Ultrastructural study of leukemic cell phagocytosis using the myeloperoxidase reaction. The phagocytosis (in the absence of serum factors) of zymosan particles by peripheral leukocytes isolated from ten patients with acute leukemia (AMbL, AMoL, AMML, AUL, ALL and CML-BC) was studied at the electron microscope. An evident phagocytic activity was observed only in the cells in which cytochemical and ultrastructural features suggested that the blast elements belonged to the monocytic series. However, no phagocytosis by unclassifiable leukemic blasts was observed, even though they had some submicroscopic characteristics of the monocytic series. These findings suggest that phagocytic capacity develops during the course of cell differentiation, becoming striking only when the blast cell acquires the ultrastructural features of the pro-monocytic stage. Using the myeloperoxidase reaction, this study also demonstrates a morphological alteration in the degranulation process after the ingestion of zymosan particles in both the blasts and the mature PMN cells of leukemic patients. This defect could be related to the susceptibility to severe infections usually found in subjects with hematological malignancies.


Early detection of ductal breast cancer: the diagnostic procedure for grouped microcalcifications. Mammography and xeroradiography for grouped microcalcifications are considered the most effective diagnostic methods to detect occult breast carcinoma. Radiography must direct the surgeon to excise the nonpalpable area. The removal of the tissue with grouped microcalcifications must be confirmed by intraoperative radiological control. The histologic preparation must be guided by radiographic controls. Tissue with calcific deposits is examined by step sections. The diagnostic success depends upon the cooperation between the radiologist, the surgeon, and the pathologist. Our results from 1964 to 1977 have shown a frequency of 14.4% of occult carcinoma. Ductal or lobular carcinomata in situ have been diagnosed in 8.9%. In 9.9% of the patients, cystic disease with severe and atypical proliferations has been encountered.


Early detection of ductal breast cancer: the diagnostic procedure for pathological discharge from the nipple. Pathologic discharge from the nipple may be the only symptom of an early stage of carcinoma. Galactography is then the diagnostic method of choice to locate intraductal, nonpalpable lesions. The technique of galactography, the adequate surgical approach of pathologic galactographs (milk-duct segment resection), and the appropriate histological work-up of the surgical specimen are demonstrated. We report on 1918 galactographies in 1363 women with pathological discharge. In only 427 cases was a milk duct segment resection necessary (31.4%). In 8.5%, we found invasive intraductal cancer and in 2.9% ductal carcinomata in situ. Only 1 patient with breast cancer had axillary metastases. Extensive intraductal solid, papillary or adenomatous proliferations were found in 11.9% of the patients with excision. In 46.7% of the patients, papillomas were excised, a definitive treatment for this process. The supposition for success in the early diagnosis of cancer is close teamwork among the radiology, surgery and pathology services: the diagnostic result depends upon this. We attribute our yield of exact diagnosis to a very sophisticated histological work-up. We believe that this is necessary to avoid diagnostic failures.


Lobectomy with bronchoplastic procedures for lung cancer. From April 1970 to October 1977, 19 patients with lung cancer of the upper lobar bronchus orifice underwent radical lobectomy with major bronchus resection. Sleeve lobectomy was accomplished in 11 cases and wedge lobectomy in the remaining 8. The length of the free bronchial margin in the surgical specimen was less than 1 cm in 3 cases, but limited pulmonary reserve did not allow pneumonectomy. Squamous carcinoma was diagnosed in 14 patients, adenocarcinoma in 2, oat-cell carcinoma in 2, and large cell carcinoma in one. Most cases (70%) were pathological stage I. There was one operative death due to anastomotic leakage (5%), and another patient required pneumonectomy completion. Of 13 patients with non oat-cell carcinoma and adequate bronchial resection, none had local recurrence: 3 patients developed distant metastases, and 10 are alive and disease-free after a follow-up period ranging from 16 to 104 months. The authors conclude that in selected lung cancer patients lobectomy with bronchoplastic procedures is superior to pneumonectomy for tissue sparing advantages.


Unusual morphologic features of a primary liver neoplasm. Light and electron microscopy study of an unusual primary liver neoplasm observed in a 36-year-old woman. By light microscopy the tumor, initially detected by a needle biopsy and later surgically removed, was mainly composed of spindle-shaped and multinucleated cells which were similar to rhabdomyoblasts. These cells, however, showed ultrastructural features of hepatocytes.


[Physicochemical and kinetic characteristics of myeloperoxidase of cow neutrophilic leucocytes]. Myeloperoxidase (MPO) isolated from granular fraction of cow peritoneal neutrophils had the RZ value equalled to 0.79. The absorption spectrum of MPO possesses characteristic bands at 430, 500, 570, 625, 690 nm. The molecular weight and sedimentation constant of the enzyme are 135.000-140.000 Daltons and 7.9S, respectively. The data obtained demonstrate the high thermostable property of MPO. Investigations of the pH optimum and effects of varying H2O2, o-dianizidine and MPO concentrations are described.


[Activity of certain redox enzymes of rat liver mitochondria at different levels of dietary vitamin E]. Taking into account the found earlier relation of vitamin E to the ubiquinone functioning and metabolism, the authors studied the enzymic activity of succinate dehydrogenase, NADH-dehydrogenase and cytochrome-c-oxidase--coenzyme Q binding sites of the respiratory chain of the rat liver mitochondria. The experiments were carried out with female rats who received a vitamin-E-deficient diet for 6 months. The enzymic activities and the ubiquinone content in the liver mitochondria of these animals are shown to be considerably lower as compared to the animals received a vitamin E diet; alpha-ocopherol, alpha-tocopheronolactone and ubiquinone 3h after administration manifest a clearly pronounced normalizing effect relative to both the enzymic activity and the ubiquinone content. An assumption is advanced that the effect of alpha-tocopherol and its metabolite is associated with controlling the level of functionally active ubiquinone in the mitochondria. Other mechanisms of the membrane-bound enzymes control by the compounds under study are also discussed in connection with the alpha-tocopherol effect on the mitochondrial membranes.


[Use of a selective sulfite-reducing medium for the isolation of Clostridium perfringens]. The effect of sulphite-reduction in 33 sample bacterial strains  was tested. With regard to the capacity of reducing sulphite in modified sulphite-reduction media in a wide scale of bacterial strains the possibility of an application of selective media with an addition of various concentrations of antibiotic solutions was checked. A concentration of 750 microgram of D-cycloserine per 1 ml of the sulphite-reduction medium appeared to be the most advantageous for the isolation and detection of sulphite-reductive clostridia, above all of Clostridium perfringens. This concentration ensured also a sufficient inhibition of undesirable bacteria without any affection of the growth and capacity of Clostridium perfringens to reduce sulphite in the applied medium.


[The effect of halothane anesthesia on the function of the adrenal cortex and some metabolites in the blood plasma of pigs not susceptible to malignant hyperthermia]. The effects were investigated of a 25-minute inhalation of halothane with oxygen on three to four months old pigs of the Large White breed. Symptoms of malignant hyperthermia did not occur. The actual total anesthesia, which causes slight hypoproteinemia, hypoglycemia and hypocholesterolemia without significant changes in the content of non-esterified fatty acids (NEFA) and urea, induced only a  slight increase of circulating 11-hydroxycorticosteroids (11-OHCS). The combination of anesthesia with castration of gilts or barrows significantly increased the concentration of 11-OHCS but did not reach the level recorded after the application of ACTH. The higher levels of 11-OHCS were accompanied by higher concentrations of NEFA and glucose. The treatment of the animals lasting half an hour prior to inhalation of halothane at maximum doses or one hour in the control unanesthetized pigs produced an effect, mainly on the 11-OHCS concentration and on the activity of creatine kinase in the plasma. The results indicate that the adrenocortical response to the effect of halothane is not stronger than the response to simple handling connected with excitement and muscular activity of the animals. Therefore there is no reason of considering halothane anesthesia as a factor causing great stress and pigs which in its course do not respond with malignant hyperthermia as animals insensitive to stress. The aptness of denotation of clinical manifestations of genetically defective muscles in pigs is discussed.


[Modern concepts of the metabolism and genetic polymorphism of collagen (a review)]. The data on structure, biosynthesis, post-synthetic modifications, extracellular development and degradation of collagen are briefly reviewed. Role of main enzymes, participating in collagen turnover, is discussed. Isoforms of the protein are considered in details. The importance of studies on collagen proteins is noted in connection with investigations on morphogenesis, cell differentiation, regeneration.


[The protein-synthesizing apparatus in chemically- and virus-induced tumors]. The description is given of the distribution of poly(A)-containing mRNA in different subcellular fractions (nuclei, mitochondria, free- and membrane-bound polyribosomes) from the normal tissues hepatoma MD and chick Rous. The amount of poly(A)-RNA is found to increase in all tumor cell fractions, but free polysomes.


[Regulation of Na, K-ATPase activity in the rat brain by noradrenaline]. Effect of noradrenaline on activity of Na, K-ATPase from heavy microsomes of rat brain was studied in vitro; the effect of noradrenaline was shown to be dose dependent. The maximal activation of the enzyme was observed at 5.10(-5) M concentration of the catecholamine; an increase of its concentration above 5.10(-4) M inhibited Na, K-ATPase. beta-Adrenolytic drug propranolol /1.10(-5) M/ did not affect the noradrenaline action. At the same time, an alpha-adrenolytic drug phentholamine /1.10(-5) M/ decreased down to zero the activating effect of the catecholamine, but did not affect the inhibitory effect of noradrenaline at high concentrations /1.10(-3) M/. Effects of selective alpha-adrenomimetic compound phenylephrine were also studied. The data obtained suggest that the activating effect of noradrenaline on Na, K-ATPase from rat brain is mediated through alpha-adrenoreceptors.


[Characteristics of thiamine diphosphate metabolism in the liver in acute hypobaric hypoxia]. Rates of thiamin diphosphate accumulation in rat liver tissue and its subsequent depletion were decreased within the first hour after vitamin B1 administration into the animals with oxygen insufficiency. Hypoxia apparently inhibited simultaneously the processes of thiamin diphosphate biosynthesis and degradation, as a result of which content of the coenzyme was unaltered in tissues under conditions of usual intake of thiamin. The data obtained suggest that the activity of enzymes, participating in thiamin diphosphate turnover and estimated in vitro do not reflect their real functioning in the whole organism. Importance of ATP, ADP and pyridine nucleotides is considered as regulating agents of enzymes participating in thiamin diphosphate turnover under hypoxia.


[Structural characteristics of plasma lipoproteins studied with spin-labelled fatty acids]. As shown by means of electrone paramagnetic resonance of spin probes, a magnitude of ordering of lipoprotein molecular arrangement both internal and external parts of molecule was decreased in a following sequence: high density lipoproteins greater than low density greater than very low density lipoproteins. Permeability towards a reducing agent - ascorbic acid - was altered similarly in the lipoprotein surface monolayer.


[Breast angiography in cancer].

[Breast angiography in cancer]. The roentgenological features of the blood supply of mammary glands removed for cancer are analysed. It was found that the tumor angioarchitectonics is dependent on the kind of growth and histological structure. Some angiographic changes were noted in the vascular network of the mammary gland and tumor, arising under the effect of the radical preoperative telegamma-therapy.


[Changes in the indices of fat metabolism in obesity during physical activity, on a reducing diet and during meterological influences]. Clinical experimental investigations were carried out of 96 patients with light, nutritional-metabolism forms of obesity under the conditions of a moderately high mountain (1870 m above the sea level). After a treatment course of 20--25 days including: reducing diet, physical loading and cool sun-air exposure, a significant decrease of a wide spectrum of lipid indices was found, namely: total lipids, triglycerides, total cholesterol, phospholipids and beta-lipoproteins. A significant correlation was established between cholesterol decrease and beta-lipoproteins with body weight reduction over 3 kg and the high initial values of the indices. With a single physical loading in the patients with obesity -- the elevation of NEFA does not considerably differ from that of the control group with neurasthenia. The glucose loading is admitted not to indice manifested lipogenesis with the physical loading and cool sun-air exposure.


[Effect of small amounts of methylmercury on embryogenesis and certain biochemical indices]. It was established in experiments on Wistar rats that oral methyl mercuric iodide at doses of 0.85, 0.64, 0.42 and 0.21 mg/kg daily exerts no teratogenic or embryotoxic action on the progeny. However, administration of methyl mercuric iodide at doses of 0.85 and 0.64 mg/kg produced in females a significant decrease in the SH-group content in renal and cerebral tissues, in the activity of acetylcholine esterase in the cerebellum (by 18-48%) and a rise in the activity of glucose-6-phosphatase in the kidneys (by 48-70%). The doses of 0.42 and 0.21 mg/kg administered at various periods of pregnancy, did not exert any effect on the body of females under experimental conditions. These findings are supported to a certain measure by the results of examination of a 2-month progeny of the animals given 0.21 mg/kg methyl mercuric iodide daily. The biochemical parameters of these animals did not differ from those of the control ones.


Clinical and virologic course of herpes simplex genitalis. The clinical and virologic course of herpes simplex genitalis in women and men was examined in order to identify measurements useful in antiviral trials. Factors influencing the clinical course included initial disease versus recurrent disease, wet-skin versus dry-skin lesions, female versus male sex. Women with initial genital herpes had higher mean peak lesion virus titers than those with recurrent disease (10(4.5) pfu compared with 10(2.5) pfu) and excreted virus longer (13 to 15 days compared with 6 to 8 days). Men with recurrent lesions had higher mean peak virus titers than women (10(4.0) pfu compared with 10(2.5) pfu), but the duration of virus excretion was shorter (three to four days compared with six to eight days). There was pronounced variation in the clinical and virologic course of recurrent lesions among different patients and even within the same patient. These observations indicate several difficulties that must be considered in conducting careful antiviral trials in patients with herpes simplex genitalis.


Severe neutropenia in infectious mononucleosis. Mild neutropenia is a well-known concomitant of infectious mononucleosis caused by the Epstein-Barr virus (EBV) occurring in the first weeks of illness. However, severe neutropenia (less than 200 polymorphonuclear leukocytes per mul) is not generally regarded as a complication of infectious mononucleosis. Three patients were seen with severe neutropenia and EBV infection, and an additional eight cases were found in the literature. In two of the latter cases the neutropenia was fatal. In the 11 cases the severe neutropenia began 14 to 40 days after illness and usually lasted for three to seven days. At the time of severe neutropenia, studies of marrow specimens showed increased proportions of promyelocytes and myelocytes. Our data suggest that EBV infection is the proximate cause of the severe neutropenia in some patients with infectious mononucleosis and that in such cases close observation and early treatment of suspected superinfections is necessary.


[Psychosomal dwarfism with reversible growth hormone deficiency (author's transl)]. The diagnosis of psychosocial dwarfism in a 9 year-old boy with severe growth retardation (-6 1/2 standard deviations) was deduced from the typical history. The bone age was severely retarded and in the first days after admission a deficiency of growth hormone and other pituitary hormones was established. The change in environment per se led to a spontaneous reversal of the growth hormone deficiency within a short time. A rapid catch up growth was observed over the subsequent 2 1/2 years, as well as a normalisation of the psychological retardation.


[Diagnostic and prognostic value of scintigraphic methods in cardiology (static methods)]. The diagnostic possibilities of myocardial scintigraphy in cardiology are discussed. As an invasive technique the perfusion imaging is described. The valuation of the doubleisotope-scintigraphy for better diagnostic examination during coronary arteriography is accentuated. The 201-Thallium-scintigraphy as a non invasive method is debated. The diagnostic value of the 201-Thallium stress scintigraphy is shown. The similarity of ergometric and pharmacologic provocation with Dipyridamol is represented. In the following the reliability and the specificity of the infarct scintigraphy with 99m-Technetium-pyrophosphate as "hot-spot-technique" is discussed. The follow-up scintigraphy allows to draw prognostic inferences about the further course of a myocardial infarction. The scintigraphic findings were correlated to haemodynamic measurements. Scintigraphic examinations in patients with stable angina pectoris without infarction revealed correlations between the uptake of the tracer in the myocardium and a disturbed left ventricular function. A chronic deficiency in the blood supply of the myocardium is discussed as the cause of the tracer uptake.


[Sleep and wake-cycle as circadian rhythm (author's transl)]. Sleep-wake cycles are closely tied with the circadian period of other bodily functions. Optimal synchronization is essential for troublefree alterations of sleep and wake periods. The neurophysiologic control areas are in various of the brain are far apart from each other for which reason a great variety of possible causes for sleep difficulties do exist. It has been shown recently that spezific mono-aminergic and cholinergic systems in the brain stem do control sleep. A case study will show that it is possible to improve sleeping difficulties with the serotonin precursor L-Troyptophan.


Vasoactive intestinal polypeptide (VIP) as a mediator of the watery diarrhea syndrome. Evidence that VIP is the principal humoral mediator of the watery diarrhea syndrome includes: (a) actions of VIP in experimental anaimals parallel the clinical manifestations of the syndrome; (b) infusions of VIP induce watery diarrhea in intestinal loops of dogs and a picture resembling the clinical syndrome in pigs, at circulating levels of the peptide similar to those observed in human disease; (c) most patients with the watery diarrhea syndrome and underlying tumors have elevated plasma levels of VIP; (d) in those patients in whom pre- and postoperative measurements were made, plasma VIP levels fell to the normal range with removal of the tumor and relief of the diarrhea; and (e) extracts of such tumors are rich in VIP-immunoreactivity and VIP-like biologic activity. A few patients with the syndrome have been reported to have normal plasma VIP levels, and it is possible that other humoral agents (such as pancreatic polypeptide, prostaglandins) may contribute to the production of the diarrhea.


Heterotransplantation of human gastric carcinomas into nude mice. A total of 33 specimens of human gastric carcinoma were used for transplantation into nude mice. Initital tumor "take" was accomplished in 15 of the 33 tumors, and the transplantability rate was 45.5%. Transplantability correlated with histologic type, but not with clinical stage or Borrmann's classification. The transplantability rate of differentiated carcinomas, such as well-differentiated tubular adenocarcinoma, moderately differentiated tubular adenocarcinoma, and papillary adenocarcinoma was greater than that of poorly differentiated tumors, such as poorly differentiated adenocarcinoma and mucinous adenocarcinoma. The growth patterns of transplanted tumors were divided into 3 types: rapid, slow, and persistent. There were no specific relationships between growth pattern and histologic type. All histologic types, except signet ring cell carcinoma, could be transplanted serially. Tumor growth became rapid after serial transfer. However, the original histology of these tumors was unchanged. No invasion or metastases were encountered. Intraperitoneal injection of a tumor cell suspension, prepared from subcutaneous transplants of a poorly differentiated adenocarcinoma of Borrmann type III, grew in an ascites form, with invasion and metastasis. Ascitic fluid accumulated within 3--6 weeks after injection. Subsequently, intravenous injection of ascites fluid produced metastases in nude mice. The histology of the subcutaneous tumor was similar to that of the original tumor from the patient.


Effects of dibutyryl cyclic AMP and cytochalasin B on cultured human glioma cells. Cultured human glioma cells (138 MG) exposed to dibutyryl cyclic AMP (dbc-AMP; 0.1--5 mM) attained an arborized shape with thin processes extending from a rounded cell body. Cytochalasin B (CB; 1--1 muM) induced similar morphological changes. The processes in both dbc-AMP and CB treated cells were formed by retraction of the cell margin. Colchicine (1muM) completely and liver treated phalloidin (0.1 mg/ml) partially inhibited the morphological alterations induced by dbc-AMP and CB. Dbc-AMP was found to arrest cell movement, cell division and uptake of 2-deoxy-D-glucose. CB has the same effects but was more potent. The effects of dbc-AMP and CB could be due to interference with a common cellular structure, e.g. microfilaments.


[Determination of total mercury in fish by means of a digestion method with HNO3/HClO3/HClO4 which does not lead to low results (author's transl)]. A digestion method for determination of total mercury in fish is described, which is economical, simple and reliable. Digestion is carried out by wet ashing with HNO3/HClO3/HClO4 in the presence of chloric acid. The procedure can be carried out at normal pressure in test tubes without suffering losses of Hg. The determination itself is carried out by the well known cold vapor atomic absorption method. This method was worked out specially for the regulatory control of fish, but can probably be used for related problems.


[Rotavirus diarrhea in young infants].

[Rotavirus diarrhea in young infants]. Clinical and serological studies were made in 62 children with various purulent infections, who were placed in the same hospital unit. The precipitation test with the antigen of monkey virus SA-11 showed that diarrhea in 32 of these children was etiologically linked with rotaviruses. The diagnostic titers of virus antibodies were detected in children having diarrhea 5.5 times as frequently as in the control group. The characteristic feature in the clinical picture of this diarrhea was the absence of the severe form.


[Nervous system pathology in uterine myomas]. Clinico-physiological examinations of 218 patients with uterine myoma were carried out using the methods of electromyography and rheopgraphy. The following syndromes were revealed: a myopathic and a radiculoalgic ones; the syndrome of the external dermal femoral and the femorogenital nerves. The pathogenesis of these syndromes consists in compression of the lumbar arteries and the minor pelvis ganglia by the enlarged uterus. On condition of timely treatment including surgical ablation of the myoma the prognosis of the neurological disturbances is, as a rule, favourable.


[Clinical and electromyographic characteristics of syndromes of pathologic muscular fatigue of the myasthenic type]. On the basis of the follow-up study of more than 2000 patients with different forms of pathological muscular fatiguability of the myasthenic type, the authors emphasize the heterogeneity of this group of patients. The detailed study of clinical and electromyographic picture permitted them to distinguish the following clinical forms; myasthenia, myasthenic syndrome in terminal polyneuropathy, the myastheniapolymyositis complex, myasthenic syndrome of Lambert--Etone type.


Na,K-ATPase in excitation-contraction coupling of vascular smooth muscle from cattle. Lowering the extracellular K+ content from 6 to 0.6 mM causes a rise, and elevation from 6 to 8.5 mM a fall of 45Ca++ efflux from the vascular smooth muscle cells of the arteria carotis communis of cattle. In contrast, a level of 17 mM K+ has no influence. Removal of extracellular calcium does not block these effects. 10(-4) M ouabain also induces a rise in Ca++ efflux, additional potassium reduction then being without effect; 10(-9) M ouabain is of no influence. The 45Ca++ efflux kinetics correlates with the activity of the isolated Na,K-ATPase. Tonus increases of the vascular strips by 10(-4) M ouabain and potassium deficiency cannot be blocked by 4 mM lanthanum or removal of extracellular calcium. Unlike sodium, potassium stimulates the active Ca++ binding and the activity of the Ca-ATPase of the microsomal fraction. The ative Ca++ binding of the mitochondria is stimulated by both ions. It is postulated that the activity of the plasma membrane Na,K-pump is able to regulate the tonus of big arteries through alteration of Ca++ storage processes.


A thermodynamic-kinetic analysis of the cytochrome P-450 heme pocket. Spin state changes in the iron center of cytochrome P-450 during the catalytic cycle suggest alterations in the heme environment that insure proper substrate binding, an increase in redox potential, the formation of an active Fe-O complex, and the attack on the substrate. We used the spin state changes of the iron following physico-chemical perturbations, as an intrinsic probe of discrete changes around the heme, or of larger ones in the protein conformation. These environmental perturbations included temperature, solvent, substrate, and ionic environment. Aqueous and hydro-organic buffers provide complementary data and interpretations; the mixed solvent accommodates temperatures suitable for direct reaction rate measurements and amplified low to high spin transition. The results suggest that the group determining the heme spin state is influenced by the electrostatic potential created by several negative charges near the heme; the modulation of the spin state by various factors reflects the modulation of the electrostatic potential and of the internal paH value. Conformational changes of the whole protein are also indicated by the large entropy terms and their variation with experimental conditions.


[Pyramidal approaches to surgical treatment of pathologic formations of the cerebellopontine angle and pyramids of the temporal bone]. Pyramidal approaches are sufficiently effective, cause little traumas, and are easily tolerated by the patients. They are indicated in surgical management of diseases of the cerebellopontine angle such as tumors (neurinoma of the auditory nerve, cholesteatoma), cystic arachnoiditis, tumors of the temporal bone pyramid (glomus tumors, cysts), compression of the facial nerve in mechanical trauma of the pyramid, etc. The choice of the variant of the pyramidal approach to the pathological focus should be guided by the following main criteria: localization of the process and the extent of its spreading; condition of the facial nerve; condition of the auditory function.


Cytochrome P-450cam and putidaredoxin interaction during electron transfer. Cytochrome P-450cam, the bacterial hemeprotein which catalyzes the 5-exo-hydroxylation of d-camphor, requires two electrons to activate molecular oxygen for this monooxygenase reaction. These two electrons are transferred to cytochrome P-450cam in two one-electron steps by the physiological reductant, putidaredoxin. The present study of the kinetics of reduction of cytochrome P-450cam by reduced putidaredoxin has shown that the reaction obeys first order kinetics with a rate constant of 33 s-1 at 25 degrees C with respect to: 1) the appearance of the carbon monoxide complex of Fe(II) cytochrome P-450cam; 2) the disappearance of the 645 nm absorbance band of high-spin Fe(III) cytochrome P-450cam; and 3) the disappearance of the g = 1.94 EPR signal of reduced putidaredoxin. This data was interpreted as indicative of the rapid formation of a bimolecular complex between reduced putidaredoxin Fe(III) cytochrome P-450cam. The existence of the complex was first shown indirectly by kinetic analysis and secondly directly by electron paramagnetic resonance spectroscopic analysis of samples which were freeze-quenched approximately 16 ms after mixing. The direct evidence for complex formation was the loss of the EPR signal of Fe(III) cytochrome P-450cam upon formation of the complex while the EPR signal of reduced putidaredoxin decays with the same kinetics as the appearance of Fe(II) cytochrome P-450. The mechanism of the loss of the EPR signal of cytochrome P-450 upon formation of the complex is not apparent at this time but may involve a conformational change of cytochrome P-450cam following complex formation.


Spin state transitions of liver microsomal cytochrome P-450. Spin state transitions of membrane-bound cytochrome P-450 were investigated by difference spectrophotometry using the 'D'-charge transfer absorbance band at 645 nm as a measure of the amount of hemin iron present in the 5-coordinated state. The magnitude of the 'D'-absorbance band in the absence of exogenous substrates, e.g., the concentration of native high spin cytochrome P-450, was evaluated from the difference in absorbance at 645 nm between ferric cytochrome P-450 and the carbon monoxide derivative of the pigment in its ferrous state. The contribution of the native high spin species to the total cytochrome P-450 content of microsomes was calculated to be between 40% and 65% after induction with phenobarbital and polycyclic hydrocarbons, respectively. Up to 80% of the cytochrome P-450 was found to be present in the high spin state after the addition of exogenous substrates. Further, the steady state concentrations of high spin cytochrome P-450, observed in the presence of reduced pyridine nucleotides, suggest that the rate limiting step for microsomal mixed function oxidation reactions is variable and dependent on the substrate under investigation.


Quantitative analysis of the spin equilibrium of cytochrome P-450 LM2 fraction from rabbit liver microsomes. The LM2 fraction of cytochrome P-450 from phenobarbital induced rabbit liver microsomes in the presence and in the absence of substrate (benzphetamine) is shown to be a thermal equilibrium of a high spin (S = 5/2) and a low spin (S = 1/2) state each of which exhibiting its individual optical basic spectrum with the Soret maxima at 387 nm and 417 nm for the high spin form and the low spin form, respectively. The equilibrium constants and thermodynamic parameters describing the spin transition and the substrate binding have been evaluated from the temperature and substrate difference spectra. These two interacting equilibria are presented in terms of a thermodynamic model, which provides a quantitative description of the relationship between the substrate binding and the spin equilibrium. From kinetic experiments it is concluded that the spin equilibrium is an electronic one and is not caused by iron ligand dissociation.


The importance of the spin equilibrium in cytochrome P-450 for the reduction rate of the heme iron. Based on the existence of a spin equilibrium of P-450 LM a thermodynamic model including the first two elementary steps has been developed. This model has been compared with a modified model of P-450 CAM first presented by SLIGAR [1]. For the quantitative description of the model of P-450 LM seven necessary constants have been calculated. In the model a 5-coordinated ferric heme complex as intermediate and precursor of the reduced state is included. The model is based on experimental data of the spin equilibrium and the binding of benzphetamine to P-450 LM2, on the redox potential of P-450 LM and further on the assumption of a structure of the precursor well adapted to reduction. The dissociation constant (KD) of the 6th heme iron ligand is calculated to be 0.03 meaning that only 3% of the high spin ferric state are 5-coordinated. From the thermodynamic parameters of the model the sequential order of the reaction process can be derived and conclusions can be drawn on possibly reaction controlling steps. Experimentally it is shown that the substrate induced spin shift is correlated with the reduction rate. This has been shown to be valid in dependence on the degree of saturation with only one substrate as well as with different substrates indicating that independent of the individual substrate only the shift of the spin state controls the reduction. From the experiments as well as from the model it can be concluded that P-450 works in the first reaction step in two states (different in structural and functional properties) being in equilibrium with each other. Different affinities of these two states towards the substrate shift the equilibrium. This shift increases the concentration of the reducible intermediate and by this the reduction as second elementary step is regulated.


Activated forms of oxygen in the metabolism of xenobiotics catalyzed by cytochrome P-450. The Cu(Lys)2 complex is shown to be an effective inhibitor of the oxidation of 3,4-benzpyrene and aniline in microsomes. Apparently, this is due to the dismutation of superoxide radicals involved in the metabolism of these compounds. Possible interactions between Cu(Lys)2 and model membranes have been studied using spin probes. A model has been suggested according to which Cu(Lys)2 "builds" in the membrane hydrocarbon regions.


Comparison of spectral properties of 3-MC induced cytochrome P-448 from rabbits and rats. EPR and optical spectral properties of cytochrome P-488 from 3-methylcholanthrene-induced rabbits are compared with those of rats. In the EPR spectra at 77K and in the optical absorption spectra at room temperature a considerable temperature independent high spin content of the rabbit cytochrome is observed which has been estimated to about 55% by titration with n-octylamine. On the other hand the high spin content of the rat cytochrome depends strongly on temperature and amounts to about 6% at 5 degrees C and to about 35% at 34 degrees C. The binding of substrates and ligands to the rabbit cytochrome as well as its reduction by sodium dithionite are slower as compared with the rat cytochrome but also with phenobarbital-induced cytochrome P-450 from rats and rabbits. Contrary to the 3-methylcholanthrene induced cytochrome P-448 from rats, that from rabbits does not bind 3-methylcholanthrene. A particular protein structure establishing the high spin state and an absent binding site for type I substrates is assumed to be responsible for these and other peculiarities of cytochrome P-448 from 3-methylcholanthrene-induced rabbits.


Comparison of microsomal and solubilized monooxygenases from rat and rabbit by proton magnetic relaxation. The paper presents results of a comparative study of the haem environment, by proton magnetic relaxation, in P-450 and P-448 monooxygenases from rat and rabbit, induced by phenobarbital and 3-methylcholanthrene, in both species. It was established that the method yields information on the accessibility of the haem iron for solvent molecules (protons), both in microsomes and in solubilized samples of various degrees of purification, i.e. association. The state of micelles in the solutions does not alter the haem iron accessibility. A slight difference was found for the microsomes suspended in a phosphate vs. pyrophosphate buffer, but this is without any consequence with regard to the species and form differences. The correlation time for the highly purified LM2 fraction of rabbit P-450 could not be determined more precisely than before for a sample of lower purity, because the relaxation rates are frequency independent. The correlation time for the rat P-448 monooxygenase was determined by dispersion measurements to be (4.1 +/- 0.4) x 10(-11) s. It was found that the PMRx behaviours of rabbit and rat monooxygenases are more alike in microsomes than in the partially purified solubilized form. The solubilization produces a pronounced alteration of the PMRx temperature dependence only for the rat 3-MC induced monooxygenase P-448. For the P-450 form the haem iron becomes less accessible on solubilization, both for the rabbit and the rat liver monooxygenases, whereas in case of rat liver P-448 the accessibility is considerably enhanced on solubilization. There is a substantial structural specificity of the haem environments from the two animal species, the one from rat being tighter. The reduced, NO-bound rabbit liver monooxygenase was studied also, but the results are not yet conclusive, except the fact that the unpaired spin from NO is thoroughly shielded from the solvent compared with the haem iron from the original sample. The following series of increased haem-iron accessibility emerges from the PMRx studies known so far: rat (P-448) less than rabbit (P-448) less than rat (P-450) less than rabbit (P-450) in microsomes, and rabbit (P-448, with 3-MC bound?) less than Pseudomonas putida (P-450) rat less than (P-448), less than rat (P-450) less than rabbit (P-450) from solubilized samples. For the latter, it appears that increased enzymic specificity goes along with a closing of the haem cleft.


Stereochemical properties of the binding site of liver microsomal cytochrome P-450 as studied by substrate analogous spin labels. For the characterization of the substrate binding site optical and EPR measurements with spin labelled substrates on solubilized and pure cytochrome P-450 were performed. Analogously to the unlabelled derivatives spin labelled n-alkylamines and isocyanides with different chain lengths are type II substrates. The Ks-values evaluated from optical (P-450 = 1.98 . 10(-6) M) and ESR (P-450 = 1.98 . 10(-4) M) measurements are very similar indicating no concentration dependences. Contrary to the unlabelled n-alkylamines the spin labelled compounds show an affinity almost independent of the chain lengths. The SL-substrates with a short distance between the functional group and the NO-group bound to P-450 induce pronounced changes of the ligand field of the heme iron and a large broadening of the signal of the immobilized nitroxide indicating intensive interactions between the unpaired electron of the nitroxide group and the paramagnetic heme iron. Elongation of the alkyl chains results in spectra of the Fe3+ complexes with only slight modification and a remained unbroadened signal of the immobilized nitroxide. The binding of the substrate through their functional groups together with a 1:1 stoichiometry of the P-450 SL-IC-complex give evidence for the same binding site in the near vicinity of the heme iron.


Model systems for the coordination chemistry of cytochrome P-450. Studies on model complexes have supported the presence of a mercaptide as the fifth ligand of cytochrome P-450 monooxygenases. When alcohol or thiol ligands are added to the sixth coordination position of a five-coordinated 4-nitrobenzene thiolate complex of FeIII protoporphyrin IX dimethyl ester chloride low spin complexes with optical and EPR-spectra very similar to cytochrome P-450 are obtained. From a comparison with all ligands of cytochrome P-450 and the model complexes it is concluded that a hard ligands must occupy the sixth coordination position of cytochrome P-450. An imidazole group is less likely, also in view of the ligand field parameters. The significance of the fifth and sixth ligand of cytochrome P-450 is discussed with respect to the monooxygenase mechanism.


Isolation and characterization of cytochrome P-450meg. The cytochrome P-450-dependent steroid 15 beta-hydroxylase system from Bacillus megaterium has been resolved into three components, 1) a NADPH-specific, FMN-containing flavoprotein reductase, molecular weight 55-60 000; 2) an iron-sulfur protein, molecular weight 13,000 and 3) cytochrome P-450meg, molecular weight 52,000. The cytochrome component has been purified to homogeneity, as judged by SDS-polyacrylamide gel electrophoresis and isoelectric focusing in polyacrylamide gel, and its amino acid composition has been determined. Cytochrome P-450meg has a pI of 4.9, a Stokes radius of 27 A and a sedimentation constant of 3.3 S. Electron paramagnetic resonance and optical spectra are typical of a low-spin cytochrome P-450. The fluorescence spectrum is indicative of a tryptophane residue in a relatively non-polar environment. In recombination experiments, the electron flow was shown to proceed from the reductase via the iron-sulfur protein to the cytochrome. It is also possible to exchange the different components of the mitochondrial 11 beta-hydroxylase system from bovine adrenals for corresponding components in B. megaterium. Substrate specificity studies indicate that only steroids with a 3-oxo-delta 4-configuration are hydroxylated by the B. megaterium hydroxylase system. When oxidizing agents were used, hydroxylation occurred both in positions 15 alpha and 15 beta. Further substrate specificity studies have shown that aniline and imipramine can function as substrates for the bacterial system.


Oxycytochrome P-450: its breakdown to superoxide for the formation of hydrogen peroxide. Four different experimental studies are described which were designed to evaluate the role of oxycytochrome P-450 in the formation of superoxide anions and hydrogen peroxide. The use of lipophilic copper chelates with superoxide dismutase like activity revealed that the primary site of interaction of these agents is related to the inhibition of the flavoprotein. NADPH-cytochrome P-450 reductase. Measurements of the proton assisted nucleophilic displacement of superoxide from oxycytochrome P-450 by high concentrations of sodium azide indicated an increase in the rate of hydrogen peroxide formation concomitant with the inhibition of the N-demethylation of ethylmorphine. Studies on the effect of NADH on the rate of hydrogen peroxide formation during NADPH oxidation by liver microsomes failed to reveal a stimulatory or synergistic effect in a manner analogous to results obtained during the cytochrome P-450 dependent oxidation of substrates such as ethylmorphine. These results suggest that hydrogen peroxide formation may not require the reduction of oxycytochrome P-450 to peroxycytochrome P-450. Measurements of the reduction of succinylated cytochrome c using purified cytochrome P-450 and the flavoprotein, NADPH-cytochrome P-450 reductase, directly demonstrate the formation of superoxide anions. It is concluded that oxycytochrome P-450 may decompose to generate hydrogen peroxide.


Comparison of the peroxidatic activity of cytochrome P-450 with other hemoproteins and model compounds. The H2O2 dependent catalysis of cytochrome P-450 was compared with the catalytic mechanism of horse radish peroxidase, methemoglobin and iron protoporphyrin complexes. A relatively stable intermediate being comparable to compound I of horse radish peroxidase is formed in the case of iron porphyrin complexes, methemoglobin and probably cytochrome P-450. In the case of peroxidase compound II is the more stable intermediate. This could be the reason for the different catalytic properties of peroxidase on the one hand and iron porphyrin complexes, methemoglobin and cytochrome P-450 on the other hand.


Serum levels of somatomedin A and growth during long-term treatment of patients with pituitary dwarfism with human growth hormone. Eighteen patients with pituitary dwarfism were treated for 1 7/12 to 6 years with human growth hormone (hGH) at a dose of 0.19--0.62 unit (U) per kg of body weight per week. The mean increment in height was 2.0 +/- 0.4 and 8.8 +/- 0.5 cm/year before and the first year after treatment of hGH, respectively. A significant positive correlation was observed between serum levels of somatomedin A and growth rate, espcially in children with bone age below 10 and a duration of treatment of less than one year (r = 0.66, P less than 0.005). Long-term treatment with hGH was accompanied by a decreasing response. However, the serum levels of somatomedin A did not decrease significantly. Therefore, decreased growth increment in these situations was not due to decreased serum levels of somatomedin A.


Sex difference in the induction of lactogenic receptors in rat livers. The relationship between serum levels of growth hormone (rGH), prolactin (rPRL), luteinizing hormone (rLH), follicle-stimulating hormone (rFSH), corticosterone, oestrogen, (oestradiol-17 beta) and testosterone and the hepatic binding sites specific to [125I]human-prolactin (h-PRL) were investigated in normal rats, in rats bearing the GH- and PRL-secreting tumour (GH3), and in rats 14 days after tumour removal. The presence of GH3 tumour elevated serum levels of rGH and rPRL and concomitantly increased the hepatic binding of [125I]h-PRL; the male rats had a greater increase than the female rats. The increased binding was due to an increase in the specific membrane binding sites, whereas the affinity constant (Ka) was not changed. In both male and female rats, there was a significant positive correlation between ser-m rGH levels (P less than 0.001) or serum rPRL (P less than 0.02) and specific binding of [125I]h-PRL in female rats only (P less than 0.02). In male rats, there was a significant negative correlation between serum testosterone levels and specific bindings of [125I]h-PRL (P less than 0.05). These results suggest that rGH and rPRL regulates the hepatic h-PRL receptors in female rats, and testosterone predominantly inhibits the induction of the hepatic lactogenic receptors in male rats.


Hormonal effects on rat mammary gland in vitro. The mammogenic and lactogenic effect of various hormones on rat mammary gland has been studied in vitro using an organ culture technique. The effects have been assessed mainly by light microscopy. The study has been confined mainly to the effects and interactions of the lactogenic protein hormones, corticosteroids and progesterone. It establishes the importance of the presence of insulin and serum and of the type of serum for the hormonal effects in vitro. These effects have been studied mainly on the explants obtained from 13 days pregnant rats. In addition explants from non-pregnant rats and at various stages of pregnancy and lactation were studied.


Character of the gastric and duodenal electrical activity after blocking of the adrenoreactive structures. Experiments are carried out on six dogs with chronically implanted electrodes in the gastric wall and in the duodenum. The electrical activity of the stomach (EGMG) and of the duodenum (EEG) is recorded. The myoelectrical complex of the stomach consists of four phases. The first phase records only slow potentials and lasts 40--60 min. The appearance of spike activity in the duodenum is always preceded by EGMG activation. Intravenous injection of the specific blocker of the alpha-adrenergic structures (phentolamine -- 1 mg/kg) leads to the appearance of spike activity in EGMG and EEG. 50--60 per cent of the slow waves being accompanied by spike potentials. At the same time the frequency of the slow potentials from EGMG is also decreased. Application of beta-adrenoblocker (propranolol -- 2 mg/kg) does not change subatantially EGMG and EEG. Simultaneous administration of alpha- and beta-adrenoblockers prolongs spiking time in the stomach and duodenum (spike potentials are recorded for 120--180 min, duration in the background 40--50 min). The influence of the interactions of cholinergic and adrenergic structures on the character of EGMG and EEG is discussed.


The neural control of the serotonin content in mammalian enterochromaffin cells. Serotonin (5-HT) in the mammalian gut is mainly stored in enterochromaffin cells (EC). Vagal or splanchnic nerve stimulation respectively increase portal blood levels of 5-HT and decrease 5-HT content in EC, suggesting a neural control of these cells. Previous studies have also suggested that the vagal effect is mediated via vagal adrenergic nerve fibers, since this effect could be blocked by sympathectomy, removal of the superior cervical sympathetic ganglia or by the administration of a beta-adrenoceptor blocking agent, d, 1-propranolol. In the present study the control of the 5-HT content in EC was studied by a cytofluorimetric method in specimens from rat duodenal mucosa after drug incubations and transmural field stimulation (TFS) in vitro: Incubations in a Krebs' solution containing adrenaline (A), noradrenaline (NA) or isoprenaline (IP) (10(-9)--10(-5) M) caused a decrease of the 5-HT content in EC. For NA and A this effect was concentration-dependent. The effect of A was antagonized by d, 1-propranolol but not by d-propranolol, metoprolol, phentolamine or phenoxybenzamine. This indicates that the effects observed were mediated via a true beta-adrenoceptor mechanism. Acetylcholine (ACh), in the presence of eserine, also decreased the 5-HT content in EC, but was less potent than the adrenergic substances. TFS decreased the 5-HT content of EC to about 75% of unstimulated control levels. The presence of tetrodotoxin or d, 1-propranolol in the stimulation bath antagonized the effect of TFS. In specimens from rats pretreated with 6-OH-dopamine, TFS had no effect. Thus, TFS appears to induce a release of 5-HT from EC mainly by activation of adrenergic nerves. The ultrastructural relationship between nerve terminals and EC was studied in the guinea-pig duodenum. Animals were pretreated with 5-OH-dopamine in order to visualize adrenergic nerves. Near the base of most of the EC examined, at least 3 different types of nerve terminals were observed with the appearance of adrenergic, cholinergic and peptidergic terminals. In addition, dendrite-like processes were demonstrated. No true synapse on EC was found, but the minimal distance between the nerve terminals and EC was 150-250 nm, well within the limits of the "autonomic gap", suggesting a functional neural influence on these cells. The importance of the demonstrated beta-adrenergic control of EC was further studied in vivo. Efferent vagal nerve stimulation in the cat caused increased 5-HT levels in the portal plasma to more than twice the normal values after 15 min stimulation. Administration of the beta-adrenoceptor antagonist d, 1-propranolol in various doses did not abolish, but significantly reduced the response, particularly during the final part of the stimulation period. These results not only confirm a beta-adrenoceptor mediated release of 5-HT, but also suggest that other mechanisms are involved in the total response to vagal nerve stimulation.


Lipoprotein abnormalities in cholestasis. II. Isolation, characterization and clinical evaluation of an additional cholestatic lipoprotein (slow-migrating HDL). An additional cholestatic lipoprotein with a slower mobility than the usual alpha-lipoprotein on polyacrylamide-gel disc-electrophoresis was found in the serum of patients with cholestasis. This abnormal lipoprotein was referred to as Slwo-migrating HDL (HDL-S), because it was mostly recovered in the high density lipoprotein (HDL) fraction after preparative ultracentrifugation. HDL-S was precipitated by dextran sulfate and Mg++ but did not react with either concanavalin A or anti-beta-lipoprotein serum. The main apoprotein of HDL-S was Apo A-I, and a trace of Apo E was also present. HDL-S was relatively enriched in free cholesterol and triglycerides and had a density in the range of 1.063 to 1.083. The appearance of HDL-S in serum or plasma was closely associated with chronic mild intrahepatic cholestasis, particularly as in primary biliary cirrhosis and related conditions.


Decreased disappearance rate of 1-(2-tetrahydrofuryl)-5-fluorouracil (FT-207) from the blood and its unresponsiveness to tocophreyl nicotinate and indomethacin in patients with primary hepatocellular carcinoma. The disappearance rates(K) of FT-207 from the blood in patients with primary hepatoma and advanced cirrhosis of the liver were significantly lower than those in control patients with cancer but normal liver function. Pretreatment with tocopheryl nicotinate and indomethacin increased the K values in the control subjects, but was without effect on the K values in patients with primary hepatoma.


Recent concepts in the conservative treatment of intracranial tumours in children. The role of radiotherapy for certain intracranial tumours, given either postoperatively or as the sole treatment in inoperable cases, is discussed, principally in relation to medulloblastoma, ependymoma, cerebral astrocytoma, brain-stem glioma and craniopharyngioma. Methods of increasing the response of gliomas to irradiation are mentioned and reference is made to the possible value of chemotherapy and of immunotherapy as adjuvants against such tumours. Treatment results are reported and consideration given to the quality of long-term survival.


CSF levels of cyclic nucleotides in primary intracranial neoplasms: a preliminary report. Recent evidence indicates that cyclic nucleotides are of importance for general and neurosurgical oncology, especially with respect to the contact-inhibition mechanisms and tumour cell growth. This preliminary report deals with the CSF levels of c-AMP and c-GMP in primary neoplasms in children and to problems related to the blood-brain barrier. Some cases of medulloblastoma were studied as well as a few cases of brain stem glioma and cystic astrocytoma. The importance of some rather unusual findings seems undebatable, i.e., the marked increase in c-GMP values usually observed in medulloblastoma and the decrease of c-AMP, that is fairly common in all malignant neoplasms. The main changes in the c-AMP/c-GMP ratio are also discussed.


Cerebral mixed tumour: osteo-condrosarcoma--glioblastoma multiforme. A 49-year-old woman had a left frontal expanding mass with calcified areas. A craniotomy and resection of the mass were performed. Histological sections showed a mixed tumour: osteochondrosarcoma-glioblastoma multiforme. The absence of previous irradiation and its location, deep in the frontal lobe and apparently unattached to the meninges, add to the rarity of this association. A brief discussion concerning the presence of osseous or cartilaginous areas in brain tumours and the problem of cerebral mixed tumours follows the description of the case.


Endothelial cells and components of the kallikrein-kinin system. Endothelial cells are a major source of kininase enzymes including kininase II. Kininase II is situated along the plasma membrane, not as an ecto-enzyme but as an enzyme synthesized by the endothelial cells themselves. However, it is likely that endothelial cells do more than degrade kinins. These cells are contractile and may possess kinin receptors; a possibility supported by the fact that kinins stimulate endothelial cells to form and release prostaglandin-related substances. In addition, we have found that endothelial cells in culture are reactive with antibodies to alpha 2-macroglobulin. Endothelial cells can hydrolyze [3H]Pro-Phe-Arg-anilide, a kallikrein substrate, but the reaction is not inhibited by soya bean trypsin inhibitor (SBTI) or Trasylol. Possibly kallikrein or a related trypsin-like enzyme is bound to alpha 2-macroglobulin and is not free to react with the inhibitors. Thus, endothelial cells can bind and inhibit kallikrein-like enzymes, degrade kinins and respond to kinin stimulation.


The effect of human granulocyte proteinases on kininogens. From the results obtained in preliminary in vitro experiments we may conclude that the unspecific breakdown of kininogens by neutral and acidic granulocytic proteinases may occur also in the organism during pathological conditions. Inhibitor application may serve as a valuable tool to prevent unspecific proteolytic degradation and thus elimination of kininogens and other clotting or complement factors from their biological function. Furthermore, specific cleavage products of the kininogens produced by granulocytic enzymes may be useful in the diagnosis of a beginning septicemia as well as in elucidation of the primary structure of the human kininogens.


Comparative clinical study of FOY and Trasylol in acute pancreatitis. In order to evaluate the efficacy and safety of FOY injectable ([ethyl-4-(6-guanidinohexanoyloxy) benzoate] methane sulfonate) on acute pancreatitis, a comparative clinical study was carried out using Trasylol as the control at 38 hospitals in Japan. Favourable results were obtained in 60 (71%) out of 84 patients in the FOY group and 29 (44%) out of 66 patients in the Trasylol group. The results showed that these both drugs were effective and the statistical analysis revealed considerable difference (X2 = 10.464, p less than 0.005) between the two groups in this condition of clinical trial. In addition, a double blind trial was carried out at 4 hospitals using FOY-305 ([n.n-dimethylcarbamoylmethy 4-(4-guanidinobenzoyloxy)-phenylacetate] methane sulfonate) oral capsule and inactive placebo. Favourable results were obtained in 18 (69%) out of 26 patients in the FOY-305 group and 8 (32%) out of 25 patients in the inactive placebo group, indicating a significant difference between the two groups (X2 = 8.930, p less than 0.01). The results of the present study suggest that synthetic protease inhibitor, FOY or FOY-305, is beneficial in the treatment of acute pancreatitis.


Inhibitory effects of gabexate mesilate (FOY) on experimental DIC. The inhibitory effects of a newly synthesized protease inhibitor, Gabexate mesilate (FOY), on experimental disseminated intravascular coagulation were studied as compared with those of aprotinin or heparin. Thrombin, tissue thromboplastin, and endotoxin were used as DIC trigger substances. As parameters on DIC, platelet counts, white blood cell counts, neutrophilic leukocyte counts, fibrinogen, fibrin degradation products, platelet retention, platelet aggregation, prothrombin time, partial thromboplastin time were served. The drug efficacy in each parameter were expressed by the score system and analyzed statistically. The results were summarized as follows; (1) In thrombin-induced DIC, FOY was apparently superior to the other drugs (p less than 0.05). (2) In thromboplastin-induced DIC, heparin was slightly more effective than FOY or aprotinin. (3) In endotoxin infusion, there were no significant differences among them. In conclusion, the results of the present study suggest that FOY was more effective than heparin or aprotinin on experimental DIC.


Effect of hypophysectomy upon renal kallikrein-kinin system in rats. It is confirmed that urinary kallikrein is significantly and persistently decreased in totally hypophysectomized rats. In addition, kallikrein activity in the kidneys in these animals is significantly reduced (p less than .001) as compared to partially hypophysectomized and sham-operated rats. The administration of 125 mU of ACTH twice a day for 2 months, did not modify urinary kallikrein excretion. Negative effects were also obtained in subsequent periods, by injection of 10 microgram corticosterone and 10 microgram aldosterone, twice a day for 9 and 6 days, respectively. The administration of rat pituitary anterior lobe homogenates, freshly prepared, increased significantly urinary kallikrein level in hypophysectomized rats.


Purification and properties of angiotensin I-converting enzyme in human lung and its role on the metabolism of vasoactive peptides in pulmonary circulation. Purification of angiotensin I-converting enzyme from human lung and characteristics of the enzyme was studied. Experimental pneumonitis was produced in rabbits and the change of the activity of angiotensin I-converting enzyme was studied in purpose to clarify the role of this enzyme in the metabolism of vasoactive peptides in the lung. Purification was performed using trypsin treatment, acid treatment, DE52-cellulose column chromatography, hydroxyapatite chromatography and Sephadex G-200 gel filtration. The enzyme after final step showed a single band on disc gel electrophoresis. Experimental pneumonitis was produced by injection of Complete Freund's adjuvant (acute pneumonitis) and of N-nitroso-N-methylurethane (chronic pneumonitis). In acute experiment, angiotensin I-converting enzyme activity in pulmonary tissue and in plasma was significantly decreased. In perfusion experiment, conversion of angiotensin I to angiotensin II and inactivation of bradykinin were also significantly decreased. In case of decreased activity of angiotensin I-converting enzyme in the lung, less angiotensin II will be released into systemic circulation and bradykinin will pass through the pulmonary circulation into systemic circulation, thus this may result in the decrease of systemic blood pressure.


Relationship of human urinary and plasma kinins to sodium-retaining steroids and plasma renin activity. In these studies we have shown that: 1. urinary kallikrein is an index of sodium-retaining steroid activity and may participate in the anti-natriuretic and kaliuretic effects of these hormones; 2. urinary kinin excretion is independent of changes in urinary kallikrein excretion; 3. levels of plasma bradykinin are highly correlated with levels of plasma renin activity probably because both respond to changes in extracellular fluid volume; and, 4. plasma bradykinin may act to antagonize angiotensin II to maintain normal blood pressure in hyper-reninemic states.


Mammalian inhibitors of angiotensin converting enzyme (kininase II). Urines and sera (human, guinea pig and rat) contain low molecular weight inhibitors of angiotensin converting enzyme (ACE). The urines contain ACE, but the enzyme is scarcely measurable without prior ultrafiltration or dialysis. The activity increases strikingly through three ultrafiltration steps using a membrane with a 10,000 MW retention limit. As implied, the ultrafiltrates contain inhibitory activity and can prevent the hydrolysis of [3H]benzoyl-Gly-His-Leu by ACE from any source, including lung and serum. Human urinary ultrafiltrate contains at least three inhibitors separable on Bio-Gel P-2. The inhibitors are acidic and can be partially purified on Bio-Rex 70 developed with an acetic acid gradient. The smallest of the inhibitors can be purified to apparent homogeneity by partition chromatography (sephadex G-25; butanol, acetic acid, H2O; 4:1:5). The excretion of inhibitory activity varies in response to dietary salt: Activity is low when rats are maintained on a high NaCl diet and is high (3 x's control) on a low NaCl diet. Thus, the activity of ACE may be modulated in vivo by naturally-occurring enzyme inhibitors. Whether some hypertensive patients are deficient in ACE inhibitory activity remains to be determined.


A postsynaptic inhibitory histamine H2-receptor in the mouse isolated vas deferens. The effect of histamine on adrenergic neurotransmission in the mouse isolated vas deferens preparation was investigated. Concentrations of histamine ranging from 0.2 to 650 microM depressed, in a dose-related manner, not only the contractile response elicited by field stimulation but also the response caused by the addition of exogenous noradrenaline and acetylcholine. However, the release of [3H]-NA evoked by field stimulation or by high K+ remained unchanged in the presence of these concentrations of histamine. The inhibitory effect of histamine on the contractile responses caused by various stimuli was reduced or completely antagonized by cimetidine, a histamine H2-receptor antagonist but not by mepyramine, a conventional antihistamine. The inhibitory effect of histamine was found to be inversely proportional to both the Ca2+ concentration in the bathing medium and to the frequency of field stimulation. Further, the inhibitory effect of histamine was markedly reduced when Mg2+ was omitted from the bathing medium. It is concluded that the mouse vas deferens preparation contains a post-junctional inhibitory H2-receptor. The stimulation of H2-receptors by histamine inhibits the contractile response of the vas deferens, possibly by decreasing the availability of Ca2+ required for contraction by depressing the influx of Ca2+.


Action of mannitol in various immunological experimental models. The authors describe the inhibiting action of mannitol after repeated administration of low subcutaneous doses in a number of experimental immunological models. For example, in the rat it produces a reduction of the secondary arthritis of Freund's adjuvant polyarthritis and also of the pleurisy due to Bordetella pertussis hypersensitivity. In the mouse it reduces the reaction of delayed hypersensitivity to sheep red cells. Its action is also marked against ovalbumin-induced active skin anaphylaxis in the albino guinea-pig and on IgE synthesis in the rat. Moreover, after several injections it produces a reduction of carbon phagocytosis in the mouse. At the doses at which the effect appeared, no action could be found on various models of acute non-immune inflammation, diuresis, blood pressure, hematocrit and protein and plasma sodium levels.


Solid and mucinous varieties of so-called mammary carcinoid tumors. Three neoplasms that had histologic features reminiscent of carcinoid tumors of other sites were encountered in a review of 3,300 examples of invasive mammary cancer in women (.09%). One of these showed cytoplasmic argyrophilia. This, as well as the two putative carcinoids, lacked argentaffinity. Attention is directed to the occurence of variable numbers of argyrophilic cells in eight of 19 so-called mucinous cancers of the breast studied. Further, neurosecretory-type granules were observed in cells of all four mucinous cancers suitably prepared for electron microscopic examination. The possible reasons for the lack of universal argyrophilic reactions in these lesions is discussed. It is concluded that there may be two types of mammary carcinoid tumors, the solid and mucinous varieties. No patient who had the latter type had experienced treatment failure after five years of observation. Various numbers of ductal epithelial cells in four of 45 examples of banal fibrocystic disease showed cytoplasmic argyrophilia, and neurosecretory-type granules were found in two of eight examples suitably prepared for electron microscopic examination. Whether this demonstration establishes the existence of precursor elements for the development of the carcinoid tumors is at present uncertain.


Storage and transport of cultures for Herpes simplex virus, type 2. Tryptic soy broth and brain-heart infusion broth maintained herpes simplex virus within +/- 1 serial dilution of the original titer in all of 51 samples held for one to three days at 4 C. In contrast, holding temperatures of -20 C or 22 C resulted in losses of titer of 10(2) or more. Holding periods as long as five days prior to inoculation did not delay the appearance of the cytopathic effect following inoculation. Unmodified tryptic soy broth or brain-heart infusion broth and holding periods as long as three days at 4 C prior to arrival in the laboratory give as satisfactory results as do traditional viral culture media and prompt inoculation of the specimen.


Transfer of measles, mumps, and rubella antibodies from mother to infant. Its effect on measles, mumps, and rubella immunization. Sera from 42 mother-infant pairs were examined to determine the effect of passively acquired enhanced neutralizing (ENt) antibody on immunization. The ENt antibodies to measles, mumps, and rubella were greater in term newborns than in their mothers, with mean ratio of 1.8:1, 1.3:1, and 1.2:1, respectively. In 21% to 25% of the children, these antibodies persisted until 12 months of age. When immunized with trivalent measles-mumps-rubella vaccine, children who had persisting ENt measles and rubella titers had significantly lower mean antibody responses than children without detectable antibodies to the two viruses. Persisting ENt mumps antibodies did not affect the postimmunization titers. Seroconversion rates to any of the three viruses were not different in children with or without preexisting ENt antibody.


Vitamin D metabolism and calcium absorption. Vitamin D along with parathyroid hormone (PTH) and calcitonin (CT) are the three principal effectors of calcium and phosphorus homeostasis. The secosteroid, vitamin D3, is subject to metabolic conversion to its biologically active form(s) 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] and 24,25-dihydroxyvitamin D3 [24,25(OH)2D3] prior to initiation of its physiologic responses in the intestine and skeletal system. The production of 1,25(OH)2D3 is stringently regulated by a variety of endocrine signals including PTH as well as the "calcium needs" of the organism. At the target intestine, 1,25-(OH)2D3 stimulates the intestinal absorption of calcium via a mechanism analogous to that of other steroid hormones. Definitive biochemical evidence exists supporting the existence in the intestine of a highly specific protein receptor for 1,25(OH)2D3. After formation of the steroid-receptor complex, it migrates to the nucleus of the cell and stimulates messenger-RNA synthesis for proteins (including a calcium-binding protein) which are necessary for the generation of the biologic response. Current efforts to biochemically characterize vitamin D-mediated intestinal calcium transport include efforts to understand the role of calcium-binding protein in this process, as well as to identify other protein components present either in the brush border or basal lateral membranes.


Pectoralis major muscle defect and Poland complex. Pectoralis major muscle defect (PMD) was diagnosed in 27 infants from a series of 599,109 live births in South America (1/22,189). In all 27 cases the PMD was unilateral, mainly affecting the right side (20/27), and there were more male (19/27) than female cases. No familial cases and no parental consanguinity were recorded. A positive correlation was observed between PMD and sex hormone intake and vaginal bleeding in the first trimester of pregnancy. In 12 (1/49,925) of the 27 PMD cases hypoplasia and/or syndactyly of the ipsilateral hand was also diagnosed. The index-middle interdigital space was affected in all 11 cases with symbrachydactyly. Additional congenital anomalies were observed in 4/27 cases, and they were: hemangiomas, hypospadias, and clubfeet. Poland complex (12 cases), isolated PMD (15 cases), and isolated symbrachydactyly (18 cases), showed a similar pattern for symmetry, sidedness, syndactyly type, and sex ratio.


Bilateral cytomegalovirus panuveitis after high-dose corticosteroid therapy. A 30-year-old previously healthy woman who recieved immunosuppressive doses of corticosteroids developed an unusual culture-proved cytomegalovirus panuveitis bilaterally. The atypical clinical course was marked by bilateral exudative detachements with minimal evidence of retinitis. Any patient receiving blood transfusions and immunosuppressive doses of corticosteroids is potentially at risk for developing ocular cytomegalovirus infection, and the clinical presentation of the ocular infection may be atypical.


Developments in the identification of human skeletal material (1968-1978). A review of publications over the last ten years of methods in the field of physical anthropology for determining age, sex, race and stature of human skeletal material is presented. Comparisons are made with the types of papers published in the previous ten years (1958-1968) in six categories: (1) Visual examination of bones (2) Anthropometric measurements of bones (3) Anthropometric measurements with subsequent use of statistics in the form of discriminate function analyses (4) Time and sequence of eruption of the teeth (5) X-ray examination of the internal structure of bone sections (6) Microscopic examination of the internal structure of bone.


Urinary free cortisol excretion in patients with metastatic cancer. Many patients with metastatic carcinoma have an increase in urinary excretion of 17-hydroxycorticosteroids (17-OHCS), plasma concentrations of 17-OHCS, plasma total cortisol, plasma nonprotein-bound cortisol, or cortisol production rate. A small portion of circulating cortisol is excreted in the urine as unaltered or free cortisol. This urinary free cortisol, which is believed to reflect the average unbound plasma cortisol circulating during the day, is an excellent measure of adrenal cortical function. In this study, we compared the urinary free cortisol of 35 patients with metastastic cancer (10 small cell carcinomas of the lung, 8 other types of carcinoma of the lung, 8 breast carcinomas, and a miscellaneous carcinomas) with a group of 15 healthy age-matched volunteers. There were no significant changes in urinary free cortisol excretion in the patients with cancer.


Effect of ACTH on the aldosterone response to potassium in sheep with adrenal transplants. The importance of physiological ACTH stimulation in maintaining the response of aldosterone secretion to potassium was studied in 5 conscious sheep with cervical adrenal autotransplants. Endogenous ACTH secretion was suppressed by dexamethasone. Constant local infusions of potassium that raised adrenal venous plasma by 2 mmol/l increased aldosterone secretion from 3 +/- 1 ng/min (mean +/- SE) to levels of 50 +/- 13 ng/min at 30 min, after which secretion fell to 14 +/- 6 ng/min at 230 min. Addition of submaximal ACTH (0.04 - 0.10 mU/min) to potassium infusions produced similar responses at 0--130 min, but aldosterone secretion increased at 130--190 min and was more sustained (40 +/- 9 ng/min at 230 min; P less than 0.05). Infusions of submaximal ACTH alone did not significantly increase aldosterone secretion above basal levels. Infusions of maximal ACTH (16.6 mU/min) produced higher aldosterone secretion in the group given ACTH replacement. These results show that the aldosterone response to potassium is phasic and poorly sustained in the absence of ACTH. Responsiveness can be restored by doses of ACTH insufficient alone to stimulate aldosterone secretion.


Physiological basis for absorptive and renal hypercalciurias. Idiopathic hypercalciuria constitutes two major variants-absorptive hypercalciuria, characterized by a primary intestinal hyperabsorption of calcium, and renal hypercalciuria, in which renal tubular reabsorption of calcium is primarily impaired. The two forms of hypercalciuria may be distinguished from each other, since a) parathyroid function is stimualted in renal hypercalciuria, but normal or suppressed in absorptive hypercalciuria, b) the renal leak of calcium is present in renal hypercalciuria, but not in absorptive hypercalciuria, c) intestinal calcium absorption is probably increased primarily in absorptive hypercalciuria, and secondarily in renal hypercalciuria (from parathyroid hormone excess), d) the increased calcium absorption in renal hypercalciuria probably results from the parathyroid hormone-dependent stimulation of 1,25-dihydroxyvitamin D synthesis, whereas that in absorptive hypercalciuria may be vitamin D-independent, e) the response of the two conditions to certain treatments is unique, and f) the sequelae of parathyroid hormone excess, such as low bone density and negative calcium balance, may be present in renal hypercalciuria, but not in absorptive hypercalciuria. These findings provide a physiological basis for the consideration of absorptive and renal hypercalciurias as distinct and separate entities.


Indirect evidence for enhancement of Na-K-ATPase activity with stimulation of rectal gland secretion. The relation between active chloride secretion and Na-K-ATPase activity was studied in the isolated perfused rectal gland of Squalus acanthias stimulated by cAMP and theophylline. Ouabain inhibited both secretion and oxygen uptake in stimulated glands. Stimulation of chloride secretion by cAMP and theophylline resulted in a six- to sevenfold rise in ouabain-inhibitable oxygen consumption. Intracellular sodium concentration decreased while intracellular potassium rose. These findings suggest activation of Na-K-ATPase. On the other hand, the activity of Na-K-ATPase in whole homogenates of rectal gland tissue was not increased after cAMP stimulation of the perfused gland, nor was ouabain binding increased. Calculated intracellular chloride concentration fell after secretion was stimulated in a way consistent with an action of cAMP to increase luminal permeability to chloride.


Mixed neuromuscular block. A re-assessment using train-of-four stimulation. Suxamethonium was administered to patients during recovery from non-depolarising muscle relaxation. The effect of suxamethonium varied depending upon the degree of recovery from the non-depolarising block when it was administered. Early in recovery suxamethonium produced reversal of the non-depolarising block without paralysis whereas when administered later, initial recovery of the non-depolarising block was followed by paralysis. If administered after 50% recovery the predominant effect was paralysis without initial recovery. Assessment of neuromuscular blockade with train-of-four stimulation showed that the paralysis produced by suxamethonium under these circumstances, had depolarising and non-depolarising features.


Pathogenesis of swine vesicular disease in pigs. Pigs exposed to swine vesicular disease virus developed vesicular lesions by postinoculation day 2. Lesions first appeared on the coronary band and then on the dewclaw, tongue, snout, lips, and bulbs of the heels. The onset of viremia coincided with febrile response and the appearance of vesicles. Virus was isolated from the nasal discharge, esophageal-pharyngeal fluid, and feces as early as postinoculation day 1. Greater amounts of virus were isolated from samples collected during the first week of infection, and lesser amounts from samples collected during the second week. The appearance and the distribution of specific fluorescence in various tissues indicated that during the development of swine vesicular disease virus infection, the epithelial tissues were initially involved, followed by a generalized infection of lymph tissues, and subsequently, a primary viremia. Seroconversion was detectable as early as postinoculation day 4. A mild nonsuppurative meningoencephalomyelitis throughout the CNS was observed in both inoculated and contact-exposed pigs. The olfactory bulbs were most severely and were frequently affected, particularly in contact pigs. The most severe brain lesions were found in pigs 3 to 4 days after the onset of viremia; contact pigs showed more severe brain lesions than inoculated pigs. Microscopic changes were also found in the coronary band, snout, tongue, and heart.


Reversal of pancuronium with edrophonium. Edrophonium was administered as a bolus of 2, 10 or 50 mg (0.03, 0.14 and 0.71 mg/kg) to antagonize neuromuscular block after pancuronium in man. Neuromuscular transmission was measured using "train-of-four" stimulation. Recovery started within 1 min, was maximum at 2 min and was then followed by a plateau of slow continued improvement. The degree of recovery increased with larger doses of edrophonium and also was related to the total amount of pancuronium administered although reversal was attempted at the same degree of spontaneous recovery. There was no evidence of recurarisation or depolarising block.


Transmissible gastroenteritis in neonatal dogs: experimental intestinal infection with transmissible gastroenteritis virus. Fourteen neonatal dogs (4 through 11 days of age) were exposed orally to the Purdue strain of transmissible gastroenteritis (TGE) virus, and six dogs of similar age were noninoculated controls. Clinical signs of enteric disease did not develop. Both exposed and control dogs had normal fecal passages and appetite throughout the experiment. Jejunal epithelium from dogs euthanatized at 12, 24, 48, and 96 hours and at 10 days after exposure did not exhibit morphologic alterations detectable by light microscopy. Electron microscopic examination indicated that jejunal epithelial cells contained TGE viral particles as early as 12 hours after dogs were exposed. There were no apparent morphologic alterations or signs of desquamation of virus-infected cells, however. Results of pig transmission studies indicated that viable TGE virus was in jejunal tissue of the dogs as early as 12 hours and as late as 10 days after exposure to the virus.


Three serologic types of adenovirus infections of owl monkeys. From a colony of 131 owl monkeys, adenovirus was isolated 68 times from 433 oropharyngeal and rectal swab sample pairs collected over a 2-year period. A total of 63 of 68 adenovirus isolates were grouped by neutralization into 1 of 3 types designated owl monkey adenovirus types (OMAV Ty) I, II, and III. The OMAV Ty I and Ty II were identified by neutralization with antiserum to squirrel monkey adenovirus type I and adenovirus SV-11, respectively. The OMAV Ty III was partially neutralized by antiserum to OMAV Ty II. The OMAV Ty I was isolated from 8 of 30 newly imported owl monkeys within 3 weeks of arrival. A total of 13 of 26 owl monkeys had a fourfold or greater rise in serum-neutralization (SN) titer with 25 of 27 developing SN titers of greater than or equal to 1:80. A total of 40 of 87 other owl monkeys in the colony had SN titers of greater than or equal to 1:5. The OMAV Ty II was isolated from 51 (12%) of 433 swab samples pairs. Virus was isolated from 26 owl monkeys with 13 having persistent infections. Viral isolations spanned a 10- to 17-month period for five of these owl monkeys. In 114 owl monkeys, 101 (86%) had SN titers to OMAV Ty II greater than or equal to 1:20. The OMAV Ty III was isolated from three owl monkeys. Of 114 owl monkeys, 93 (82%) had SN titers to OMAV Ty III of greater than or equal to 1:20. Although the distribution of SN titers were similar for OMAV Ty II and Ty III, 42 (37%) of 114 owl monkeys had eightfold or greater differences in SN titers between OMAV Ty II and Ty III.


Comparative pathogenesis studies with oncogenic and nononcogenic Marek's disease viruses and turkey herpesvirus. An apparently nononcogenic Marek's disease virus (SB-1) and turkey herpesvirus could be readily isolated from spleen, bursa of Fabricius, thymus, and peripheral blood lymphocytes of chickens beginning 4 to 6 days after inoculation, but unlike infections with two isolates of oncogenic Marek's disease virus (JM-10 and CU-2), virus replication in these cells was rare, and necrosis in the organs was essentially absent. Splenic enlargement was observed regularly during the first 4 to 11 days after inoculation, and Marek's disease tumor-associated surface antigen was observed on splenic and other lymphocytes in the four viral inoculation groups. Cellular cytotoxicity of splenic lymphocytes was demonstrated in vitro with cultured Marek's disease tumor cells (MSB-1 lymphoblastoid cell line) as the target in a chromium-release assay. The four viral infections induced sensitized lymphocytes.


Inhibition and enhancement of avian adenovirus plaque production by heavy and light avian adenovirus-associated viral particles. The effect of heavy and light avian adenovirus-associated viral (A-AV) particles on the replication of several adenovirus serotypes was studied in chicken embryo kidney cells. There was a significant decrease (P less than 0.05) in the number and size of adenovirus-induced plaques at A-AV multiplicities of infection greater than 40. Enhancement of plaque production was observed when A-AV multiplicities of infection were 1 to 40. There was a significant increase in the number and size of infective centers. Analysis of cellular yields indicated an increase in the number of adenoviruses produced per cell. Heavy A-AV particles of buoyant density 1.42 g/cm3 in CsCl were found to enhance plaque production more than light particles (1.38 g/cm3). Conversely, light particles showed greater inhibition of plaque production. Adenovirus serotypes varied in their response to enhancement or inhibition by A-AV particles of different density.


Effects of Sendai virus infection on function of cultured mouse alveolar macrophages. Cultured mouse alveolar macrophages supported the growth of Sendai virus (murine parainfluenza I virus), as measured by both a 10-fold increase in extracellular virus titers and development of viral antigens on most of the cells. Synthesis of virus continued for at least 1 month without cytopathic effects. Macrophage phagocytic activity for Candida, Staphylococcus epidermidis, and opsonized erythrocytes remained unaffected by the infection, and the ability of the cells to kill S. epidermidis and S. Aureus was also unchanged. The defects in alveolar macrophage function observed in Sendai-infected mouse lungs probably are not due to a direct effect of the virus on macrophage function.


Peripheral vascular disease following radiation therapy. Radiation therapy is effective for the treatment of many malignant diseases. This fact is well documented. The existence of complications in any treatment form does not obviate its usefulness. An awareness of possible complications, however, makes our treatment program more realistic and causes us to be more aware of the patient's complaints following therapy. One of our patients complained of claudication for seven years before an arteriogram was done. How many swollen, painful arms after mastectomy and irradiation have been passed off as due to lymphedema or a complaining patient? Exertional chest pain in a young person after mediastinal irradiation should not be overlooked. These symptomatic lesions are amenable to surgical correction. Great care must be spent when incising irradiated skin areas. The use of Silastic sheeting may be useful in extreme cases of radiation fibrosis involving the ureter. While no human studies are available for documentation, based on animal studies, it may be useful to decrease lipidemic situations through dietary prudence during radiation therapy. Finally, we are reminded again that many of our therapeutic triumphs are tarnished with serious side effects.


Changing patterns of breast cancer.

Changing patterns of breast cancer. Changes in treatment of breast cancer are high-lighted as follows: 1) Outpatient breast biopsy offers advantages in planning treatment and also saves money, 2) Mammograms are increasing the numbers of breast tumors found early, 3) For many reasons modified mastectomy is finding increasing use, 4) Breast cancer appears to be multifocal at least one-half of the time. Multifocal breast cancer is more aggressive, 5) Bilateral breast cancer appears to have a very favorable prognosis, especially when non-simultaneous.


Plasma and urine cyclic guanosine 3':5'-monophosphate in disseminated cancer. Plasma and 24-h urinary adenosine 3':5'-monophosphate (cyclic AMP) and guanosine 3':5'-monophosphate (cyclic GMP) were measured by radioimmunoassay in 12 normal subjects, 33 patients with six types of non-neoplastic disease (cholelithiasis, peptic ulcer, coronary heart disease, hypertension, regional ileitis, and cirrhosis), and 34 patients with five types of disseminated neoplastic disease (acute myelocytic leukemia; Hodgkin's disease; and metastatic cancer of the lung, colon, and breast). In patients with non-neoplastic disease, cyclic nucleotide values in plasma and urine did not differ significantly (P greater than 0.05) from those in normal subjects. In patients with disseminated cancer, cyclic AMP values in plasma and urine likewise did not differ significantly from those in normal subjects. Plasma cyclic GMP, in contrast, was significantly elevated in all five types of cancer patients, and urinary cyclic GMP was significantly elevated (five times the normal mean) in patients with acute myelogenous leukemia and Hodgkin's disease.


Small cell undifferentiated carcinoma of the larynx. A patient with a small cell, undifferentiated carcinoma of the larynx is described. The tumor appeared to arise in close association with subglottic minor salivary glands. Ultrastructural study of the tumor cells showed neurosecretory granules similar to those seen in normal Kulchitsky cells and pulmonary oat cell carcinomas. Bronchogenic oat cell carcinoma and small cell undifferentiated carcinoma of minor salivary gland tissue probably originate from the same cell type. The aggressive behavior of this tumor suggests the need for early systemic treatment including combination chemotherpay.


Cyclic adenosine 3', 5' monophosphate: a possible indicator of premalignant changes in the large bowel. Cyclic adenosine 3', 5' monophosphate (cyclic-AMP) has been estimated in mucosal biopsy samples removed from the descending colon and rectum at endoscopy to investigate the possibility of using this substance for monitoring pre-malignant changes in the large bowel. Four groups of patients have been studied: those with normal large bowel and rectal mucosa; those with non-malignant inflammatory bowel disease; those with an adenomatous polyp in the descending colon or sigmoid colon; and those with a rectal adenocarcinoma. No difference was found in the cyclic-AMP content of 'normal' rectal mucosa, 'normal' colonic mucosa, 'diseased' colonic mucosa, carcinomas, and uninvolved mucosa adjacent to the polyps. Less cyclic-AMP was found in the polyps than in adjacent uninvolved mucosa. Conversely, more cyclic-AMP was found in the carcinomas than in adjacent uninvolved mucosa. It is concluded that although cyclic-AMP may be a very useful parameter for delineating the extent of the disease in individual patients, it is not a suitable biochemical marker for the screening of neoplastic changes in the large bowel in the population as a whole.


Specific methods for the determination of radioactivity in D-(-)-3-hydroxybutyrate in blood plasma. Two simple, high-yield rapid methods with good reproducibility are described, which permit the determination of radioactivity in plasma D-(-)-3-hydroxybutyrate. The compound is converted to acetoacetate, using a modified enzymatic method. In procedure 1, acetoacetate is reacted with 2,4-dinitrophenylhydrazine; the resulting hydrazone is oxidised by means of a sample oxidiser, and the product 14CO2 is collected in scintillation liquid and counted. In procedure 2, a Conway microdiffusion unit is applied. The acetoacetate is decarboxylated to acetone in the presence of o-phenylenediamine, and the acetone is then diffused into semicarbazide solution. This solution, containing the semicarbazone derivative of labelled acetone, is transferred to liquid scintillation and counted. In both procedures the radioactivity is measured simultaneously in a separate sample which was not subjected to the enzymatic conversion of D-(-)-3-hydroxybutyrate. The difference in radioactivity between the two samples is attributed to labelled (D-(-)-3-hydroxybutyrate.


Enzyme immunoassay: a review.

Enzyme immunoassay: a review. In the last few years, the use of enzyme labels in immunoassays has been investigated. The aim of this review is to evaluate critically the role of such labels in clinical biochemistry. Special attention has been given to the problems involved in preparing enzyme labels and the ways in which such labels can be used in a variety of heterogeneous and homogeneous assay systems.


Bioaccumulation and depuration of enteroviruses by the soft-shelled clam, Mya arenaria. Low levels of feces-associated natural virus, simulating virus numbers estimated to exist in moderately polluted shellfish-growing waters, were used to evaluate the effectiveness of depuration as a virus depletion procedure in soft-shell clams. Depuration effectiveness depended upon the numbers of virus bioaccumulated and whether virus was solids associated. Virus uptake was greatest when viruses were solids associated and pollution levels were equivalent or greater than those likely to be found in grossly polluted growing waters. Virtually all bioaccumulated feces-associated natural virus was deposited within either the hepatopancreas or siphon tissues. Viruses usually were eliminated within a 24- to 48-h depuration period. Dependence upon depuration of clams to elimate health hazards of virus etiology involved a risk factor not measureable in the study. The greatest reduction of health risks would come from the routine depuration of clams harvested from growing waters of good sanitary quality.


Classification of tumors of testis.

Classification of tumors of testis. In adult patients, testicular tumors consist predominantly of seminoma, embryonal carcinoma, teratoma, choriocarcinoma and mixtures of these. In infants and children, yolk sac tumor and teratoma are the usual tumors; in older age patients, it is predominantly spermatocytic seminoma and malignant lymphoma, although the others may occur as well. Leydig and Sertoli-granulosa cell tumors occur in all ages. The introduction of tumor markers and the capability to demonstrate these in tissue are the most important recent developments. However, there is great need for correlation of histology with these markers and histological demonstration of them.


Survey of human enterovirus occurrence in fresh and marine surface waters on Long Island. A variety of surface water systems, including a lake, a creek, and two marine embayments, were analyzed on a monthly basis for indigenous human enteroviruses and coliform bacteria. Findings are discussed in terms of the probable pollution sources to each system and their relationship to data from previous studies.


Assessment of cellular immune response to cancer of the breast. Cellular immune competence and cell-mediated immunity to tumor antigens have been studied in patients with breast cancer. Some patients have been shown to have depressed lymphoproliferative responses to phytohemagglutinin and in mixed leukocyte culture. In some cases, this depression appeared attributable to suppressor cells. Many patients with breast cancer had a cellular immunity to extracts of autologous or allogenetic tumors, as detected by lymphoproliferation and leukocyte migration inhibition assays. In addition, some breast cancer patients reacted to antigens associated with murine mammary tumor virus. Some of the tests for cellular immunity have revealed correlations with clinical course and, therefore, may be of use in the management of patients with breast cancer.


Mode of initiation of cell infection with sludge-associated poliovirus. Association with sludge solids did not allow poliovirus to enter cells other than through the normal receptor-mediated route. This implies that the infectivities of sludge-encapsulated virions are masked until their exposure permits binding to cell surface receptors.


Familial hypobetalipoproteinaemia in 9 children diagnosed as the result of cord blood screening for hypolipoproteinaemia in 10 000 Danish newborns. Out of 10 440 children, 266 with low (less than 2.5 centile) values for very low-low density lipoproteins in cord serum were chosen to be followed up to find out how many came from families with familial hypobetalipoproteinaemia (FHBL). In 176 families (66% of 266 families), FHBL was diagnosed in 9 children and their families.


Fate of herpes simplex virus in lymphocytes from inflammatory joint effusions. i. Failure of the virus to grow in cultured lymphocytes. The ability of lymphocytes isolated from the blood and synovial effusions of patients with rheumatoid arthritis to support the growth of herpex simplex virus (HSV) was studied. Whereas blood lymphocytes supported virus growth, effusion lymphocytes were usually non-permissive. Lymphocytes obtained from the effusions of patients with other forms of inflammatory arthritis or suspected viral arthritis commonly failed to support the growth of HSV, but the virus grew in effusion lymphocytes from patients with noninflammatory joint disease. Several nonspecific factors which might have accounted for this non-permissiveness were excluded.


Early changes in the chemical composition of the rat lung after silica administration. A group of 200 g male Sprague-Dawley rats was injected with 75 mg of quartz (less than 5 mu particle size) and changes in lung DNA, noncollagenous protein, total lipids, and collagen were studied after 6, 24, 72, 96, and 144 hr. Another group of rats received 10, 30, 59, and 75 mg quartz and the above lung analysis was performed 6 days later. Control rats received saline only. Both sets of experiments indicate that striking changes in the above parameters occur very early. The sequence of statistically significant changes was: lung weight (24 hr), DNA (24 hr), noncollagenous proteins (72 hr), total lipids (72 hr), collagen (144 hr). At the dose of 30 mg quartz/lung all the above parameters were significantly increased within 6 days after the lung injury. It is proposed that in early stages of experimental silicosis an excessive amount of collagen accumulates in the lung. Later, some of the deposited collagen is resorbed. This indicates that in the course of the silicotic fibroproliferative inflammation, the balance between collagen deposition and degradation varies.