of hydrochloric acid or guanidine hydrochloride and then the organic compounds were extracted from the reaction mixture to be analyzed. The qualitative analyses by thin-layer chromatography revealed that there was no species other than the organic substrate and the enzymatic reaction end-product during reaction. The quantitative spectrophotometric analyses revealed that the organic substrate which had participated in the formation of the oxygenated intermediate existed as a species indistinguishable from the reaction end-product, indicating that the oxygenated intermediate was not a simple complex of oxygen, substrate and the enzyme, i.e., a ternary complex, but a species rather close to a binary complex of product and the enzyme.


Identification of pseudoisoenzymic subforms of muscle carbonic anhydrase. Rabbit muscle carbonic anhydrase III, a recently discovered third isoenzyme (possibly muscle specific) of carbonic anhydrase (carbonate hydro-lyase, EC 4.2.1.1) (Register, A.M., Koester, M.K. and Noltmann, E.A. (1978) J. Biol. Chem. 253, 4143--4152) has been subjected to isoelectric focusing. When monomer samples, shown to be homogeneous by both ion-exchange and molecular sieve chromatography, were analyzed by this technique, three subspecies were produced, which were similar in amino acid composition and specific CO2 hydratase activity. In addition to having either monomer or dimer status, the subspecies differed in the extent of oxidation of their sulhydryl groups and in their isoelectric pH values (9.3, 8.8, and 8.4, respectively). Also, the presence of dithiothreitol will affect their relative concentrations. These subforms are therefore designated as pseudoisoenzymes and are considered to be neither genetically nor functionally separate enzyme species.


A search for the function of human carbonic anhydrase B. Because of the very high activity and abundance of human red cell carbonic anhydrase C (carbamate hydrolase, EC 4.2.1.1), it seemed likely that the second isozyme, B, might not be essential for CO2 metabolism. It was then found that physiological concentrations of Cl- inhibited catalysis of CO2 hydration by the B enzyme (but not by type C), suggesting further that type B does not function in vivo as a carbonic anhydrase. The versatility of the catalytic activity of carbonic anhydrase for a number of 'artificial' substrates suggested that enzyme B may be utilized in reactions of intermediary metabolism. A number of hydration, dehydration, decarboxylation, kinase, and phosphatase systems were tested to determine a possible physiological function for the enzyme. Results with eighteen possible substrates were negative and the possibility is discussed that mammalian carbonic anhydrase B is an evolutionary accident.


1H NMR study of the conversion of 13(S)-hydroperoxy linoleic acid by soya bean lipoxygenase I. The conversion of 13(S)-hydroperoxy linoleic acid by lipoxygenase I at 298 K was monitored by 1H NMR and ultraviolet absorption spectroscopy. The rate constant for the conversion of the hydroperoxide, k = 45.8 +/- 7.5 M-1 . s-1, depends on the concentrations of both enzyme and hydroperoxide. This constant is not affected by O2, nor by solvent isotope effects.


The mechanism of activation of human prothrombin by an activator isolated from Dispholidus typus venom. Purified human prothrombin was activated, both in the absence and in the presence of thrombin inhibitors (diisopropylfluorophosphate or hirudin), by a coagulant principle isolated from Dispholidus typus venom. The process of activation was monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis. In the absence of thrombin inhibitor, prolonged incubation of prothrombin with the purified venom yielded thrombin, fragment 1 (F 1) and fragment 2 (F 2). In the presence of diisopropylfluorophosphate, which in the experimental conditions used inhibited only partially the thrombin generated activity, products obtained upon activation of prothrombin by venom were F 1 and a two-chain, disulfide-bridged protein of 58 000 daltons called meizothrombin (des F 1). In the presence of hirudin, which fully inhibited thrombin generated activity, prothrombin activation by the venom did not liberate any fragment, but prothrombin was converted to a derivative composed of two disulfide-bridged polypeptide chains of 48 000 and 37 000 daltons, called meizothrombin. These results are similar to those reported by others when studying the process of prothrombin activation by Echis carinatus venom and allow to conclude that Dispholidus typus venom cleaves a bond linking the A and B chains of thrombin, converting prothrombin into meizothrombin. This enzyme is then responsible for the cleavage of the bond linking F 1 and F 2 and the bond linking F2 the A chain of thrombin.


Thyroxine-protein interactions. Binding constants for interaction of thyroxine analogues with the thyroxine binding site on human thyroxine-binding globulin. The binding constants for interaction of various thryoxine analogues with the thyroxine binding site on human thyroxine-binding globulin have been determined. Equilibrium dialysis, at pH 7.4 and 37 degrees C, was used to measure the competitive effects of different iodothyronine compounds on the binding of 125I-labeled thyroxine to highly purified thyroxine-binding globulin. Relative to L-thyroxine, K = 6 . 10(9) M-1, the association constants of some important analogues were D-thyroxine, 1.04 . 10(9) M-1, 3,5-diiodo-3'-isopropyl-L-thyronine, 4.9 . 10(8) M-1; L-triiodothyronine, 3.3 . 10(8) M-1, 3,3',5'-DL-triiodothyronine (reverse triiodothyronine), 3.1. 10(8) M-1; tetraiodothyropropionic acid, 2.7 . 10(8) M-1; tetraiodothyroacetic acid, 2.6 . 10(8) M-1; 3', 5'- diiodo-DL-thyronine, 8.3 . 10(7) M-1; and 3,5-diiodo-DL-thyronine, 7.1 . 10(7) M-1. Calculation of the deltaG0 values for binding of the analogues indicates that a major contribution to the free energy favoring binding is made by the alanine side chain of thyroxine. A change in configuration of the alpha-amino group from the L to D form causes an unfavorable change of 1 kcal/mol in the free energy of binding. Removal of the alpha-amino group as in tetraiodothyropropionic acid causes an unfavorable change of 1.9 kcal/mol in the free energy of binding. With regard to ring substituents, the results indicate that the two inner 3,5-iodines make about the same contribution to binding as the two outer 3', 5'-iodines.


Amino acid sequence of the human myeloma lambda chain Win. The complete amino acid sequence of the variable region of a human myeloma immunoglobulin light chain (Win) has been determined. The sequence of the constant region has been verified by compositional analysis of its tryptic peptides. The amino acid sequence of the light chain Win corresponds to sub group II and shows no unusual amino acid replacements in its constant or variable regions when compared to other human gamma chains.


Regulation of lactate dehydrogenase synthesis in Bacillus subtilis. The regulation of lactate dehydrogenase in Bacillus subtilis was determined under a variety of growth conditions and in mutants blocked in the citric acid cycle. The synthesis of lactate dehydrogenase increased sharply concomitantly upon the exhaustion of glucose from the medium and the onset of the stationary phase. The synthesis of lactate dehydrogenase may be under catabolite repression control. Studies with mutants blocked in the citric acid cycle showed that lactate dehydrogenase is regulated independently of either the oxidative or reductase branches of the cycle. Certain citric acid cycle mutants, e.g., aconitase or succinate dehydrogenase, exhibited very low levels of lactate dehydrogenase while others, e.g., malate dehydrogenase or isocitrate dehydrogenase, showed normal levels. A stage O sporulation mutant expressed levels of lactate dehydrogenase more than one thousand-fold higher than the low group of citric acid cycle mutants. The induction of lactate dehydrogenase was shown to be independent of the accumulation of its substrate, pyruvate.


On the activities of glycogen phosphorylase and glycogen synthase in the liver of the rat. A procedure was developed for determination of glycogen synthase and phosphorylase activities in liver after various in vivo physiological treatments. Liver samples were obtained from anaesthetised rats by freeze-clamping in situ. Other procedures were shown to stimulate the activity of phosphorylase and depress the activity of glycogen in the liver. The direction of glycogen metabolism appears to be regulated by the relative proportions of the two enzymes, as shown by a strong positive correlation between total activities and active forms of phosphorylase and synthase. The enzyme activities responded as expected to stimuli such as insulin and glucose, which depressed phosphorylase and increased synthase activity, and glucagon, which increased phosphorylase and decreased synthase activity. In fasted animals approximately 50% of each enzyme was in the active form, which suggests the existence of a potential futile cycle for glycogen metabolism. The role for such a cycle in the regulation of glycogen synthesis and degradation is discussed.


[In vitro study of glucuronoconjugation of the toxin from Penicillium roqueforti (P.R.T.) and its metabolites]. Rat liver microsome UDP-glucuronyltransferase and labelled UDP-glucuronic acid were incubated either with P.R.T. or the compounds obtained by the in vitro metabolism of the toxin. Under the same conditions, labelled P.R.T. or its labelled metabolites were incubated with UDP glucuronyltransferase. Radioactive metabolites were produced with Eremofortin C and Eremofortin C alcohol and in each case, were identified as the corresponding beta-glucuronide conjugate. No measurable glucuronidation of P.R.T. or P.R.T. alcohol was observed. The results outlined in this paper show a good correlation between the biological effects and the ability of forming a glucuronide conjugate.


[Reaction between tetranitromethane and deoxyribonucleoproteins]. A comparative kinetic study has been made of tetranitromethane nitration of tyrosine residues in deoxyribonucleoprotein preparation (DNP) treated with EDTA and/or UV light at lambda=253.7 nm, as well as obtained by enzymatic digestion of nucleosomes. UV-light-induced protein-DNA linkages stabilize the structure of the preparation, whereas the action of chelate agents causes DNP-200 angstrom leads to DNP-100 angstrom transfer. Comparison of kinetic data and the results of the amino acid analysis of individual fractions of nitrated histones allowed to conclude that the differences observed between the degree and the rate of nitration are due to internucleosomic interactions which form the supernucleosomic structure of DNP.


[The role of a protonmotive force in genetic transformation of Bacillus subtilis]. The hypothesis on the role of protonmotive force in the transport of DNA through the membrane of Bac. subtilis cell during initial stages of genetic transformation was tested. A genetic transformation of arsenate-treated cells was observed. Treatment of cells by the protonophorous uncoupler of oxidative phosphorylation-carbonylcyanide dichlorophenyl--hydrazone-led to the inhibition of initial stages of genetic transformation having no significant effect on the level of intracellular ATP concentration and on the viability of cells. The dissipation of protonmotive force by means of K+ and H+ fluxes catalyzed by valinomycin and nigericin also caused the inhibition of initial stages of genetic transformation. The inhibitory effect of cationic penetrant tetraphenyl phosphonium was observed, the effect being potentiated by low concentrations of anionic penetrant phenyldicarbaundecaborate. The value of the membrane potential in the energized valinomycin-treated cells calculated from the distribution of K+ was within the range of 70--100 mV (inside minus). These results support the conception that a protonmotive force drives DNA transport through the membrane of Bac. subtilis cells.


[Enzymes of glycogen metabolism in chick embryo liver]. At all stages of ontogenesis glycogen phosphorylase (EC 2.4.1.1) from liver chick embryos in represented by an isoenzyme whose properties are close to those of isoenzyme IL or F. Total enzyme activity (a+b forms) from the 8th day of development up to hatching gradually increases 1.5-fold, a practically complete activation of enzyme being observed by the end of embryogenesis. Phosphorylase b possesses high catalytic activity in the presence of 1 mM AMP and it activated by protamine and 0.2 M Na2SO4. Glycogen synthetase (EC 2.4.1.11) has a constant Km(UDFG) value during ontogenesis. This value is about 5.10(-4) M in the presence of 10 mM glucose-6-phosphate, both for I- and D-forms of enzyme. The total enzyme activity reaches its maximum on the 17th postembryonic day and is decreased more than 6-fold thereafter. In the course of embryogenesis the I/D ratio is increased from 0.2 on the 8th day of development up to 0,45 during extensive accumulation of glycogen and falls down to 0.33 before hatching. Glycogen biosynthesis in embryonic liver is wellcorrelated with the increase in the I/D ratio, i.e. the increase of the active form of enzyme. The proportion of granular glycogen in embryonic liver is increased from 15% up to 90% of total glycogen content between the 8th and 14th days of development. The activity of glycogen synthetase contained in granular glycogen is increased from 40% in the 8-day-old embryos up to 90% in the 18-day-old ones. The activity of phosphorylase is found in granular glycogen only on the 12th day of embryogenesis and reaches its maximum (80% of total enzyme activity) only on the 19th days of development. It is concluded that in  the adult chicken liver the embronic enzymes--glycogen phosphorylase and glycogen synthetase--are retained.


Size increase and form allometry during evolution of ciliate species in the genera Colpoda and Tillina (Ciliophora: Colpodida). With the absence of a fossil record to provide direct evidence of evolutionary relationships, many researchers have inferred by circular reasoning that body size increase and form allometry have occurred during the evolution of ciliated protozoa. This study establishes ancestor-descendant relationships among five species of the related genera Colpoda and Tillina using ultrastructural characteristics which are not apparently dependent on size and form. This independent test suggests that phylogenetic size increase and phylogenetic form allometry have occurred during the evolution of these ciliates. Interrelationships of body size increase, increase in number of cortical organelles, and form allometry are discussed in reference to the divergence of these ciliate species.


[Effect of dimethyl sulfoxide on transformation of B. subtilis in vitro]. Bacillus subtilis transformation was conducted in the presence of dimethylsulfoxide and polyethyleneglycol. B. subtilis transformation was most frequent under the effect of 0.1% dimethylsulfoxide and was not altered significantly by polyethyleneglycol. As suggested, an increase of the transformation frequency was associated with the change of the membrane permeability under the influence of dimethylsulfoxide of with the altered activity of the membrane DNase.


[Changes in the surface morphology of normal and transformed mouse fibroblasts following disruption of cell-substrate adhesion]. By means of scanning electron microscopy surface morphology of cultured normal mouse embryo fibroblasts (MEF) and transformed mouse fibroblasts of L strain was studied in the course of alteration of cell-substrate adhesion with proteases, EDTA and urea. The morphology of cell rounding induced by the above agents in MEF and L cells was almost independent on the type of the agent. The rounding of MEF proceeded through three stages and was accompanied by substantial changes of cell surface relief. L cells lacked the intermediate stage (formation of thick processes) during their rounding which proceeded without any changes of cell surface relief. It is suggested that the observed differences are related to the poorer development of the lamelloplasm and microfilaments bundles in the transformed cells ascompared to the normal ones.


Fetal hemoglobin (HbF) synthesis in baboons, Papio cynocephalus. Analysis of fetal and adult hemoglobin synthesis during fetal development. Fetal hemoglobin (HbF) and adult hemoglobin (HbA) synthesis was studied in fetal baboons, Papio cynocephalus, to determine the normal pattern of hemoglobin production during fetal development. Fetuses ranging from 53 to 180 days gestation (term gestation 184 days) were used. Erythroid cells were incubated with 3H-L-leucine, and the rates of globin chain synthesis and the distribution of radioactivity into hemoglobin intermediates and completed hemoglobin molecules were determined. Gamma chain synthesis accounted for approximately 97% of the total nonalpha chain synthesis up to 140 days gestation; beta chain synthesis accounted for the remainder. After 140 days gestation, approximately equal quantities of gamma and beta chain were synthesized in the bone marrow. Prior to 140 days gestation, total alpha chain synthesis was 30% greater than total non-alpha chain synthesis, while there was balanced chain synthesis after 140 days gestation. During the period of excess alpha chain synthesis, fetal erythrocytes contained a large pool of alpha-hemoglobin (alpha chain with heme attached) molecules uncombined with beta or gamma chains. In view of the possibility that alpha chains may have a lower affinity for gamma chains than beta chains, excess alpha chain synthesis may be required to maintain low levels of free gamma chains.


Two-stage (initiation-promotion) carcinogenesis in vivo and in vitro. After a short review on the two-stage (initiation and promotion) chemical carcinogenesis in vivo, the conditions required for its demonstration have been reported with several experimental data, showing the role of two distince agents, the initiator and the promotor. The same phenomenon has also been demonstrated with cell cultures in vitro. Rat embryo cells treated with an initiator and then submitted to the continuous action of a promotor have been transformed more rapidly than the untreated cells. Transformation has been tested essentially by the tumorigenicity of the cells. The results indicate that tissue culture cells are an appropriate system to investigate the mechanism of two-stage chemical carcinogenesis and allow the detection of the initiating and promoting potentialities of various compounds.


Transmaternal modification of the Berenblum/Mottram experiment in mice. The classical 2-stage carcinogenesis experiment has been modified in that the carcinogen DMBA was applied to pregnant mother animals at different dose levels and different time intervals during pregnancy. Promotion with the phorbol ester TPA was performed as usual by application of the promoter on the back skin of mice of the F-1 generation. It can be shown that it is possible to initiate fetal epidermal cells by transmaternal route and to promote them to produce visible skin tumors post natum. Tumor promotion by TPA is not restricted to the epidermis, since a broad spectrum of tumors of internal organs can be demonstrated in the initiated and promoted animals. This more general promoting activity of TPA--which implies its absorption and distribution throughout the whole body--has not yet been described. In addition, especially sensitive phases during fetal life with regard to initiation of cells by the carcinogen can be demonstrated. These phases comprise the last third of pregnancy and coincide with a high proliferative activity and the onset of differentiation of the fetal epidermis. The results emphasize the important role of prenatal carcinogenesis and demonstrate the increased risk also to the human organism by either prenatal initiation or postnatal promotion.


Characteristics of chemically transformed mouse epidermal cells in vitro and in vivo. Primary cultures of epidermal cells from newborn mouse skin have been established. These cultures proliferate and express typical epidermal functions for limited time in vitro. The cultured cells keratinize and exhibit strong reactivity with an epidermis specific membrane antiserum. These cells can be transformed with DMBA either in standard cultures or more easily in cultures using 3T3 feeder cells. Transformed cells have increased proliferation rate. They do not pile up or form multilayered cultures like normal counterparts. They also keratinize less. When transformed cells were grown in the air on collagen gels they formed irregular clumps with central (horn-pearl) keratinization whereas normal controls formed stratified structures. Transofmred cells exhibited reduced reactivity with tissue specific membrane antiserum. There was masking or rearrangement of tissue specific membrane antigens with simultaneous exposition of new fetal-like antigens. Their reactivity with histocompatibility antisera was only slightly reduced. Transformed cells had both numerical and structural chromosome aberrations. They grew in soft agar and they induced tumors upon transplantation in the convenient host. When transplanted as cultures (on collagen) or as suspensions into the host, transformed cells were able to form epithelial cell cords invading the underlying mesenchyme with histological aspect typical of carcinoma. Cell cultures derived from in vivo DMBA induced tumors behaved like in vitro transformed cells.


Arsenic excretion by monkeys dosed with arsenic-containing fish or with inorganic arsenic. Four adult female Cynomolagus monkeys were given single oral dose of fish arsenic. 67% of the arsenic was excreted with the urine and 10% with the feces, essentially within 5 days post exposure. The same monkeys were later dosed with inorganic arsenic and 76% of it was excreted with the urine, and practically nil with the feces.


Aspirin, protein transacetylation and inhibition of prostaglandin synthetase in the kidney. 1 The effect of aspirin on the kidney has been investigated in mice and rabbits. [Acetyl-(14)C]-aspirin was administered intraperitoneally in doses ranging from subtherapeutic to toxic. The degree of acetylation of protein was determined by the radioactivity remaining on protein precipitates of renal cortex and medulla after sequential washing designed to remove non-covalently bound material. Controls were established, by the use of [carboxyl-(14)C]-aspirin.2 The acetyl-(14)C residue was bound to renal proteins in a linear manner in increasing amounts with increasing dosage up to 100 mg/kg. The [carboxyl-(14)C]-aspirin was not bound and thus the salicylate portion of the molecule was not bound covalently to the renal protein. The time course of the acetylation was rapid, consistent with the rate of aspirin absorption. The disappearance of acetylated protein was slow, with a T(1/2) of 112.5 h in the renal cortex, and 129.5 h in the renal medulla.3 Differential centrifugation, Sephadex chromatography and gel electrophoresis were carried out on tissue homogenates to determine the site of acetylation. The acetylation was greatest in the microsomal fraction, although all protein fractions showed some degree of acetylation.4 The prostaglandin synthetase activity of a particulate preparation from rabbit kidney was determined by a spectrophotometric assay of malondialdehyde formation. Aspirin (10 mg/kg, i.v.) significantly inhibited prostaglandin synthetase in the renal cortex and medulla.5 Aspirin and renal proteins undergo a transacetylation reaction resulting in stable acetylated protein, with acetylation being greatest in the microsomal fraction. Aspirin has been shown to inhibit prostaglandin synthetase and this could lead to functional impairment of the tissue.


Postoperative external small bowel fistulas: a study of a consecutive series of patients treated with intravenous hyperalimentation. A consecutive series of 25 patients who developed external small bowel fistula within 2 weeks of abdominal surgery is described. Half the patients had a primary diagnosis of inflammatory bowel disease and the fistula arose usually as a result of direct trauma to the bowel or the breakdown of an anastomosis. All the patients were treated conservatively with total bowel rest and intravenous hyperalimentation. In 15 (60 per cent) spontaneous fistula closure occurred, in an average period of 32 days. In 8 patients the fistula failed to close and surgery was performed, but was effective in only 3 cases. Thus the fistula eventually closed in 18 patients. Five patients died, all from intra-abdominal sepsis. Of the 8 patients with a primary diagnosis of Crohn's disease, 3 died, 2 have a persistent fistula, 1 has a permanent ileostomy and spontaneous closure occurred in only 2.


Changing patterns of resource allocation in a London teaching district. The health plans of the Tower Hamlets district management team were studied to determine what effects the report of the Resource Allocation Working Party and the White Paper "Priorities in the Health and Social Services" have had on resource allocation in a teaching district. The study showed that at present acute services are allocated a greater proportion of the district budget than occurs nationally, while geriatrics, mental health, and community services receive proportionately less. In the next three years spending on acute services is expected to decrease, while spending on geriatric facilities and community services will increase. Nevertheless, cuts in acute services will take place mainly through a reduction in the number of beds serving a community function, concentrating all acute services in the teaching hospital. Services to the district might be better maintained by creating a community hospital to meet the needs of patients who would otherwise need to be accommodated in acute beds with unnecessarily expensive support services.


Clinical trial experience by simulation: a workshop report. A new computer program for experimental design simulation was used in a three-day postgraduate workshop on clinical trials. Participants were given information on a fictitious new drug and asked to design a trial to evaluate it. On the first day they reviewed the data and designed a protocol, and results for these specifications were generated on the computer. The second day was spent analysing results, and on the last day the findings of the different studies were presented at a symposium. Flexibility of the program ensured a high degree of realism in all trials and emphasised the influence of design on results and conclusions.


Intravenous insulin infusion in diabetic emergencies. Continuous intravenous insulin and dextrose infusions were used in managing various diabetic emergencies. Standard and constant rates of insulin and dextrose infusion resulted in satisfactory control of blood glucose concentrations during labour, after major surgery, and in patients recovering from ketoacidosis (average insulin infusion rates 1, 2, and 3 U/h respectively). Higher infusion rates were used to correct or prevent ketoacidosis in pregnant diabetic women who had received steroids and sympathomimetic agents. The infusion method is simple, reliable, and flexible, and may help to simplify management of diverse types of diabetic emergencies.


Hypothermia: a complication of diabetic ketoacidosis. During 1969-77, 20 episodes of severe hypothermia occurred in 19 diabetic patients in Nottingham. Thirteen were associated with ketotic hyperosmolar coma, two with lactic acidosis, and one with hypoglycaemia, while in four there was no loss of diabetic control. Ketoacidosis accounted for 11.8% of all admissions for severe accidental hypothermia and was a commoner cause than hypothyroidism (8%). Patients with ketoacidosis were younger and developed hypothermia as often during the summer as during the winter. The metabolic disturbance was characteristic, with severe acidosis (mean pH 7.04), a high blood glucose concentration (mean 56.6 mmol/l; 1020 mg/100 ml), and high plasma osmolality (mean 379.7 mmol (mosmol)/kg). Eight of the 13 episodes proved fatal. Hypothermia may aggravate ketoacidosis and complicate treatment and should be sought in all patients with severe diabetic coma.


The luteinizing hormone-releasing hormone (LH-RH) neuronal networks of the guinea pig brain. I. Intra- and extra-hypothalamic projections. In the guinea pig brain, LH-RH-containing cell bodies are located not only within the classical hypophysiotrophic area but also in the medial preoptic area, septum and olfactory tubercle. LH-RH fiber tracts project not only to the primary portal plexus in the median eminence but also throughout the limbic forebrain and limbic midbrain regions. Using radiofrequency lesions in different brain regions, the projections of LH-RH cell bodies were determined. Cells in the medial preoptic area project ot the organum vasculosum of the lamina terminalis (OVLT), the suprachiasmatic nucleus, the mammillary body complex and the ventral tegmental area. LH-RH neurons in both the medial septal nucleus and medial preoptic area project via the stria medullaris to the medial habenular nucleus and from there via the fasciculus retroflexus to the interpeduncular nucleus of the midbrain. Other LH-RH neurons in the medial septal nucleus, nucleus of the diagonal band of Broca and olfactory tubercle are congregated in small clusters around large blood vessels which penetrate into this area, and they do not appear to send axons outside their immediate vicinity. The types of LH-RH axonal terminations and the roles of these peptide-containing neurons are discussed.


Operant control of pyramidal tract neurons: the role of spinal dorsal columns. A monkey was trained to control the firing patterns of precentral pyramidal tract neurons. The operant task was for the monkey to produce consecutive interspike intervals (ISI) within a requisite range, or target. The mean time off-target (error) is used to quantify the accuracy of control the monkey could assert over each PTN. Following partial destruction of the dorsal funiculi the number of PTNs driven by peripheral stimuli greatly decreased. Those PTNs which remained responsive to peripheral stimuli were as accurately controlled as those tested before column section, whereas, those PTNs unresponsive to peripheral stimuli were significantly less accurately controlled. The conclusion is that the monkey relies heavily upon proprioceptive feedback to operantly control precentral PTNs.


Effect of amino-oxyacetic acid (AOAA) on focal penicillin seizures. The biochemical and behavioral effects of the anticonvulsant amino-oxyacetic acid (AOAA) have been studied in a model of focal penicillin seizures in rats. At 20 mg/Kg AOAA treatment results in a progressive 11 fold increase in GABA levels in cortex over three hours. There is a decrease in aspartate, ketoglutarate, alanine and glutamine, and an initial decrease followed by an increase in pyruvate and glutamate. These results reflect a functional inhibition of several B-6 dependent aminotransferase enzymes. When rats are pretreated 30 min before the onset of focal penicillin seizures there is a 60% reduction in the number of discharges and a 34% reduction in seizure duration. Pretreatment beyond 75 min results in progressively less anticonvulsant effect, such that seizures eventually become more severe than control. There is an increase in the number and duration of discharges, seizure spikes become complex, and tonic-clonic events develop. Penicillin seizures do not cause a change in levels of GABA, but result in a decrease in glutamate within the focus. AOAA pretreatment initially prevents this decrease in glutamate but later accentuates it. The biochemical effects of AOAA are complex, but biphasic anticonvulsant properties coincide in time with a change in the balance of excitatory and inhibitory amino acids in the seizure focus.


Oculomotor nucleus afferents in the monkey demonstrated with horseradish peroxidase. The afferents to the oculomotor nucleus were examined in the macaque monkey by means of horseradish peroxidase. The demonstration of afferents from the contralateral interstitial nucleus of Cajal, and from the nearby rostral interstitial nucleus of the medial longitudinal fasciculus, predominantly ipsilateral to the injection site, confirms the significant role of these two regions as premotor structures. HRP label in the pretectal area was strictly confined to the olivary nucleus and the corresponding pathway seems to concern the Edinger-Westphal nucleus in particular. A purely crossed internuclear pathway emanating from the abducens nucleus could be established, and evidence for the presence of intranuclear connections within the oculomotor complex itself was found. Ipsilateral afferents from the superior vestibular nucleus and bilateral connections from the medial vestibular nucleus and the y-group were prominent. A projection, showing some rostrocaudal organization within the oculomotor nucleus, arises from the ipsilateral perihypoglossal complex, and other afferents originated from the underlying medullary reticular formation. No evidence was found for the existence of afferents from the paramedian pontine reticular formation, the nucleus Darkschewitsch, the nuclei of the posterior commissure, the lateral tegmentum and the dentate nucleus.


Demonstration of a specific localization of carbonic anhydrase C in the glial cells of rat CNS by an immunohistochemical method. The localization of carbonic anhydrase C isoenzyme in the central nervous system (CNS) of the rat has been investigated using the indirect immunoperoxidase technique, at both optic and electron microscopic levels. Evidence is presented for a specific localization of the enzyme in the cytoplasm of the oligodendrocytes and astrocytes. Myelinated fibers show a weak staining. The positive reaction is restricted to the cytoplasmic areas of the myelin sheath and does not appear in the compact myelin. Neuronal cell bodies do not stain at all. A strong positive reaction to the antiserum was also observed in the choroid plexus.


Hexamethylmelamine in alkylating agent-resistant ovarian carcinoma. Twenty-one patients with advanced ovarian cancer proven resistant to standard alkylating chemotherapy were evaluated in a prospective study of Hexamethylmelamine. All but three patients received 8 mg/kg daily with dose modifications appropriate to the degree of toxicity; the latter three patients were treated on an intermittent 14 day course of 8 mg/kg daily. Six patients (28%) experienced an objective response with a median duration of 2.5 months (range 1--5 months). The toxic effects requiring dose modification were gastrointestinal in 57%, hematologic in 62%, and neurologic in 28.5%. An average of 60.5% of the total projected dose was tolerated for a median of 44 days (range 7--113). Twelve patients (57%) experienced severe toxicity requiring discontinuation of therapy. It is apparent from this study that Hexamethylmelamine is an active agent in ovarian cancer and is not invariably cross-resistant to conventional alkylating agent therapy. These two characteristics make it an attractive agent for use in combination. Its toxicity in previously treated patients in the schedules used here, however, is substantial.


Preventive central nervous system irradiation in children with acute nonlymphocytic leukemia. In this study of children with acute nonlymphocytic leukemia an attempt was made to prevent central nervous system relapse and to determine whether this therapy, coupled with multiagent chemotherapy, would be successful in prolonging durations of complete remission. Central nervous system relapses were prevented by irradiation, although patients who received this therapy did no better than those who did not receive irradiation. A small group of patients received irradiation to the liver and spleen, but this modality also failed to improve the duration of remission. Control of extramedullary leukemia, in this study, failed to improve remission duration because bone marrow relapse was not prevented or delayed. It is unlikely that focal therapy will have a significant impact in acute nonlymphocytic leukemia until longer marrow remissions are achieved.


The role of radiation therapy in the treatment of pediatric non-Hodgkin's lymphomas. Between 1971 and 1976, 64 patients less than 18 years of age with non-Hodgkin's lymphoma were treated at Boston's Children's Hospital Medical Center-Joint Center for Radiation Therapy. A multimodality approach was used, consisting of radiation therapy (3500--4500 rad), surgery, and chemotherapy. Since 1973, all patients have received a regimen initially comprising Adriamycin, Prednisone, 6-Mercaptopurine, Vincristine, and L-Asparaginase. Methotrexate was substituted for Adriamycin following a cumulative total dose of 450 mg/m2. The 5-year actuarial survival for all patients was 61% while relapse-free survival was 54%. The actuarial and relapse-free survival for patients presenting with localized disease was 75% and 72%, respectively. Median follow-up was 40 months and all relapses occurred within 24 months of initial therapy. A multidisicplinary approach, such as the current regimen, offers a good prognosis for this disease.


Radiation therapy for germinal tumors of the testes. 79 patients with germinal tumors of the testes were treated at the University of Maryland Radiation Therapy Clinic between January 1, 1957 and December 31, 1973. 32 of 33 patients with stage I seminoma had disease controlled for a minimum of three years for a 96.9% 3 year disease-free determinant survival. 4 of 4 patients with stage I carcinoma were treated, two after negative node dissection, and all survived. 15 of 21 patients with stage II seminoma survived for 3 year disease-free Berkson-Gage actuarial survival of 68.5%, 9 of 21 stage II carcinomas, 18 of whom had had positive node dis-sections, survived for a 3 year disease-free Berkson-Gage actuarial survival of 42.8%. Benefit of adjuvant irradiation of the mediastinum and supraclavicular regions is demonstrated for stage II seminomas where 7/8 patients receiving prophylaxis demonstrated a 100% 3 year disease-free Berkson-Gage actuarial survival as opposed to the 50.4% in the 7/13 patients surviving without adjuvant therapy. This benefit is reaffirmed, although more tenuously, by the 71.4% 3 year disease-free Berkson-Gage actuarial survival demonstrated by the 5 of 7 stage II carcinoma patients receiving adjuvant therapy as opposed to the 28.2% demonstrated by the 4 of 14 patients surviving who did not receive adjuvant therapy.


Carcinoma of the vagina.

Carcinoma of the vagina. Twenty-two patients with Stage I through IV primary vaginal squamous cell carcinomas treated for cure with radiation therapy are reviewed, with particular emphasis on the relationship of dose to complications and local control. All but 2 patients received 4000 to 6000 rad whole pelvis irradiation plus at least one radium application. Local control was 91%, with an absolute 2-year disease-free survival of 82%. The degree of anaplasia was found to influence prognosis, with all local and distant failures resulting from high-grade lesions. The complication rate was modest, with no fistulae or serious bowel complications. An analysis of total dose (external plus radium) with respect to local failure and complications showed that no major complications occurred at a combined dose below 9000 rad. An analysis of the individual contributions of external irradiation and radium implants showed that all but one very minor complication occurred at a radium dose of 4000 rad or higher. From these data, overall treatment planning and total dose recommendations are made.


Acquisition of angiogenic capacity and neoplastic transformation in the rat mammary gland. The ability to induce formation of new vessels was tested in fragments of rat mammary tissue transplanted onto the rabbit iris and observed through the transparent cornea. Virgin, pregnant, and lactating glands showed an angiogenic capacity in about 5% of implants. In contrast mammary carcinomas induced angiogenesis in 75 to 100% of implants. Fragments of mammary gland previously treated with 7,12-dimethylbenz[alpha]anthracene of N-nitrosomethylurea but without histological evidence of neoplastic transformation showed an angiogenic response in about 5% of implants. The same low angiogenic response was detected in primary hyperplastic alveolar nodules. However, angiogenesis was observed 2 to 3 times more frequently in implants from hyperplastic outgrowths that acquired of continuous transplantability and showed a high degree of neoplastic transformation. These data on the rat mammary gland confirm previous findings on mouse mammary gland, indicating that: (a) neoplastic epithelium has a higher angiogenic capacity than does normal epithelium; and (b) hyperplastic epithelium at high risk of undergoing neoplastic transformation induces angiogenesis more frequently than does hyperplastic epithelium with low tumor potential.


Inhibition of DNA replicon initiation by 4-nitroquinoline 1-oxide, adriamycin, and ethyleneimine. The effects of three widely differing chemical carcinogens, 4-nitroquinoline 1-oxide, Adriamycin, and ethyleneimine, on DNA replication were studied by pulse labeling of DNA with [3H]thymidine and sedimentation analysis with alkaline sucrose gradients. At doses that reduced the rate of DNA synthesis to 30 to 60% of control values, only ethyleneimine produced damage that resulted in lower molecular weights of parental DNA. All three chemicals inhibited replicon initiation, but to differing extents. Inhibition of replicon initiation was the first clearly identified effect of 4-nitroquinoline 1-oxide and was the main cause of inhibition of DNA synthesis. Ethyleneimine caused severe inhibition of replicon initiation, but blocks to chain elongation also contributed significantly to the inhibition of overall DNA synthesis. Adriamycin affected replicon initiation to a small but significant extent; the primary cause of inhibition of DNA synthesis by this drug was a slowing of the rate of chain elongation. These results indicate that inhibition of replicon initiation is an important mechanism for the action of DNA-damaging agents in mammalian cells and strengthen the concept that control of DNA replication depends on the structural integrity of a chromosomal subunit that consists of several replicons.


Expression of Ricinus communis receptors on epithelial cells in oral carcinomas and oral wounds. The histological distribution of receptors for Ricinus communis Fraction 1 (RCA1) in oral carcinomas and in oral epithelial cells during wound healing has been studied by use of fluorescein-tagged RCA1. Biopsies from 15 human oral carcinomas and adjacent normal mucosa showed RCA1 receptors at the cell membranes in the basal and spinous layer of the normal epithelium, whereas receptors could not be demonstrated in invading islands of the tumors. In healing oral wounds from eight humans and three monkeys, RCA1 receptors were demonstrated both in normal epithelium adjacent to the wounds and in the epithelial outgrowth from the wound margin. Titrations, however, showed that the epithelial outgrowth reacted more weakly than did the normal adjacent epithelium. These results support previous in vitro studies showing changes in carbohydrate composition of moving normal cells and of malignant cells, a finding that may be of interest in relation to formation of metastases.


Ultrastructure of the different zones of the tectorial membrane. The tectorial membrane (t.m.) of mammals, which lies over the organ of Corti, is made up of an agglomerate of protofibrils of varying degrees of hydration. Two types of protofibrils are clearly distinguishable in the mouse t.m. While type-A protofibrils are straight and unbranched (thickness: 110A) demonstrating a periodic structure (period = 70 A), type-B protofibrils are branched and coiled (thickness: 150-200 A). These protofibrils could be systematically ordered according to the different t.m. zones. Type-A protofibrils predominate in the basal layer and in the entire middle zone, where they are interlaced with strongly hydrated type-B protofibrils. Weakly hydrated type-B protofibrils essentially make up the marginal zone (with the marginal net), the covering net, Hensen's stripe and the immediate contact layer with the limbus.


Isolation, structural studies and chemical synthesis of a 'palmitone lipid' from Corynebacterium diphtheriae. The isolation of a 'palmitone lipid' from Corynebacterium diphtheriae is described. The use of a temporary hydrophobic protecting group allows the obtaining of the lipid in free and pure form. Structural studies by chemical degradation and mass spectrometry allow one to propose structure Ic for this compound, namely 6-(2-tetradecyl 3-keto octadecanoyl)-alpha-D-trehalose. This structure was confirmed by chemical synthesis.


Total synthesis of ceramide trihexoside accumulating with Fabry's disease. We report the synthesis of natural ceramide trihexoside, viz. gal (alpha-1 leads to 4) gal (beta-1 leads to 4) gluc (beta-1 leads to 1) ceramide (XIII). It involves the Koenigs-Knorr reaction of the bromide II with the aglucon XIX, and of the chloride VII with the D-enantiomer of the ceramide ester XII. A positional isomer of XIII was obtained as a by-product. A novel technique in the Koenings-Knorr reaction is described.


Micelle and acid-soap formation of linoleic acid and 13-L-hydroperoxylinoleic acid being substrates of lipoxygenase-1. Surface tension measurements of linoleic acid solutions in 0.1 M sodiumborate buffer pH 10 at 23 degrees C showed that at increasing the linoleic acid concentration a sharp transition from monomers to micelles occurs at 167 micrometer. At pH 9 and 8 formation of acid-soap dimers from monomers starts at 60 micrometer and 21 micrometer respectively. The concentration range at which only monomers exist is therefore markedly reduced. For 13-L-hydroperoxylinoleic acid at pH 10 acid-soap formation still takes place, starting at approx. 220 micrometer. The total lipid concentration at which acid-soap or micelle formation starts in mixtures of linoleic acid and 13-L-hydroperoxylinoleic acid has been determined in relation to the molar ratio of both acids.


[Epidemiology of leishmaniasis in French Guiana in relation to the existence of a new species of echimyid rodent, Proechimys cuvieri sp. n]. The rediscovery and reexamination of the skull of the type of Proechimys guyannensis (E. Geoffroy; 1803) allows us to give more details about this species. It appears that a new sympatric species, P. cuvieri, has to be distinguished, which could have great implications for the leishmaniosis epidemiology in French Guiana.


[Action of carbaryl on the electroretinogram of the white mouse]. Carbaryl intoxication results in important modifications of the Mouse ERG, with an increase of the latency and a decrease of the amplitude, then a suppression of b-wave. This action is supposed to be provoked by the anticholinesterasic properties of carbamates, which result in a section of retinian synapses.


[Action of somatostatin and 3 analogs on external pancreatic secretion in the rat]. The effect os somatostatin (SS) and of its analogs D Trp 8-SS and Asn 5-SS was studied upon the external pancreatic secretion of the Rat after stimulation by 2 deoxy-D-glucose. The secretion of sodium and bicarbonate was similarly inhibited by all four peptides. The analog D Trp 8-SS was more effective in inhibiting pancreatic protein excretion.


[Plasma estradiol levels in quail during sexual development and after different estrogenic treatments]. The comparison between estradiol levels in peripheral plasma of developing quails and of estradiol benzoate treated quails (0.01, 0.02, 0.1 and 1 mg/daily/6 days) shows that only the dose of 0.01 mg reproduces the conditions of a physiological estrogen stimulation.


[Adrenal gland growth of the ovine fetus in the end of gestation: evolution of DNA and of membrane proteins]. The adrenal weight of the ovine fetus increases dramatically during late pregnancy. This increase is achieved in three periods: one of DNA duplication between two phases of cellular hypertrophy. During cellular hypertrophy an important biosynthesis of membrane proteins is observed. This development concerns essentially the adrenal cortex.


[Pseudomonas aeruginosa epidemiology in Egypt: study of 80 hospital strains isolated at Alexandria]. Eighty hospital strains of P. aeruginosa, isolated in Alexandria, were examined with the aid of three epidemiological markers (serotype, lysotype and O. N. P. G.-test). The most frequent serotypes in Alexandria are 0:11, 0:12, 0:2, 0:4, 0:6 et 0:1; this situation is different from those reported in European countries and in U. S. A. Sixty-five lysotypes were found and the strains of serotype 0:11 fall in fifteen lysotypes. One strain 0:12 was found O. N. P. G. positive.


[Speed of in vitro reassociation of gonadotropin subunits in a teleost fish, the carp (Cyprinus carpio L.)]. Purified gonadotropin subunits of a teleost Fish, the Carp, reassociated in vitro, according to a second order rate reaction. The rate constant was temperature dependent. Compared to ovine LH, the Q10 was about three times less and the rate constant much greater (160 times for instance at 20 degrees C). The possible physiological significance of these differences is underlined.