Pathogenesis of acute pancreatitis: pancreatic ductal-interstitial-vascular and lymphatic pathways. The generally accepted theory explaining the pathogenesis of pancreatic autodigestion (acute pancreatitis) is analysed. The pathways are discussed through which the content of the pancreatic duct system, the digestive enzymes, may escape to enter the acinar cells and the pancreatic interstices, together with the effect of the enzymes on the pancreatic vessels and thus on pancreatic blood flow. The potential consequences of vascular damage are described as well as the importance of pancreatic lymphatics in the transport of the escaped enzymes from the interstices. Although no definite answer can be given to the basic problem whether acute autodigestion of the pancreas is a result of some intra- or extra-acinar process, it is suggested that the intra- and extra-acinar processes are linked and are decisive in launching the pathological process.


Effect of immunosuppressive therapy on the serum glycoprotein levels in chronic aggressive hepatitis. The sera of 24 patients with chronic aggressive hepatitis receiving combined immunosuppressive therapy were studied for the concentrations of the carbohydrate components of glycoproteins and the IgG, IgA, IgM, alpha-2-macroglobulin, coeruloplasmin, beta-1-C-globulin and transferrin levels over a period of 2 years. Liver biopsy was performed repeatedly in 50% of the cases. On the evidence of the results, combined immunosuppressive treatment is regarded as apt to normalize the serum concentrations of IgG, IgA, IgM and to reduce those of alpha-2-macroglobulin and coeruloplasmin. Among the carbohydrate components of glycoproteins only the amount of hexose was reduced.


Carbohydrate metabolism in various types of hyperlipoproteinaemia. Insulin secretion has been studied after an oral glucose load in 81 patients with primary hyperlipoproteinaemia and 15 control sugjects. Glucose tolerance was reduced mainly in Type IV, and to a lesser extent in Type IIB hyperlipoproteinaemia. Insulin secretory response to glucose was reduced in Type V and heterogeneous in Type IV hyperlipoproteinaemia. No correlation was found between the triglyceride level and the rate of insulin secretion.


Serum lipids and lipoproteins in chronic liver disease. Serum cholesterol, triglyceride, total lipids and the lipoprotein pattern were studied in 169 cases chronic liver disease confirmed by biopsy. On the ground of the immunological and morphological results the patients were classified into five groups. In chronic persistent hepatitis no significant abnormality was found. In chronic aggressive hepatitis and in cirrhosis of the liver the serum cholesterol level was significantly reduced. In fatty infiltration of the liver the serum cholesterol, triglyceride and total lipid concentrations were significantly increased, as compared with the normal values and with the figures obtained in the cases of chronic inflammatory liver disease. In the cases of cirrhosis with additional diabetes the lipid values were likewise increased. In chronic aggressive hepatitis and in cirrhosis of the liver the levels of pre-beta and alpha lipoprotein were decreased, in fatty infiltration of the liver those of beta and pre-beta lipoprotein were increased.


Diagnostic evaluation of gastric analysis (test meal) in Hungary. On the basis of information received from 128 medical wards, the frequency of gastric analysis (test meal) used as a clinical diagnostic procedure, the spreading of the different methods and other problems connected with the examination are analysed. In about 3/4 of the institutions the test meal is performed in patients with all kinds of gastric complaint. This incidence decreases to 50% if the X-ray finding is positive. As acid-stimulants caffeine and a moderate dose of histamine are used most frequently. Of the data supplied by the test meal, the value for acid concentration (acidity) is registered by most of the examiners. The authors consider it as suprising that outdated methods such as a qualitative lactic acid assay (47%), isolation of Boas-Oppler's bacillus (25%) and of sarcinas (21%) are still in frequent use. The data are compared to the results of a similar survey made in England.


Regulatory hyperparathyroidism: the role of C-cell hyperplasia. Four cases with clinical and biochemical evidence of hyperparathyroidism are reported. The syndrome was due to DHT-intoxication of iatrogenic origin in the first case to histologically confirmed C-cell hyperplasia of the thyroid in the three others. The collective term "regulatory hyperparathyroidism" is porposed for the syndromes which, in distinction to secondary parathyroidisms, have a pathogenesis other than a negative calcium balance. Management of the syndrome is discussed with particular emphasis on the removal of the TCT-producing hyperplasia or adenoma, and the restoration of the electrolyte balance.


Heart block: histological and clinical features. The clinical types of A-V-block and the localization of the underlying lesion to the conduction pathway have been examined in 25 patients. On the evidence of the post mortem studies the lesions were of the following types: 1) Necrosis or fibrosis of ischaemic origin: 13 cases; 2) lesions of inflammatory character: 5 cases; 3) nonspecific lesions; 6 cases; 4) malignant infiltration: one case. Correlations between the site of lesion and the clinical type and pattern of block could be demonstrated in a large proportion of the cases. The characteristic features of the individual types of block and their localization are discussed and a review of the literature is given.


ECG changes in benzene, toluene and xylene poisoned rats. Electrical activity of the heart was studied in acute, subacute and chronic benzene, toluene and xylene poisoning by means of NEK-2 type apparatus in CFY male rats. In the early period of acute intoxication benzene and its methyl derivatives, injected either intraperitoneally or subcutaneously or administered by inhalation did not cause changes in the ECG. Intravenously injected toluene induced a rapid and permanent decrease in blood pressure, in higher doses respiratory paralysis. In acute inhalation toluene and xylene poisoning the anesthesia was followed by respiratory paralysis, bradyarrhythmia and asystole, while inhalation of benzene induced respiratory paralysis followed by ventricular fibrillation. Subacute poisoning with benzene, toluene and xylene caused disorders in repolarization and arrhythmia. The degree, incidence and development of these phenomena depended on the dose and the duration of exposure. The effect of benzene influencing ECG was appreciably more intensive than that of toluene or xylene. Owing to the high number of spontaneous ECG changes due to ageing, it is difficult to evaluate the effect of chronic exposure to the agents under investigation.


Modification of the biologic dose to normal tissue by daily fraction. Model for calculating normal tissue tolerance. A method to predict normal tissue injury is proposed that includes high daily doses and unusual times successfully by calculating a new value called BIR (Biologic Index of Reaction). BIR and NSD were calculated for various normal tissue reactions. With the aid of statistical correlation techniques it is found that the BIR model is better than the NSD model in predicting radiation myelopathy and vocal edema and as good as NSD in predicting rib fracture. Neither model predicts pericardial effusion. In no case were the results  of BIR inferior to those of NSD.


Anti-oestrogen therapy of advanced mammary carcinoma. Forty mg daily of tamoxifen (Nolvadex) was administered orally to 89 patients with advanced soft tissue mammary carcinoma. At two months 43 per cent of the patients responded. At 6, 12 and 18 months, 42, 35 and 32 per cent, respectively, were still responding to therapy. The side effects were limited, fatique being the most frequent complaint. Oestrogen dependent side effects were not encountered. The therapeutic effect of tamoxifen is similar to that of oestrogen, but the side effects are less.


Scanning electron microscopy of cells in the lymph of the human thoracic duct in advanced malignancies. At scanning electron microscopy of cells from the lymph of the thoracic duct, collected by cannulation of patients with malignant tumours of different types, lymphocytes and red blood cells were found, but few malignant cells. The latter came from patients with pulmonary carcinoma and mammary carcinoma with lung metastases. Various abnormal shapes of the red blood cells were observed, possibly due to extrinsic factors such as administered drugs of abnormal metabolites from the malignant disease. The lymphocytes had no consistent pathologic shape, but appeared to act as immunologic, active cells against the malignant cells in the thoracic lymph.


Radiation sensitizing effect of diamide on human cells cultivated in vitro. Human cells of line NHIK 3025 were irradiated suspended in growth medium (E2a) in absence and presence of diamide under aerobic and extremely hypoxic (less than 4 ppm O2) conditions. A sensitizing effect of diamide was found for doses exceeding 8 Gy (800 rad) on cells irradiated under extremely hypoxic conditions in presence of diamide of concentration 200 micrometer, whereas no significant effect was observed for 20 micrometer.


Histochemical and morphometrical changes in the adrenal glands in the course of circulatory failure. The histological and biochemical changes in adrenal function has been studied in 50 male cats with circulatory failure. The adrenal glands were examined by morphometric, histochemical and histoenzymological methods after a preliminarily study of the histoenzymological profile of the adrenal glands in intact animals. Five hours after hypotension induced by intravenous injection of trimethaphan and 5 hours after a single massive bleeding, a decrease in adrenal function was observed. Adrenal insufficiency especially of the glucocorticoid release in the course of acute circulatory failure can be considered as a peculiar adaptation of the organism to preserve its life. The demonstrated decrease of adrenal function in the first hours of hypoxia may help in planning hormonal therapy under suitable conditions.


Development of collagenous fibres in autologous and preserved homologous tendon grafts. Collagen synthesis has been studied in preserved autologous and homologous tendon grafts, applied in dogs. The stages of collagen morphogenesis observed in the synthetizing fibroblasts appearing in the decomposing original structure of the transplants as well as in the extracellular ground substance are described. The results suggest that autologous tendon grafts and homologous ones preserved by irradiation, stored in cold physiological saline and beta-propiolactone, are rebuilt within eight weeks; meanwhile the original collagenous fibre structure is decomposed. When other preserving procedures such as deep-freezing or gamma irradiation are applied recomposition of the graft is retarded or fails to occur; the result is a cicatrizing adhesion.


Selective sympathetic denervation induced by 6-hydroxy-dopamine in the small intestine. The structure of the nerve fibres in the intact and neurally isolated intestine of the cat was studied after 6-OHDA treatment by fluorescence and electron microscopy. A gradual disappearance of green fluorescence in the monoaminergic fibers was observed. Degeneration of nerve fibres containing 30--60 nm agranular and 30--60 nm densecore (granular) vesicles could be seen in all layers of the small intestine. Sporadically, degeneration of the presynaptic elements occurred also in the synapses of myenteric ganglia. A large number of degenerating fibres could be observed in close relation to blood vessels. These were thought to be monoaminergic because 6-OHDA selectively affected their terminals. In the chronically isolated small intestine degenerated nerve processes could not be observed following 6-OHDA treatment. It is concluded that the intrinsic nerve elements do not contain monoamines. Accordingly, the previously observed yellow fluorescence of the nerve elements of the isolated small intestine might be due to some other kind of neurotransmitter, possibly tryptamine.


Parenchymal destruction and arterial changes (haemodialysis intimal fibrosis) of glomerulonephritic kidneys in chronic intermittent haemodialysis. The kidneys of 13 chronic diffuse glomerulonephritis patients who had been subjected to chronic intermittent haemodialysis were compared with those of 6 patients treated in the conservative way. Their renal parenchyma was examined with a semiquantitative histological method and advanced complete destruction of the glomeruli was demonstrated in the dialysed cases. There was no perceptive difference in the distribution of the various grades of damage of the recognizable glomeruli and tubules. With the exception of the chronic lobular glomerulonephritis cases, a marked obliterative mucoid intimal proliferation ("haemodialysis-intimal-fibrosis") had developed in the dialysed glomerulonephritis patients. It affected the renal arteries of all size from the segmental to the interlobular arties. Haemodialysis-intimal-fibrosis did not develop in the additionally examined 13 dialysed chronic pyelonephritis cases nor in 8 such treated conservatively. The change seems to be characteristic, but not specific, for chronic diffuse glomerulonephritis treated by chronic intermittent haemodialysis.


Reversibility and interaction with hepatic regeneration of toluene induced liver injury. Acute and subacute toluene poisoning was induced in CFY rats. Routine histological, enzyme histochemical and electron microscopic investigations revealed that following discontinuation of exposure, the hepatic changes indicating an enhanced load on the detoxicating function (increased SDH activity, increase of mitochondria and smooth endoplasmic reticulum, decrease in Best carmine staining and PAS positivity) as well as degeneration (dilation of endoplasmic reticulum, accumulation positivity) as well as degeneration (dilation of endoplasmic reticulum, accumulation of autophagous vacuoles) show a rapid regression. The toxic effect of toluene and the functional load on the liver is thus reversible. In another series, toluene exposure was combined with partial hepatectomy. It has been established by routine histological, enzyme histochemical and electron microscopic techniques as well as by quantitative light and electron microscopic methods that the two interventions show a peculiar interaction: hepatic regeneration following partial hepatectomy inhibited the effect of toluene. On the other hand, the rate of regeneration was not influenced by toluene.


Myeloid metaplasia in the spleen after acute haemolysis. Copper sulphate, phenyl hydrazine and antiserum were administered to adult albino rats to produce equivalent haemolysis. Red pulp activity was studied for myeloid metaplasia after 24, 48 and 72 h, 5 and 7 days. Myeloid metaplasia was not related to the marginal zone proliferation. The activity rose rapidly with all the three agents and followed a similar curve despite the different patterns of haemolysis and erythrophagocytosis. Splenic erythrophagocytosis acts as a powerful feedback for splenic myeloid metaplasia. With copper, the release of reticulocytes and iron binding in haemoglobin is rapid as compared to the other agents, in spite of the initial lag phase, due to its effect on cellular metabolism.


Frequency of juxtaglomerular granulated cells in the mouse kidney. Studies of 23 untreated adult mouse kidneys revealed that in mouse kidney sections the frequency of juxtaglomerular granulated cells as compared to the glomeruli is 38.5 +/- 1.79%, the value for the JGI, 71.8 +/- 3.93. Following 100 glomeruli through complete serial sections prepared from a single mouse kidney, it was shown that in the cortex of the mouse kidney all juxtaglomerular apparatus related to the glomeruli contain renin-producing modified smooth muscle cells with granulated cytoplasm.


A glycoprotein precipitate in the blood vessels of kidneys stored in cold homologous blood. Dog kidneys were stored three hours at 4 degrees C in heparinized homologous blood. Their histological examination revealed the presence of a homogeneous precipitate in the glomerular capillaries and afferent and efferent arterioles, and histochemical analysis indicated that it consisted of glycoprotein. It is tentatively suggested that the substance is cryoglobulin, a conclusion supported by the disappearance of the deposit on reimplantation of the kidney.


Alterations of the Z-band in cardiac and skeletal muscles. The heart muscle may react to various hypoxic damaging effects (e.g.N2, CO, haemorrhagic shock, electroshock) by identical responses similarly as in the case of skeletal muscle damages. One of the early manifestations of the process is the alteration of the Z-band, which is considered pathological. The alterations of the Z-band region precede the changes of the mitochondrial and sacrotubular systems and might form the morphological basis of functional changes induced by hypoxic effects. The alterations observed are differentiated from the hypertrophy of the Z-band. In the development of the alterations of the Z-band the role of other factors (e.g. calcium metabolism, sacroplasmatic membrane changes) is emphasized.


Central core in the spinal grey matter.

Central core in the spinal grey matter. Golgi--Kopsch impregnated spinal cord sections of adult cats revealed a regular orientation of the dendrites in the intermediate region forming a circular or elliptical grey matter. The orientation of the dendrites is strictly transverse in the central core. A new interpretation of the spinal grey matter is proposed by viewing the dorsal, lateral and ventral horns (or columns) as crest-like appendages attached to the rod-like central core. A structural and possibly functional similarity between the central core and the brain stem reticular formation is emphasized.


The cytoarchitecture of the adrenal medulla in the rat. The disposition of adrenaline and noradrenaline storing cells in the adrenal medulla has been studied in the rat. It has been demonstrated that no morphological or functional reason exists for the follicular designation of clumps of chromaffin cells in the medulla. No significant difference was found between the outer and inner zone as regards the frequency of NA cells by morphometric and statistical methods. The cytoarchitectural differences between adrenaline- and noradrenaline-storing cell arrangement are described. Special features of the interrelation between noradrenaline storing cells, cortical cells and the connective tissue framework are stressed. The light and electron microscopic appearance of two types of cortical cells, assumed to represent different states of functional activity is described.


Intimate contact between human yolk sac endoderm and haemopoietic precursor cells. Although the localization of embryonic haemopoietic cells in the endodermal epithelium of the human yolk sac had been discovered several decades ago, the nature and significance of the localization were dubious, and it was supposed that light microscopic pictures represent technical artifacts. Our ultamicrographs now demonstrate that at 16--26 mm CR-length there is an intimate contact between yolk sac endoderm and free haemopoietic precursor cells: apart from interdigitations various types of intercellular contact could be discovered. These contacts, especially the gap-like ones with associated electron dense cytoplasmic areas speak for intercellular communication, i.e., the role of endoderm in early human embryonic haematopoiesis appears likely. The demonstrability of these contacts, however, does not mean that endodermal associations are indispensable for haemopoietic differentiation.


Blood supply of the rat hypothalamus. IV. Retrochiasmatic area, median eminence, arcuate nucleus. The vascular supply of the rat hypothalamo-hypophyseal complex was studied by aid of a double injection technique (two dyes different in colour injected successively). The vascular bed of the median eminence, the arcuate nucleus, and the retrochiasmatic area are intimately interconnected among themselves, but isolated from the vascular system of all other parts of the hypothalamus. The arterial supply of this ensemble of the medial basal hypothalamus is common from the hypophyseal arteries, via the primary plexus and the specific vascular loops of the median eminence. From here a separation of the vascular bed leads the blood flow towards the pituitary portal vessels on the one hand, and over a subependymal vascular network towards the arcuate nuclei on the other. The venous drainage of the arcuate nuclei and the retrochiasmatic area is secured by a special set of tuberal and retrochiasmatic veins. --The unity of the vascular bed of the median eminence, arcuate nucleus, retrochiasmatic area complex, with a blood flow largely in the direction indicated by this sequence, is of considerable importance for the interpretation of the results of experimental interference with this region.


Light and electron microscopic studies of the gastrocnemius muscle in peripheral occlusive vascular disease.


Rhinoplasty results.

Rhinoplasty results.


Rehabilitation of the voice after pharyngolaryngectomy and skin repair. I. Cineradiography. Eleven patients who had undergone pharyngolaryngectomy and who had poor or no speech were studied to find out why. An absent oesophageal stripping wave and the presence of a 'sump effect' are described. It is concluded that these are two positive anatomical factors mitigating against the acquisition of oesophageal speech in pharyngolaryngectomized patients.


A morphometric study of the laryngeal saccule and sinus. A morphometric study of 111 larynxs from both sexes was performed with the initial intention of assessing the incidence of asymptomatic laryngeal pouches. Certain basic measurements were made also to determine the dimensions of the laryngeal saccule and sinus relative to the height and width of the larynx in the two sexes. Asymptomatic pouches were found in 2 patients, one from each sex and there was no obvious difference in the height of the saccule relative to the height of the larynx in the two sexes. However, since the male larynx was deeper in the anteroposterior plane relative to its height, the male saccule tended to be relatively shallower than the female.


Late recurrence or radiation induced cancer of the larynx. Twenty five patients with squamous carcinoma of the glottis (T1a and T1b) had undergone successful irradiation. Many years later they developed a so-called  late recurrence. The following evidence shows that these 'late recurrences' are radiation induced: the interval of 5-18 years (mean 9.9 years) between the first and the second cancer correlates with the interval seen usually in radiation induced malignancy, recurrences generally appear during the first 2 years after irradiation, all the second cancers were found in the previously irradiated area, in patients treated by surgery only, late recurrences are extremely rare, histological examination clearly shows that the second carcinoma originates from the squamous epithelium and not from dormant nests in the deeper layers of the vocal cord. These factors should be taken into consideration when deciding between surgical or radiation therapy in younger patients with high life expectation.


Temporary threshold shift and the auditory evoked response. A preliminary report. A preliminary study was performed on 5 normal subjects to investigate and compare the reduction and recovery of the auditory evoked response (AER), and to compare this with the temporary threshold shift (TTS) that occurs with fatiguing sounds. The findings are discussed in relation to the basic nature of the AER.


The place of cytology in laryngeal diagnosis. A cytological procedure providing useful supplementary information in the diagnosis of laryngeal malignancy is described. Of 51 patients, 22 had complete correlation with histological examination in confirming a benign condition. Twenty patients had complete correlation in confirming a malignant lesion. Five patients showed cellular atypia in the presence of benign conditions. Three showed atypia, of which 2 were malignant, 1 being unstable histologically but with no evidence of frank malignancy. Ten patients had been irradiated previously; in all cases the distinction between residual disease and no evident disease was correctly diagnosed with respect to the histological appearance.


Cardiac arrest in otorhinolaryngology.

Cardiac arrest in otorhinolaryngology. A study has been carried out of 42 episodes of cardiac arrest related to 90 000 otorhinolaryngological operations. The basic pathology, factors precipitating the arrest, and the results of treatment are analysed. Clinical areas presenting special problems are the surgery of cancer of the postcricoid area, deliberate hypotension, anaesthesia for laryngoscopy and respiratory obstruction.


Facial spasm.

Facial spasm. There are many varieties of abnormal facial movement. This paper discusses hemifacial spasm and blepharospasm in particular. A technique found useful for the symptomatic relief of idiopathic hemifacial spasm is described, and experience with its use on 15 patients also described. Experiences in the operative treatment of 6 patients with essential blepharospasm are recounted.


Growth rate and recurrence of residual epidermoid cholesteatoma after tympanoplasty. One hundred and forty-one consecutive patients with chronic otitis media and cholesteatoma treated mainly by open surgical techniques between 1965 and 1972 were studied to investigate the phenomenon of residual cholesteatoma. The average rate of emergence of residual cholesteatoma was 0.25% of the cases per month during the first 5 years, after which the rate of discovery almost ceased. Observed cumulative frequencies of recurrence were 1.4% at 10 months, 14.9% at 5 years and 17.0% at 8.3 years postoperatively. The null hypothesis of no apparent predilection of cholesteatoma for recurrence at any one site was retained. Parameters of growth rates of epidermoid cysts arising from residual cholesteatoma were estimated. Epitympanic cysts grew fairly rapidly, doubling in diameter every 10 months. Mastoid cysts were slower, doubling in diameter every 25 months. Results for the mesotympanum were inconclusive. The practical implications of this information for the management of future cases is discussed.


Percutaneous teflon injection of a vocal cord under fibre optic laryngoscopic control. The technique of percutaneous injection of Teflon paste into a paralysed abducted vocal cord using a fibre optic laryngoscope to observe and control the effect is described. Its use in vocal cord paralysis caused by inoperable bronchogenic carcinoma and the results in 14 patients are discussed.


Transtympanic electrocochleography in the diagnosis of retrocochlear tumours. Transtympanic electrocochleography has been used as a diagnostic procedure in over 250 adult patients with sensorineural deafness. No complications have been encountered. Among these patients vestibulocochlear Schwannoma (acoustic neuroma) has been confirmed in 56 ears while 11 other ears have been found to be affected by other space-occupying lesions. As a result of our observations in these patients, we would like to propose that there are at least three separate criteria to be considered in reaching or strongly suspecting a diagnosis of such pathology. These are broadening of the action potential (loss of P1), observation of a clear microphonic response, and preservation of the action potential even when using stimulus intensities which are not audible in the patients' affected ears. The merits of each of these criteria are discussed and compared with other clinical and operative findings. With the electrocochleographic techniques now employed in searching for acoustic neuromas, false negative results are rare though false positive results are still obtained.


Mastoid surgery: the swing of the pendulum.


Salivary gland chromosomes of Lucilia cuprina (Diptera, Caliphoridae). The salivary gland chromosome complement in Lucilia cuprina consists of five pairs of autosomes and two sex chromosomes X and Y, all with characteristic banding patterns. A standard map for the salivary gland chromosomes of this species has been prepared, and compared with the known map of Lucilia cuprina dorsalis from Australia. The differences in their banding patterns indicate the existence of different varieties within the same species.


A second method for induction of a transmissible entity in the control of cancer in mice. Spontaneous tumour-bearing mice of the C3H/St and subline C3HB/St inbred strains received injections of three nucleosides, adenosine, 6-methyl adenosine, and 5-methyl cytidine, prepared in the same molecular concentration as they had occurred in an alcohol-soluble liver extract which in previous work had suppressed tumour growth. Two transmissible entities appear to be present and they show different degrees of effect upon the growth and regression of spontaneous tumours of mammary gland origin (adenocarcinoma). The first transmissible entity (TE) occurred in the lineal descent of mice following injection of the liver extract. The second entity (TE2) appeared following injection of 5-methyl cytidine. TE manifested its maximum effect in suppressing cancer in mice during the 20th generation of a lineal descent following the injection of the original liver extract into a mother of the cancer proband. TE2 appears to be optimal in controlling cancer in the F11 generation of a second lineal descent derived from an original cancer proband of the C3H/St inbred strain injected with 5-methyl cytidine.


Is extrasensory perception extrasensory?


Ultrastructural and physiological changes induced by atractylate in the canine myocardium. The myocardial cellular response to the cardiac glycoside atractylate, a known inhibitor of mitochondrial adenine nucleotide translocation, was examined physiologically, morphologically, and morphometrically in eight dogs. An isolated in situ segment of left ventricular tissue was employed in these experiments to eliminate collateral flow and the potential for large sampling error. It was found that atractylate infusion rapidly induced physiological and morphological changes suggestive of a loss of membrane regulation of ion flux in the arteriole, capillary endothelium, and myocardial cell. Also mitochondrial changes suggestive of adenine nucleotide translocase inhibition were observed. As these responses are similar to those induced by acute myocardial ischaemia, it is suggested that ischaemia and atractylate induce cell injury by similar mechanisms.


Clinical drug information profit or knowledge orientated.


The use of fibroblast and epidermal growth factors to lower the serum requirement for growth of normal diploid cells in early passage: a new method for cloning. The proliferation of animal cells in tissue culture requires unidentified macromolecules that are contained in serum. Since serum is an extremely complex mixture, efforts to purify these growth-promoting components of serum have met only limited success. On the assumption that these growth-promoting macromolecules in serum are synthesized in active form in some tissue other than blood, we investigated the possibility that they may be purified more readily from specific organs than from serum. We have concentrated our efforts on two likely sources: the pituitary gland, since partially-purified preparations of pituitary hormones have been reported to stimulate the growth of cells in vitro and brain, since neural tissue is essential for the regrowth of amputated limbs in lower vertebrates capable of regeneration. Both pituitary and brain have yielded a strongly mitogenic polypeptide that we call Fibroblast Growth Factor (FGF). FGF was named on the basis of early studies demonstrating its effect on fibroblastic cells: 3T3 cells, amniotic fluid-derived cells, and fibroblasts from human foreskin and mouse embryos. Further work has revealed that FGF promotes the growth of several other mesoderm-derived cell types including adrenal cells, chondrocytes, myoblasts, vascular smooth muscle cells, and vascular endothelial cells. The effect of FGF will be compared to that of EGF and its use for cloning mammalian cells will be described.


Large-scale cultivation of cells in closed systems: stationary and moving. Basic metabolic differences have been observed for cell cultures propagated in conventional stationary or rotating systems. The influence of these differences on virus characteristics is largely unknown but could be significant in maintenance of genetic stability of attenuated strains. Current requirements for good manufacturing procedures prompted the development of closed systems for both stationary and rotating cultures. Both are suitable for large-scale virus production. The methods will be described along with data on yields of a number of vaccine viruses.


Development of a perfusion culture system for production of biologicals using contact dependent cells. Design and evaluation of a new type of culture vessel for contact dependent cells is described. Cells are grown on both sides of stationary glass plates in a laminar flow of medium supplied from separate reservoirs. Construction and operation of vessels ranging in growth surface from 0.02 to 10 m2 is relatively simple and there are no moving parts. Cells can be inspected by low power microscopy in the usual way. Evaluation of this equipment was carried out using the human diploid cell line MRC-5. Saturation densities of up to 1 million cells per square centimetre of growth surface were obtained where appropriate medium volumes were used. Application of this sytem to other cell types and the production of biological products is briefly discussed.


Updated results on the karyology of the WI-38, MRC-5 and MRC-9 cell strains. Data concerning cumulative results obtained from studies of the karyology of the characterized MRC-5 cell strain and the MRC-9 strain at present being characterized, indicate that these cells have karyological characteristics similar to each other and to the extensively studied WI-38 cell strain. The general inference from these studies is that the cells are karyotypically stable and that their karyological characteristics are within the limits recommended for normal cell populations.


Reappraisal of karyological monitoring of diploid cell substrates. The practical aspects of present requirements for karyological monitoring of diploid human fibroblasts used in vaccine production are critically re-examined. A brief review of known chromosomal instabilities and their correlations with neoplasia and with in vitro conversions of cell growth is presented and discussed in relation to karyological criteria in the selection of mammalian diploid cell substrates. It is proposed that extensive experience with the growth and karyological characteristics of human diploid fibroblast cell strains of embryonic origin may warrant the reduction or elimination of karyological monitoring of production lots. Additional requirements for characterization of cell strains for approval as substrates for vaccines are suggested which involve evaluations of chromosomal stability based upon (a) viral transformation rate and (b) aberration production rate in response to x-irradiation.


Karyological characterization of WI-38 cells used in the production of vaccines. According to the valid requirements, samples of WI-38 cells taken prior to the final subcultivation for vaccine production by the manufacturer, were reconstituted, subcultivated and investigated in our control laboratories. Chromosome monitoring was done using permanent orcein stained slide preparations for direct evaluation and photographs of all selected cells for exact counts, scoring for breaks and karyotype analysis in the case of abnormalities. The tabulated summary of karyological analyses on more than 2000 cells shows that with one exception, the chromosome abnormalities did not exceed the upper limits of karyological acceptability. Photographs of abnormalities detected are presented as well as metaphases and reconstructed karyotypes. Our data are compared with those provided by the manufacturer. Obvious discrepancies in the frequency of abnormalities might be due to higher passage levels of the identical cell material tested by us and to slight differences in the methods of preparation and evaluation.


The large-scale cultivation of diploid cell strains in microcarrier culture. Improvement of microcarriers. The use of diploid cell strains for virus vaccine production is mostly retarded by the inefficient culture techniques used for the propagation of these cells. The microcarrier culture system might be a good solution for this problem. DEAE-gels appear to be the most satisfactory microcarrier. The best results are obtained by using DEAE-gels with a rather low anionic exchange capacity or at higher capacity after coating with polymers.


Observations of the incidence of structural abnormalities in the chromosomes of WI-38 cells prepared for vaccine manufacture. Chromosome monitoring of 500 batches of WI-38 cells prepared for vaccine manufacture involved examining 50-60,000 metaphases at p31-32. Results collected over 6 years showed the mean incidence of aneuploidy and breaks to decrease slightly, whereas the incidence of structural abnormalities followed a sharply rising trend, eventually reaching a level where a substantial proportion of cell batches exceeded the limit set for this parameter for vaccine substrates. No technical or cultural factor accounted for the trend which appeared to be correlated with the date of receipt of the seed ampoule. The confidence limits set for this parameter in 1971 were regarded then as provisional. It is suggested that in the light of the observations reported, the position should now be reconsidered.


Production and control of rabies vaccines made on diploid cells. Introductory remarks on the advantages of antirabies vaccines obtained from human diploid cell cultures in comparison with brain tissue vaccine and duck embryo vaccine. The characteristics of these new types of vaccine are: (i) high antigenicity, (ii) rapid development of antibodies, (iii) absence of adverse reactions even when the booster inoculation was given two years later. The disadvantage is the high cost of production to obtain a highly purified product.


The need for non-cultural methods for the detection of mycoplasma contaminants. Mycoplasmas are common contaminants of cell cultures and are of great importance because of the deleterious effects they have upon the infected host cells. Routine detection testing usually relies upon cultural methods and demonstration of characteristic colonies on agar. Questions as to the efficacy of this method have been raised because of increasing realization of the presence of "non-cultivable" mycoplasmas. Several non-cultural detection methods have been developed to aid in the detection of these fastidious microorganisms. These methods include morphology by light and electron microscopy, immunofluorescence, enzyme assays, autoradiography, sucrose gradient separation, altered transport of nucleic acid precursors and altered nucleic acid profiles. A comparison of the relative sensitivities of these methods will be outlined. The need for, and the feasibility of non-cultural detection methods as quality control tests in human virus vaccine production will be discussed.


Clinical responses to T(n)BP-disrupted HDCS (WI-38) rabies vaccine. Study of 10 batches of an antirabies vaccine made from purified, concentrated subunits (Pitman-Moore strain adapted to Wistar HDCS). One single dose of vaccine is insufficient; three or more doses induce high antibody levels. Among more than 2000 vaccines secondary reactions were rare and generally mild and transient. All the individuals vaccinated post-exposure remained without rabic symptoms. The vaccine was also particularly efficacious for booster doses.


Post-exposure use of human diploid cell culture rabies vaccine. 880 individuals, 120 of which were exposed to rabid animals, were immunized pre- or post-exposure with 2 different BPL-inactivated and concentrated rabies vaccines prepared in HDC strains WI-38 and MRC-5. The vaccines were well tolerated and no major side effects were observed after primary immunization with 3-10 doses or 1 booster vaccination. The dynamics of neutralizing, antibody formation and persistence of antibodies in 4 different groups of vaccinees are described. The groups were vaccinated pre-exposure (I) on days 0, 28 and 56; (II) on days 0, 7 and 14; (III) on days 0, 3, 7 and 21; and (IV) post-exposure on days 0, 3, 7, 14, 30 and 90. High antibody levels--persisting for at least 30 months--were obtained in all patients. The CFT, using a concentrated and purified virion antigen, was highly specific for rabies virus antibody demonstration. Since in some 50 patients under severe risk, after having been bitten and/or scratched by proven rabid animals, not a single breakthough of immunity was observed during an observation time between 1/2 and 3 years, the protective effect of the HDCS-rabies vaccines seems to be excellent. With regard to their high immunogenicity and extremely low reactogenicity, the new HDCS-vaccines can be recommended for prophylactic and post-exposure immunization of man without any reserve. Data on simultaneous application of homologous anti-rabies gammaglobulin from man (20 I.E./kg body-weight) and HDCS-vaccines are also presented and discussed.


The nutrient requirements of MRC-5 human diploid cells. Nutrients were assayed by their effect on maximum cell yield (maximum cell population less inoculum) of MRC-5 cells previously maintained in Eagle's basal medium with 10% (v/v) serum, trypsinised, centrifuged and washed. When nonlimiting amounts of iron, methylcellulose and a 68-component supplement were included in our defined medium, cell yields obtained were equivalent to those obtained with 2 to 3% whole serum. Growth then became limited by serum growth factors which, when serum was fractionated by the low temperature ethanol procedure, appeared to precipitate with the alpha-globulins, although the distribution of activity varied from batch to batch of serum. Column chromatography (DEAE and concanavalin A-Sepharose) of serum resulted in much (20 to 45%) loss of growth factor activity during exhaustive dialysis or Sephadex G-25 desalting suggesting that the growth factor may be a diffusible molecule bound in serum to a large protein carrier.


Production of RA27/3 rubella vaccine and clinical results with the vaccine. Adaptation of rubella virus to human diploid cell strains was achieved in 1964, with attenuation in the same cells being accomplished by 1967. The production of rubella vaccine in HDCS is featured by the long period required to build up high titer. High multiplicity of infection gives optimal results. Since rubella virus does not produce a cytopathic effect in HDCS, virus harvests must be made blindly and titrated individually before pooling. A unique feature of this cell-virus relationship is the continuous virus production which takes place for months. For vaccine purposes, however, virus-containing supernatant fluids may be harvested each 48 hrs from the 5th to the 21st day post-infection. Lyophilization presents no particular problems and final titers in ampoules can be as high as 10(5.0) PFU, which represents about 100 subcutaneous doses. Control of rubella vaccine presents only the problem that viral CPE is absent or mild. Therefore the presence or absence of virus may have to be determined by interference. Titration of rubella virus produced in HDCS must be by plaquing in RK13 rabbit kidney cells or by interference in cynomolgus monkey kidney cells. Rubella vaccine (RA27/3 strain) produced in HDCS has been more immunogenic than other strains and retains an ability to infect intranasally as well as subcutaneously.


Mislabeling, contamination, and other sins of cultured flesh.


Towards a chemically defined medium for the growth of normal human diploid cells.


Tumourigenicity testing in immunosuppressed mice: advantages and disadvantages. Optimal conditions for the detection of malignant cells substrates were investigated. Three approaches were used: (1) neonatally thymectomized, antithymocyte serum (ATS) treated mice, (2) thymectomized, lethally irriadiated, bone marrow reconstituted (T-B+mice, and (3) congenitally athymic nu/nu (nude) mice. All three systems successfully distinguished between normal and malignant cell lines. However, differences were observed between different colonies of nude mice as to their capacity to support progressive growth of certain tumours. Advantages and disadvantages of the immunosuppressed mouse model system will be discussed. They will include: (a) detection of malignant cells in mixed populations, (b) need for different routes of inoculation for different cell types, (c) comparison with other malignancy testing systems, (d) long term versus short term immunosuppression.


The significance of positive controls in testing the oncogenic potential of cell material for virus vaccine production. For all the parts involved in the production and control of virus vaccine it is important that controls of the cell material should be carried out under optimal conditions for the detection of any abnormal characteristics of the cells. The experiments reported here attempt to establish some of the optimal parameters for testing out the tumor-inducing properties of a human diploid cell culture. All experiments were carried out on 6-week old Syrian hamsters and the main objectives were: (i) to devise an optimal immunosupression scheme and (ii) to find a more suitable positive control for testing the oncogenic potential of the human diploid cells. The anti-hamster lymphocyte serum proved to be a more potent immunosuppresser than hydrocortisone succinate as seen from the development of tumors following injections with KB cells; this scheme was subsequently used in assessing the usefulness of KB and Swiss-12 cells as positive controls. The experiments lend support to the suggestion that a heteroploid fibroblastic culture (such as Swiss-12) should become a standard positive control since not only are the tumors so obtained larger and less prone to early necrosis but, more important, they bear a closer resemblance to the kind of tumors likely to be induced by the diploid cell cultures being tested.


Use of human diploid cell MRC-5, for production of measles and rubella virus vaccines. A further attenuated strain of measles virus, called AIK strain, developed by Makino and his colleagues at the Kitasato Institute, Tokyo, was adapted to human diploid cells (HDC), MRC-5. In a field trial it was found that this strain initiates a low rate of clinical reactions and 100% seroconversion in vaccines. An attenuated strain of rubella virus, called Takahashi strain, isolated by Shishido at the National Institute of Health, Japan, was also adapted to HDC, MRC-5. Brief details of production of an experimental rubella vaccine are given. Up to 20 harvests of virus were made from each culture without any apparent alteration of diploid cells.


Effects of vagotomy on the catecholamine-containing cells in the gastric mucosa of the rat: an electron microscopic study. The ECL cells in the fundic mucosa from five different groups of fasted rats were examined by means of electron microscopy; in the control animals, the majority of the secretory granule of the ECL cells contained electron-dense material. Previous studies in our laboratory indicate that this material probably represents a catecholamine. In the second and third groups, in which truncal vagotomy plus pyloroplasty was performed, the secretory granules were depleted of their content. In the fourth group in which vagotomized animals were treated with iproniazid and L-dopa, the mature secretory granules were depleted but formation of new granules was apparent. In the fifth group, in which pyloroplasty alone had been performed, the content of the granules was the same as in the control group. It was concluded that the endocrine function of the ECL cells is under vagal control and that vagotomy causes release of catecholamines from these cells.


Micro-organisms in the midgut of tsetse fly larvae. Two types of micro-organisms were found in the midgut of Glossina morsitans larvae, a large Gram-negative bacterial rod and a small Gram-negative rickettsia-like micro-organism, although the occurrence of the rickettsial type is restricted. The location of these micro-organisms in a small area of the proventriculus of all three larval instars is discussed. The large micro-organisms resemble milk-gland bacteria, and further evidence is presented in support of a milk transmission hypothesis for these micro-organisms.


Studies on the bacteriophage PS8 of Agrobacterium tumefaciens (Smith and Townsend) Conn: physico-chemical properties of its DNA. DNA from the bacteriophage PS8 was extracted and purified. The buoyant density was determined was 1.716 cm3/g. The guanine-cytosine content was calculated to be 57%. DNA molecules which looked like circles were found among linear strands in an electron-microscopic study. With an endonuclease from Streptomyces albus G the DNA was digested to 19 fragments, with molecular weights ranging from 600 to 7,400 daltons. The molecular weight of the DNA was determined to be 38.8 X 10(6) daltons +/- 8.7%.


Radiological diagnosis of posterior fossa tumours in childhood.


The vascular malformations of the vertebrobasilar trunk in infancy.


Various developmental anomalies of the posterior fossa. A radio-pathological study.


Magnification angiography of the posterior cranial fossa in pediatry.


The arteries of the posterior cranial fossa in new-born.


Problems of diagnosis and treatment in the Dandy-Walker syndrome.


Congenital atresia of the foramen of Magendie, so-called Dandy-Walker syndrome.


Late death in cases of treated internal hydrocephalus caused by encephalo-cranial disproportion.


Surgical management of Chiari malformation in children.


Malformation of the cranio-vertebral region. Technical notes and case report.


Malformative pathology of the medial structures of the posterior fossa: considerations on 17 cases.


Surgical treatment of encephalocele complicated by hydrocephalus.


Cystic malformation of the aqueduct and of the fourth ventricle.


Hypoplasia of the tentorium with pseudohernia of the occipital lobe into the posterior cranial fossa.


Echographic and biochemical methods for evaluating hydrocephalus and shunting procedures.


CT (computerized tomography) in paediatric neurosurgery. Results of a survey of 400 patients.


The treatment of posterior fossa tumours in childhood.


Membranoproliferative glomerulonephritis associated with infected ventriculoatrial shunt. Report of two cases recovered after removal of the shunt.


A new technical method of ventriculoatrial shunt implantation.


The medical treatment of hydrocephalus using isosorbide.


Reconstitution of the cerebral mantle in shunted hydrocephalic children.


The possible causes of good long-term results following surgical treatment of hydrocephalus.


Follow-up study of 360 children subjected to extracranial CSF shunt for hydrocephalus.


Management and prognosis of gliomas of the optic pathways in children. A follow-up study was made of 24 children with glioma of the optic pathways. In gliomas restricted to one optic nerve, total excision should be performed. The prognosis is excellent. Radiotherapy is not indicated. In gliomas of the anterior chiasm, a biopsy should be taken; occasionally, partial removal is indicated. The prognosis is good. Radiotherapy is indicated only if the follow-up shows a progression in the visual signs. In most cases, the growth of the tumor seems to be arrested. The vision remains stable and most patients have a useful degree of vision in at least one eye. In gliomas of the posterior chiasm with hypothalamic signs, the prognosis is poor. A biopsy and radiotherapy are indicated.


Cerebrospinal fluid absorption studies in patients with myelomeningocele and hydrocephalus. Preliminary report.


Results of microsurgical treatment of gliomas of the optic system and the hypothalamus in children.


Operative indications in dysraphic malformations. Follow-up studies and late results.


Pituitary adenomas in infancy and childhood.


Unusual features of an orbital neurofibroma.


The present diagnostic approach of the mass lesions of the posterior fossa in children.


Neuroradiological and longitudinal studies of the pineal region tumors.


A method in surgical treatment of premature closure of metopic suture (trigonocephaly).


Pathogenetic hypothesis of intracortical calcifications in Sturge-Weber disease. Case report following lobectomy.


Section of the portio minor of the trigeminal nerve for the treatment of the Marcus Gunn phenomenon.


Brain abscess in children.

Brain abscess in children.


Brain tumor with ipsilateral cerebral hemiatrophy in children.


Kleeblattschädel syndrome. Report of a treated case.


Hydatid cyst of the fourth ventricle in a six-year-old child.


Features of the clinical evolution following the acute stage in severe traumatic coma in childhood. A study of 45 cases of so-called apallic syndrome.


Severe head injury without mass lesion in children. The role of intracranial hypertension.


Cerebrospinal fluid derivation in posterior cranial fossa tumor with hydrocephalus.


Morbid anatomy of the tumours of the posterior fossa in childhood.


Ventricular fluid pressure in posterior fossa tumors of childhood.


Management of hydrocephalus secondary to posterior fossa tumors.


Gas study in arrested hydrocephalus with chiasmatic block.


The value of preoperative shunts in posterior fossa tumors.


Clinical and diagnostic problems in posterior fossa tumors in children (analysis of 100 cases).


Posterior fossa tumors in childhood.

Posterior fossa tumors in childhood.


New problems in the therapy of medulloblastomas.


Postoperative hyperthermia following posterior fossa craniotomy.


Tumoral versus non-tumoral posterior fossa lesions. A clinical diagnostic problem.


Operative problems in encephalocele.

Operative problems in encephalocele.


Experiences with radiotherapy and chemotherapy in the treatment of tumors involving the caudal brain stem.


Treatment and prevention of complications following surgery for tumors involving the posterior cranial fossa.


Primary spinal cord tumors in infancy and childhood. Remarks on 53 operated cases.


Posterior fossa tuberculomas.

Posterior fossa tuberculomas.


Mucins from cystic fibrosis sputum.

Mucins from cystic fibrosis sputum.


The value of computerized axial tomography (CAT) in early diagnosis of intracerebellar hemorrhage in children.


Diagnosis of posterior fossa malformations in infants and children using axial computerized tomography (CT).


Mucous glycoproteins.

Mucous glycoproteins.


Tracheobronchial mucus secretion in vivo and in vitro by epithelial tissues from cystic fibrosis and control subjects.


The composition of human gastric mucin.

The composition of human gastric mucin.


Synthesis and secretion of respiratory tract mucins by tracheal explants.


Bronchial mucus secretion in cystic fibrosis.


Pathophysiology of mucus secretion in cystic fibrosis.


Season of birth and Schneider-oriented diagnosis of affective disorder. Most earlier studies and all recent studies on national samples have shown that compared with live births in the control population, schizophrenic patients have a significant excess of birth rates in the winter or early months of the year. In contrast, only some of the early research efforts and only some of the national studies (in England and Wales as well as for certain decades in Sweden) have demonstrated that the same holds true for patients with affective psychosis. The present German study, carried out on affective disorder diagnosed in a strongly Kurt Schneider-oriented clinic, found (as did most Scandinavian research on national samples) that there was no significant overrepresentation of births in the winter or early months of the year for all types of affective disorder, neurotic as well as psychotic. Thus, the findings on Schneider-diagnosed affective disturbances were similar to those on Schneider-diagnosed schizophrenia reported elsewhere.


Death anxiety in normal children and adolescents. Instruments devised to measure death anxiety, depression, and manifest anxiety in adults were administered to 75 junior high-school students, 111 senior high-school students, and 38 adults in an effort ot determine the nature of any developmental differences among these groups. A number of significant differences were noted, with the senior high-school students showing greater anxiety, depression, and death anxiety than the other groups. These results are discussed in the context of tasks of adolescent development, and an item analysis of the death anziety questionnaire is presented. Developmental differences in the origin of death anxiety are related to previous studies of death anxiety in adults.


Rheological factors in mucociliary clearance. The assessment of mucotropic agents using an in vitro model.


Mucus rheology and mucociliary clearance.


Acoustic neurinomas in infancy. Report on four cases.


Impairment of mucociliary transport in cystic fibrosis.


Mucociliary clearance in cystic fibrosis.


Studies of mucus permeability. Water transport through intestinal mucus layers.


Synthesis and intracellular transport of glycoproteins.


Goblet cells and secretion of mucus.

Goblet cells and secretion of mucus.


The effect of mucus cross-linking on the clearance of mucus.


Some physical properties of mucins and their molecular origin.


Morphological and biochemical approaches to characterize the structure and function of the golgi complex.


Isometric dynamic response of mammalian heart muscle due to step changes in the calcium concentration of the perfusing medium. The isometric tension transient response of isolated kitten papillary muscle was found to exhibit direction-dependent dynamics when subjected to step changes in the calcium concentration of the bathing medium. The tension transient curves could be resolved into a minimum of two to three exponential components, which could be considered to indicate different calcium compounds or compartments within the muscle. However, a model relating calcium diffusion across the muscle to the dose-response curve indicates that the exponential components of the tension transient curves are due to the properties of calcium diffusion through the extracellular space, and that the direction-dependent dynamics are caused by the nonlinearity of the dose-response curve.


Possible mechanism of rate-dependent change of contraction in dog ventricular muscle: relation to calcium movements.


Glycosyltransferases and hypersecretion of respiratory mucus.


Dynamics of contraction with special reference to calcium. Using a new method for tetanizing frog ventricle muscle, the tension-length, force-load-velocity, and load-extension relations were determined under the steady active state. On the basis of these experiments a mechanochemical model was developed. Time courses of internal calcium concentrations, the rates of release, and uptake of calcium during isometric twitches were calculated by the model.


Delay in diastolic restitution of full myocardial contractility in guinea pig atrium. In an isolated perfused left atrium of guinea pig, pressure amplitudes were recorded at 37 degrees C and a stimulation rate of 240/min with interposed variable resting pauses. After prolonged intervals the preparation develops maximum pressure, which does not depend on perfusion time or on any positive inotropic intervention. The latter is found to shorten the time needed for restitution of full contractility. Nifedipine (5 X 10(-7)M) exerts its negative inotropic effect mainly by a delay in restitution time; acetylcholine (2 X 10(-7)M), by a concomitant reduction of all pressure amplitudes over the whole range of stimulus intervals. The latter effect is similar to that of lowering [Ca2+]o.


Contraction duration and diastolic stiffness in aged canine left ventricle. The data obtained from groups of young and old beagles demonstrate that senescent beagle hearts are characterized by lower contractile function as measured by developed pressure and maximal rate of pressure change with time, by longer contraction duration, and by increased dynamic stiffness per given pressure as measured by the sinusoidal forcing technique.


Voltage-clamp experiments by double microelectrode technique in rabbit sinoatrial node cell. When a man-made strand of the rabbit sinoatrial node (S-A node) was shortened by ligation, the spatial decay of the electrotonic potential decreased and the input impedance increased. A satisfactory voltage clamp was achieved in the small S-A node specimen using the double microelectrode technique (Deck, Kern, and Trautwein, 1964).


Interaction of lanthanides with muscle microsomes. The interaction of gadolinium, a lanthanide with calcium-blocking action, with isolated muscle microsomes has been studied. Two classes of binding sites were present, 80 and in excess of 300 nmol/mg in number, respectively. Divalent cation, including Ca, Zn, and Cd, blocked the higher affinity site. Antibiotic ionophores X537A and A23187 enhanced binding, with positive cooperativity. ATP enhanced binding at low concentration (10-20 muM) of nucleotide, and without ATP-hydrolysis. The data suggest a dissociation in the intact membrane between the binding and ATP-hydrolytic portion of the transport site.


Subcellular localization of cardiac adenylate cyclase: sarcolemma or sarcoplasmic reticulum?


The relation of structure to function in gastric mucus.


Role of slow inward current on premature excitation in ventricular muscle. Action potential durations in premature excitations showed paradoxical prolongation at the shorter coupling intervals; this was abolished by manganous ions. Voltage-clamp experiments also disclosed a transient increase of slow inward current in premature excitations. These results indicate that prolongation of action potential durations was mainly brought about by changes in slow inward current, especially in its characteristics of recovery from inactivation.


A new spectroscopic approach to cardiac energy metabolism. The optical characteristics of the hemoglobin-free perfused rat heart have been examined in detail. For dual-wavelength spectrophotometry, absorption pairs at 605-620 nm and 587-620 nm are found to be suited for the investigation of the oxidation-reduction changes of cytochrome aa3 and oxygenation-deoxygenation of myoglobin in cardiac tissue. Techniques by which the absorption changes of myoglobin and cytochrome aa3 can be measured during one cycle of contraction-relaxation are presented. The results demonstrate that, in the aerobic state, myoglobin is more oxygenated during the systolic and diastolic periods and deoxygenated in the resting period, whereas cytochrome aa3 is more reduced in the systole and diastole and oxidized in the resting period.


Significance of heat loss in energetics of left ventricle. Left ventricular heat loss was examined by measuring the temperatures in the coronary sinus blood, in the aortic blood, and in the left atrial blood by thermistors. The temperature was highest in the coronary sinus blood, in the aortic blood next, and it was the least in the left atrial blood. However, since the aortic blood flow was naturally higher than the coronary sinus blood flow, the heat loss was larger in the left ventricular cavity than in the rest of the chambers. This heat loss was mostly proportional to the magnitude of the changes of both the arterial blood pressure and the aortic blood flow.


Metabolism of individual fatty acids in heart muscle. We have studied the incorporation of various long chain fatty acids into tissue lipids using various heart preparations. We found that individual fatty acids are incorporated into various lipid fractions at unequal rates. In particular, erucic acid was shown to be a poor acyl donor for various acylation reactions. Labeling of triacylglycerol by exogenous fatty acid was found to be extremely rapid in the perfused rat heart and in the in vivo heart.


Uptake and release of immunoreactive insulin in coronary circulation in man: studies at rest, during exercise, and during glucose and insulin infusions. Significant uptake and release of immunoreactive insulin by the heart have been observed in man, and this is related to plasma insulin levels. Exercise and the fed state appear to affect the myocardial handling of insulin. The findings could not be related to myocardial carbohydrate metabolism, but could, during exercise, be related to myocardial lipid metabolism.


Transmural mitochondrial differences in myocardium. Two functional mitochondrial populations with different sedimentation rates (S) were obtained from homogenates of canine myocardium by rate zonal centrifugation using an iso-osmotic Ficoll gradient. To ascertain the origin of these populations, the left ventricular wall of normal myocardium was divided into subepicardial (outer one-third), intermediate (middle on-third), and subendocardial (inner one-third) layers. The slow S mitochondria comprised 75% of the mitochondrial population of the subepicardial layer. In contrast, the fast S mitochondria contributed 65% of the subendocardial population. Intermediate layer mitochondria resembled those of the subepicardium. Mitochondria isolated from the three layers had approximately the same density, as shown by isopycnic zonal centrifugation. These studies indicate that mitochondria from subepicardial and subendocardial layers of normal myocardium differ in size and shape but not in density. Electron micrographs (EM) of the subepicardium showed many mitochondria as long as 4 to 8 sarcomeres. Mitochondria from the outer and inner layers of normal myocardium had the same oxidative phosphorylation parameters. Acute myocardial infarction, lasting 1 or 2 hr, resulted in the selective loss of the fast S mitochondria. Because the fast S mitochondria are prevalent in the subendocardium, these results may explain the greater vulnerability of this layer to anoxia.


Myocardial adenine nucleotides after infusion of adenosine. In the rabbit heart, continuous infusion (12 ml/hr) of 1% adenosine into the superior caval vein, or of 0.5% adenosine into the left myocardial ventricle, for a period of 3 hr increased the myocardial tissue level of ATP by almost 40% over the control (5.0 mumol/g). Both shorter or longer periods of application and lower or higher concentrations of adenosine, as well as the infusion of adenosine into the left atrium, were less effective. The increase markedly outlasted the period of application of adenosine. It did not result from the adenosine-induced systemic hypotension and decrease in cardiac work, but must be attributed to a direct metabolic effect of the adenosine infusion.


Myocardial fatty acid and cardiac performance. Isoproterenol enhanced myocardial triglyceride utilization and incorporation of exogenous fatty acid into triglyceride fraction in the perfused rat heart in the presence of glucose. However, in the absence of glucose, cardiac performance was considerably aggravated by isoproterenol. This suggests that glucose might be an essential fuel in the heart stimulated by isoproterenol.


Influence of carbocromene on free fatty acid metabolism of the heart. In view of the fact that carbocromene (C) influences the metabolic processes in the heart muscle, its effect on the transport of substrates into the heart tissue was investigated. In rat heart slices C inhibits the uptake of [1-14C]palmitic acid up to 40%, while 14CO2 production is reduced to the same extent. At the same time oxygen consumption diminished by about 20%. Using 3-O-[14C] methyl-D-glucose as metabolic marker it was shown that the uptake of glucose into the heart tissue increased by 30% under the influence of C. From these results it can be concluded that C inhibits the oxidation of FFA in favor of glucose and diminishes the oxygen-wasting effect of FFA.


Electrically induced automaticity in canine ventricular myocardium. Repetitive spontaneous action potentials (SAP) could be induced in canine ventricular and atrial muscle, although this inhibitory action was antagonized by the pretreatment with voltage range between about -60 mV and 0 mV. The SAP seemed dependent on both slow inward Ca2+ and Na+ currents and was suppressed by verapamil, Mn2+, and diltiazem, but not by tetrodotoxin. The increase of extracellular potassium concentration also suppressed the SAP. Acetylcholine could not block the SAP in ventricular muscle, but inhibited that in atrial muscle, although this inhibitory action was antagonized by the pretreatment with atropine. The automatic activity was attributed to slow inward Ca2+ and Na+ currents modified by decreasing time-dependent K+ outward current and K+ anomalous rectification.


Effects of insulin on mammalian cardiac muscle. Insulin increases the contractile force of myocardium with hyperpolarization and a faster rate of fall of the action-potential plateau. This phenomenon is augmented inthe reduced-K+ or ouabain solution. It is suggested that the insulin-induced positive inotropism can be explained by facilitation of inward movement of Ca2+, which possibly is associated with an increase in outward movement of K+.


Free fatty acid content of myocardial interstitial spaces of dog. Morphological and biochemical observations from our laboratory have shown the presence of lipids in the cardiac interstitial spaces of the dog. The present study was designed to assess the importance of free fatty acids (FFA) in these lipids using FFA or sucrose tracers in 14 anesthetized fasting open-chest mongrel dogs. Endogenous FFA and labeled tracers were measured in arterial and coronary sinus plasma; they were also determined in lymph collected from cardiac efferent lymphatic trunks. [14C] sucrose was infused at a constant rate in the femoral vein of 5 dogs. The concentration of the tracer in the lymph was 90% of the arterial concentration after 60 min of infusion. On the other hand, when [1-14C] palmitate was infused at the same rate in 7 dogs, the ratio of lymph to arterial tracer concentration was only 20% (60 min) and 25% (120 min), even though the myocardial extraction and oxidation of the tracer were stable. This ratio tended to reach a plateau (greater than or equal to 90%) only 24 hr after a bolus injection of the tracer. This tracer study shows the presence of a pool of myocardial fatty acids with a relatively slow turnover rate that may constitute an important reservoir of FFA.


An experimental study of the effect of glucose-insulin-potassium solution on energy metabolism of infarcted cardiac muscle. The effect of GIK (glucose-insulin-potassium) infusion on the energy liberation of the infarcted heart muscle was studied experimentally in relation to epicardial electrocardiographic changes and cardiac dynamics. In the case of 30-min coronary ligation, the spread of the zone showing ST segment elevation was not depressed significantly, but the mean voltage values of ST elevations were suppressed in the peri-infarcted area, and the uncoupling of oxidative phosphorylation in the peri-infarcted area was also obscured by the GIK treatment. Cardiac dynamics did not change with GIK treatment. On the other hand, in the 60-min coronary ligation, the spread of the zone showing ST segment elevation was reduced significantly and the negative inotropism induced by coronary ligation was suppressed with GIK infusion. However, the uncoupling of oxidative phosphorylation in the infarcted area and in the peri-infarcted area did not show any significant improvement. Therefore, the disturbed energy metabolism of the peri-infarcted area of 30-min ligation might be improved by the effect of GIK infusion. These data suggested that the GIK infusion improved the energy liberation of the so-called twilight zone at 30 min after ligation, and then reduced the genuine infarcted zone after 60 min of ligation.


Calcium paradox: changes in high-energy phosphate compounds of isolated perfused rat hearts. When isolated rat hearts are perfused with Ca2+-containing medium, after a brief Ca2+-free period, irreversible cell damage occurs (calcium paradox). This phenomenon is concomitant with a rapid consumption of myocardial high-energy phosphate stores, prior to the appearance of these compounds in the effluent perfusion medium. A possible mechanism for the origin of myocardial necrosis, caused by intracellular Ca2+ overload, is discussed.


Effect of diltiazem on calcium- and noradrenaline-induced contractions in isolated rabbit aorta. The effect of diltiazem on calcium- and noradrenaline-induced contractions was investigated in the K-depolarized aortic strip of the rabbit. Diltiazem inhibited the contraction induced by calcium ions dose-dependently. A lower concentration of diltiazem produced no significant influence on the contraction induced by noradrenaline, while a higher concentration suppressed it. It is assumed that in the rabbit aorta diltiazem effectively inhibits the contraction mediated by the influx of calcium ions but it is less effective with the contraction induced by the release of sequestered calcium.


Effects of morphine and halothane on canine cardiac function before and after cardiopulmonary bypass. In an effort to compare the effects of anesthetics on cardiac functions and metabolism, mongrel dogs were anesthetized with morphine (3 mg/kg, N2O 50% in oxygen) or halothane (0.7%, N2O 50% in oxygen). Hb, Ht, pO2, pCO2, pH, O2 content, lactate, and pyruvate in the arterial and coronary sinus blood were measured and ECG, LVP, CVP, AP, and CO were recorded. Aorta was cross-clamped for 30 min under cardiopulmonary bypass (CPB). Hemodynamic and metabolic survey were continued until 120 min of recovery period. Results were as follows: Hemodynamics, such as LVP, AP, and CO, were depressed after CPB, then gradually returned to control level. Subendocardial ischemia observed through ST depression and QRS widening on ECG, as well as DPTI/TTI ratio under 1.0, disappeared in the recovery period. Lactate was produced by the heart after anoxic arrest and was extracted at 120 min after CPB. Halothane and morphine did not show any significant differences of effects on cardiac function and metabolism in this study.


Effects of hypoxia and digitalis glycosides on myocardial stiffness. The influences of hypoxia and of the interactions of hypoxia with digoxin and ouabain on myocardial stiffness were studied at two extracellular calcium concentrations (2.5 mM and 4.0 mM) in isolated, isometrically contracting cat papillary muscles. Stiffness (delta T/delta L), defined as a change in tension (delta T) in response to an imposed length change (delta L), was measured during contraction and during rest by use of a sinusoidal forcing function. Neither digoxin, ouabain, nor increased extracellular calcium altered contraction or resting stiffness in the well-oxygenated environment. Resting stiffness was increased at the end of hypoxia only in the presence of digoxin in both 2.5 mM and 4.0 mM Ca. Contraction stiffness was increased in 2.5 mM Ca by hypoxia alone and by hypoxia in the presence of digoxin or ouabain, but was not increased in experiments carried out in 4.0 mM Ca. Thus it appears that hypoxia per se increases contraction stiffness, but increasing extracellular calcium from 2.5 mM to 4.0 mM prevents the increase elicited by hypoxia; resting stiffness, however, is increased by hypoxia only in the presence of digoxin, and this occurs in both 2.5 mM and 4.0 mM Ca.


Inotropic and chronotropic effects of antiarrhythmic agents onisolated blood-perfused canine ventricular tissue. In the isolated blood-perfused canine ventricular tissue, antiarrhythmic agents could be classified as having: 1) positive chronotropic and inotropic effects (procainamide), 2) negative chronotropic but positive inotropic effects (quinidine), and 3) negative chronotropic and inotropic effects (lidocaine, ajmaline, diphenylhydantoin, and propranolol).


Influence of quinidine on ATP-linked calcium binding by heart mitochondria and microsomes. The action of quinidine on heart microsomal and mitochondrial calcium binding in the presence of MgATP was studied under different experimental conditions and compared with other antiarrhythmic agents such as procaine amide and lidocaine. Quinidine stimulated microsomal calcium binding but depressed mitochondrial calcium binding. Although procaine amide stimulated microsomal calcium binding, it did not affect mitochondrial calcium binding. On the other hand, lidocaine depressed calcium binding by mitochondria without affecting calcium binding by the microsomal fraction. The stimulation of microsomal calcium binding by quinidine was not apparent at high concentrations of Mg2+, low concentrations of ATP, or low concentrations of Ca2+. The depressant action of quinidine on mitochondrial calcium binding was not observed at low concentrations of Mg2+ or ATP but was more pronounced at low concentrations of Ca2+. These results suggest that the action of quinidine on mitochondria may play a major role in eliciting cardiodepressant effect.


Effects of halothane anesthesia on myocardial contractility, coronary as well as systemic hemodynamics, and myocardial metabolism in dogs. Halothane depressed coronary and other systemic circulations as arterial halothane concentration rose in dogs. Myocardial oxygen consumption decreased as the cardiac action, especially "myocardial contractility", was depressed, while the whole-body oxygen usage remained steady. Lactate and pyruvate levels in arterial, mixed venous, and coronary venous blood did not show any signs of tissue hypoxia or development of anaerobic metabolism even in very deep halothane anesthesia. Coronary blood flow seemed well maintained for the demand, and changed almost in parallel with the myocardial oxygen consumption during the observation.


Glycolytic ATP and its production during ischemia in isolated Langendorff-perfused rat hearts. The perfused rat heart was used to assess the possible contribution of glycolytically produced ATP to the maintenance of the action potential in the normoxic heart, and to the maintenance of membrane integrity in the underperfused, ischemic heart. During normoxia, pyruvate (10 mM) was nearly as able as glucose (10 mM) to maintain the normal action potential. During ischemia (reduction of perfusion pressure of Langerdorff heart from 100 to 20 cm H2O), total tissue values of ATP and creatine phosphate were similar in pyruvate and in glucose hearts. However, pyruvate-perfused hearts had higher tissue levels of cyclic AMP during the ischemic period, and during the reperfusion period they had an increased release of lactate dehydrogenase and an increased incidence of arrhythmias when compared with glucose hearts. It is proposed that these differences can be related to a higher rate of production of glycolytic ATP. The anatomical, biochemical, and pharmacological evidence favoring a cytoplasmic compartment of ATP located in relation to the cell membrane is reviewed.


Alteration in calcium metabolism in mitochondria isolated from ischemic and reperfused myocardium. This study determines the effect of ischemia and reperfusion on energy-linked Ca2+ uptake by myocardial mitochondria. The left anterior descending coronary artery was occluded in 14 mature pigs for 2 hr. In seven animals the ligature was released and the ischemic zone reperfused for 2 additional hours. After sacrifice, mitochondrial function was measured in normal and reperfused or ischemic areas of the left ventricle, using a polarographic method. Mitochondria were prepared without EDTA by standard procedures and Ca2+ uptake measured by 45Ca2+ isotope tracer. Uptake of Ca2+ by mitochondria derived from ischemic myocardium is markedly impaired with or without phosphate. Reperfusion may accentuate this impairment. The presence of exogenous Ca2+ inhibits the ability of ischemic or reperfused mitochondria to phosphorylate ADP.


Chronotropic effects of calcium and magnesium ions at different temperatures. In the isolated right atrium of rabbit hearts, excess calcium causes an increase of heart rate at 38 degrees C; this effect is absent or even reversed at a temperature of 30 degrees C. From microelectrode studies it seems that when the calcium concentration is increased the pacemaker shifts within the sinoatrial node to cells in which excess calcium causes an acceleration of diastolic depolarization (type A). In fibers where the impulse originates at low calcium (type B), excess calcium causes a deceleration of diastolic depolarization. At low temperature, the effects on type A fibers are completely absent. Excess magnesium has a negative chronotropic effect at both temperatures, mainly by a deceleration of diastolic depolarization in type B fibers.


Influence of ATP or oxygen plus substrate on occurrence of the calcium paradox. Reperfusion of Ca2+-deprived rat hearts with Ca2+-containing medium results in irreversible cell damage (calcium paradox). In this study this type of cell damage was studied in the anoxic rat heart, in the presence and absence of glucose. Creatine kinase (CK) release was used to define cell damage. Hearts were perfused successively with Ca2+-containing medium (30 min), Ca2+-free medium (5 min), and Ca2+-containing medium (5 min). In the presence of glucose, myocardial ATP was maintained at a fairly high concentration. Reperfusion with Ca2+ resulted in an immediate and massive release of CK. In the absence of glucose, the ATP concentration was almost zero after 30 min. Reperfusion with Ca2+ did not result in release of CK. Massive release occurred as soon as these hearts were reoxygenated. It is concluded that this type of calcium-induced cell damage only occurs in the presence of ATP, or oxygen plus substrate. Mitochondria most likely play a major role in the occurrence of the calcium paradox because of their ability to accumulate huge amounts of Ca2+ under these conditions.


Effect of xanthine oxidase inhibitor on myocardial ischemia. The xanthine oxidase inhibitor, 4-hydroxypyrazolo(3,4-d) pyrimidine (HPP), Allopurinol, caused augmentation of myocardial uptake of [3H] hypoxanthine, which was eventually completely incorporated into ATP. The decrease of [32P] orthophosphate incorporation into ATP induced by isoproterenol was restored by HPP administration.


Relationship between energy liberation and utilization in ischemic cardiac muscle. Mitochondrial respiration, succinate dehydrogenase coenzyme Q reductase, and myosin B were investigated in ischemic myocardium from experimental myocardial infarction in dogs. Respiratory control ratio of mitochondria was impaired by ischemia at 60 min after coronary ligation, and oxygen consumption was inhibited 120 min later. Enzyme activity of succinate dehydrogenase coenzyme Q reductase was decreased at 6 hr after coronary ligation. Calcium ion sensitivity of myosin B declined 12 hr after coronary ligation. However, adenosine triphosphatase activity of myosin A from infarcted myocardium was not different from that of the intact one. These results suggest that interaction in the sequence of enzyme complexes was first impaired in ischemic myocardium and that deterioration of enzyme activity was then manifested.


Role of myocardial lipids in development of cardiac necrosis. Significant alterations in fatty acyl composition of cardiac phospholipids and neutral lipids are induced by dietary cod liver oil in the rat. Increased dietary availability of docosahexaenoic acid (22:6 omega3) leads to extensive replacement of linoleic acid (18:2omega6) and arachidonic acid (20:4omega6) in phospholipids. Dietary cod liver oil (10%) reduces isoproterenol stress tolerance and results in increased development of cardiac necrosis and mortality following isoproterenol treatment. It is suggested that diminished catecholamine stress tolerance may be related to altered synthesis of prostaglandins or related products.


Energy liberation in abnormal cardiac muscle. Uncoupling of oxidative phosphorylation was suggested in experiments using the infarcted myocardium, even in dogs with 15-min ligation. The same type of disturbance in mitochondrial respiration was observed in dogs with reperfusion after 45-min ligation, whereas the disturbance was recovered in dogs with reperfusion after 15-min ligation.


Myocardial energy and electrolyte metabolism during exercise in patients with circulatory disease.


Contribution of an electrogenic sodium pump to the response of sinoatrial node cells to acetylcholine. ACh (10-7) - 10(-6) g/ml) induced hyperpolarization in pacemaker cells in the sinoatrial (SA) node of the rabbit heart varied inversely with the level of the maximal diastolic potential of the pacemaker cells. The hyperpolarization induced by ACh was completely abolished at 4 degrees -6 degrees C. Ouabain (10(-6) g/ml) markedly depressed or abolished the hyperpolarizing response. The response was also depressed in K-free solution, but not abolished. In Ca-free or low-Na solution, the sino atrial node cell responded well to ACh, showing marked hyperpolarization. Present results indicate that the hyperpolarization induced by ACh is at least in part due to an electrogenic Na pump activated by ACh.


Disturbance of myocardial energy liberation in experimental charcoal embolism of canine pulmonary artery. Experimental charcoal embolism of the pulmonary artery was produced in dogs. Changes in hemodynamics, electrophysiology, and cardiac energy liberation were observed. The effects of histaminic and antihistaminic drugs on these changes were also studied. After two infusions of charcoal suspension, acute right ventricular overloading was recognized hemodynamically and electrophysiologically. Mitochondria isolated from both ventricles exhibited uncoupling of oxidative phosphorylation polarographically. These changes were enhanced with histamine injection and reduced with diphenhydramine injection. Histamine was seen to have some uncoupling effects on the heart mitochondria and a slight effect on cardiac dynamics. It is concluded that oxidative phosphorylation of the heart mitochondria was uncoupled in the acute stage of experimental pulmonary embolism, and that histamine might emphasize this change directly or indirectly or both, that is, directly affecting the myocardial cell metabolism, and indirectly aggravating myocardial hypoxia through pulmonary vasoconstrictive action.


Intraventricular conduction disturbance due to delayed recovery from ventricular inactivation in chlorpromazine-treated dogs. In in situ canine hearts, chlorpromazine induced a time (preceding cycle length)-dependent decrease in conduction velocity within the ventricle. Thus, QRS duration of nonpremature beats was lengthened at rapid pacing rates while QRS duration of atrial premature beats was lengthened at short coupling intervals. These slow conductions were not due to reduced take-off potential of ventricular action potentials but to drug-induced slow recovery of the rapid Na+ system. The phenomenon may be responsible for reported QRS prolongation and fatal ventricular arrhythmias encountered in patients receiving phenothiazines.


A bacterial cardiotoxin: thermostable direct hemolysin produced by Vibrio parahaemolyticus.


Selective effects of TTX and verapamil on the upstroke components of the action potential from the atrioventricular node.


Effect of low protein diet on cardiac function and ultrastructure of spontaneously hypertensive rats loaded with sodium chloride. The effects of low protein diet on cardiac metabolic and structural changes subsequent to an extremely high pressure load were investigated. Spontaneously hypertensive rats(SHR) were divided into four groups, and fed the following diets for four weeks: 1) Group A: regular diet (23% protein) and water, 2) Group B: regular diet and 1% saline, 3) Group C: low protein diet (10% protein) and water, and 4) Group D: low protein diet and 1% saline. Two weeks after the start of feeding, there was no significant difference in the left ventricular ultrastructures between the corresponding regular and low protein diet groups. Four weeks after, degenerative findings such as streaming of Z lines, dilatations of smooth endoplasmic reticulum, and disarrangements of myofilaments appeared Group D, while in Group B electron microscopic findings indicated hypertrophy. Incorporation of [14C] leucine into the myocardial myosin B in Group D was significantly low, with a resulting fall of the LVdP/dtmax per integrated isometric pressure (IIP) and a rise of the left ventricular end-diastolic pressure (LVEDP), as compared to that in Group C at four weeks after the start of feeding. These observations suggest that, in the heart with an extremely high pressure load, low protein diet hinders the development of hypertrophy to the load with resulting heart failure.


Increase in membrane conductance and positive inotropic action of hemolysin produced by Vibrio parahaemolyticus on rabbit myocardium. The effects of purified thermostable direct hemolysin from the cultured filtrate of Vibrio parahaemolyticus on both S-A node cells and atrial cells were studied electrophysiologically. Hemolysin caused the membrane conductance to increase, thereby causing membrane depolarization. The major ion responsible for this depolarization was Na+, but other ions, such as K+ and Ca2+, also participated. Positive inotropic action of hemolysin was observed, and the Na-Ca exchange mechanism was enhanced after hemolysin treatment.


Stopping of spontaneous beating of cultured mouse and rat myocardial cells by a toxin (thermostable direct hemolysin) from Vibrio parahaemolyticus. Low concentrations of thermostable direct hemolysin from Vibrio parahaemolyticus stopped the spontaneous beating of cultured mouse and rat myocardial cells. These low concentrations depolarized the maximal diastolic potential and inhibited the generation of action potential of cultured myocardial cells. The toxin lost its activity when preincubated with ganglioside, GT1 or GM1. GT1 was more effective than GM1. High concentrations of the toxin caused morphological damage of cultured mouse and rat myocardial cells, but did not stop the beating, or cause morphological damage of cultured chick myocardial cells.


Ischemic-like change produced in canine heart by atractyloside: a chemical model. Experiments were conducted in two canine models. Intracoronary infusion of atractyloside resulted in physiological, biochemical, and ultrastructural changes similar to those seen with ischemia. The fact that atractyloside, a specific inhibitor of adenine nucleotide translocase produces this change in vivo suggests that this inhibition in the key disturbance that initiates the metabolic derangements seen with ischemia. This chemical model seems to provide unique opportunities to study the possible factors that may reverse adenine nucleotide translocase inhibition and thus possibly prevent cell injury.


Factors accounting for growth and atrophy of the heart. Growth of the rat heart was induced by intrathoracic aortic banding, while atrophy followed hypophysectomy. During the early phase of hypertrophy, RNA concentration increased markedly. During the period of atrophy, RNA concentration was reduced. These changes in RNA concentration seemed to account for either the increased or the decreased rates of protein synthesis that was observed during hypertrophy or atrophy, respectively. Rates of protein degradation were unaffected during changes in heart size. These findings suggest that alterations in synthesis, processing, or degradation of RNA play a key role in modification of heart size.


Local pulsatile intramyocardial pressure (IMP) as a vector force: simultaneous measurements of IMP and tissue oxygen availability. A directional nonuniformity of the intramyocardial pressure (IMP) was observed in anesthetized open-chest dogs using a piezoelectric pressure sensor implanted in the left ventricular myocardium. The nonuniformity seemed to be attributable to the variation in myocardial fiber orientation. The IMP, which developed in a direction parallel to the base-apex line, was measured in the deep portion of the myocardium, simultaneously with local O2 availability. In some cases, IMP decreased quickly as soon as a decrement in O2 availability had proceeded only slightly as a result of ligation of the anterior descending artery (LAD). In other cases, the once decreased IMP recovered gradually even when O2 availability remained lowered by continuation of the ligation. These observations indicated that IMP had no direct dependency on O2 availability in the case of acute LAD.


Effects of anoxia and metabolic inhibitors on reaction of the fast sodium system. The effects of anoxia and metabolic inhibitors on inactivation and reactivation of the fast Na system of dog ventricular muscle fibers were studied by examining (dV/dt)max of the action potential upstroke. Anoxia for 60 min, dinitrophenol (DNP) (2.0 mM), and sodium azide (2.0 mM) depressed markedly steady-state (dV/dt)max at each membrane potential produced by changing [K]o. Anoxia tended to prolong recovery slightly, but DNP and azide changed little the time course of recovery of the (dV/dt)max, which was studied by applying double pulses to elicit action potential at different intervals. It is concluded that anoxia, DNP, and azide caused prominent depression of the maximum Na conductance but had less influence on its recovery kinetics.


Changes in intramyocardial oxygen tension and local electrograms produced by coronary occlusion.


Effects of L-lactate on glyceraldehyde-3-P dehydrogenase in heart muscle.


Species difference of mitochondrial respiration in experimentally induced heart failure. In guinea pigs with acute heart failure, a decrease in oxygen consumption (QO2) at state 3 and an increase in QO2 at state 4 were seen and the respiratory control index (RCI) was markedly lowered; ADP/O ratio was not affected. In rabbits with acute heart failure, both RCI and ADP/O ratio showed a statistically significant decrease. In dogs with acute and chronic heart failure, no impairment of respiratory function of mitochondria was seen. These conflicting results may be attributed to the difference in experimental conditions, namely, the species difference and the difference in methods to produce experimental heart failure. With special regard to species difference, it was confirmed that the RCI of myocardial mitochondria of healthy dogs was much lower than that of healthy guinea pigs. It is suggested that the difficulty of obtaining intact mitochondria may be responsible for the species difference in experimentally induced heart failure.


Transmural gradients in myocardial gas tensions in regionally ischemic canine left ventricle. Previous studies from this laboratory have demonstrated the usefulness of myocardial gas tensions as measured by mass spectrometry for the quantitative assessment of regional myocardial ischemia (Khuri et al., 1975a). Progressive increases in myocardial carbon dioxide tensions were noted when progressive reduction in coronary blood flow was created by means of a variable constrictor. The present study was designed to determine if changes in myocardial oxygen and carbon dioxide tension were greater in deep, compared to more superficial, myocardial layers. In eight anesthetized dogs, progressive reduction in circumflex coronary flow was associated with a progressive reduction in myocardial oxygen tension and a progressive increase in myocardial carbon dioxide tension and intramyocardial ST-segment voltage. Evidence of a transmural gradient in the severity of ischemia was present at all degrees of flow reduction. These results confirm the findings of previous metabolic studies, which demonstrated gradients in lactate and high-energy phosphates. Myocardial carbon dioxide tension, which can be monitored continuously by mass spectrometry, would appear to provide a useful means of quantitatively assessing changes in regional myocardial metabolism.


Hemodynamic and metabolic changes caused by regional ischemia in porcine heart. Myocardial ischemia was induced in the pig by reducing the left anterior descending coronary artery (LAD) flow to 26% of its control value. After one hour, the LAD was reperfused for 50 minutes. During ischemia and reperfusion, cardiac output correlated strongly with arterial levels of lactate and nucleosides (all r less than - 0.85). Myocardial inosine production seems to be a good marker of ischemia because it correlated very well with myocardial lactate production (r = 0.99).


Effects of acetate on myocardial contractility and epicardial ECG of isolated and perfused feline hearts. In isolated, perfused hearts, the effects of acetate on epicardial ECG and ventricular pressure were observed during various times of perfusion. Anions-Cl-and acetate-caused changes of ECG configuration, especially in the ST segments. These were different in Phase I (fast compartment) and Phase II (slow compartment).


Abnormal myocardial fluid retention as an early manifestation of ischemic injury.


Metabolic consequences of Sephadex-induced reduction of coronary flow in isolated rat heart. With a polysaccharide microsphere suspension (Sephadex), coronary flow of isolated perfused rat hearts was reduced by approximately 70%. During this Sephadex-induced ischemia, the energy charge and creatine phosphate content of the myocardial tissue dropped significantly, while total nucleoside and lactate release from the heart increased. During hypoxia (30% O2), changes in high-energy phosphate content and lactate and nucleoside release were similar to the changes induced by ischemia. During hypoxia, coronary flow rate was increased by 47%. Thus, Sephadex-induced reduction of coronary flow could be a useful model in studies of metabolic change during ischemia.