Salt extraction of chromatin from normal diploid and SV-40 transformed human fibroblasts. Regulation of cell proliferation is a complex but exceedingly interesting form of gene expression. Overwhelming evidence in the past ten years has shown that the flow of cells through the cell cycle, and the regulation of cell population  density, are under the control of the eukaryotic genome. Changes in the function and structure of chromatin have been reported when G0 cells are stimulated to proliferate and when continuously dividing cells move through the different phases of the cell cycle from G1 to S, G2 and mitosis. The changes occurring in the structure and function of chromatin seem to be related to changes occurring in the chromosomal proteins, especially non-histone chromosomal proteins and chromatin-bound protein kinases. Studies are now in progress to isolate the genes, with their respective proteins, that regulate the different steps of the cell cycle.


Immunological disorders and malignancies in five young brothers. Five brothers of from 6 to 18 years of age experienced immunological or neoplastic disorders during an 8-year interval. 2 boys succumbed to glioblastoma multiforme, another to metastatic carcinoma, and the 2 surviving brothers had a histiocytic lymphoma and idiopathic thrombocytopenia purpura, respectively. The mother of the boys was healthy, but her twin sister died in utero of birth defects. We suggest that an intrinsic cellular defect inherited from their mother rendered the boys vulnerable to oncogenesis.


[On the biologic phenomenon of epidermotropism (author's transl)]. The biologic phenomenon of epidermotropism is mirrored in the dynamic equilibrium between different cell types within the epidermis. In various pathological conditions, this balance may be upset, or foreign epithelial, neuroectodermal and mesenchymal tissue may selectively infiltrate epidermal or other epithelial structures. The importance of the concept of epidermal symbiosis in these conditions and resulting similarities in the clinical picture and biologic behavior are pointed out.


The impact of long term estrogen support after hysterectomy. A report of 1016 cases. 1016 Women were placed on estrogen support following hysterectomy and have been followed for a total of 14,318 patient/years. Support was principally conjugated estrogen with the customary dose 1.5 mg/day. The follow-up studies in this group of women shows a marked drop in deaths from all causes over those which might have been expected. This improvement in mortality is principally the result of diminished number of deaths from heart attack and from cancer. Those causes of death which would not be expected to be related to hormonal therapy show the anticipated number of mortalities. There is also a marked improvement in the clinical evidence of osteoporosis. In this group there is an increase in the number of breast cancers over those which might have been expected, but there is a lower mortality from breast cancer than the anticipated mortality. The general impact of long-term estrogen therapy following hysterectomy is favorable over those figures for the expected incidence of cancer and heart disease.


Clinical significance of circulating C-peptide in diabetes mellitus and hypoglycemic disorders. Proinsulin is converted to insulin and C-peptide in the pancreatic in the pancreatic beta cells: the latter two peptides are secreted in equimolar concentrations. Thus, measurements of serum C-peptide provide a means of assessing pancreatic beta cell function in addition to that of insulin. This technique has proved particularly useful in insulin treated diabetic patients in whom the development of circulating insulin antibodies interferes with the radioimmunoassay of the hormone. The C-peptide assay has also been used to facilitate the diagnosis of various hypoglycemic conditions, including islet cell tumors and factitious injection of insulin. The extraction of C-peptide in the urine reflects average serum values over a period of time and urine C-peptide measurements are especially useful in children or individuals in whom repeated blood sampling is difficult.


Single units activities in ventral posterior and posterior group thalamic nuclei during nociceptive and non nociceptive stimulations in the cat. The purpose of this study was to define, in hyperventilated and unanesthetized cats, the role of the posterior thalamic nuclei in pain mechanisms. Unit activities of these structures were compared to those of the ventro-posterior nucleus during non-noxious (touch, brushing) and noxious stimulations (pinches and intra-arterial injections of bradykinin into the limbs). 135 cells with somatic inputs and clear peripheral excitatory receptive field were studied. The cells driven by noxious stimulations were located in the posterior group nuclei as anatomically defined by Rinvik. These units, preferentially excited from contralateral receptive fields, were localized in POm, POl, suprageniculate nuclei, the magnocellular division of the medial geniculate body (Mgmc) and the ventral part of the lateral posterior nucleus. At this level two groups of units were found: those driven only by noxious stimulations and those driven by both noxious and non-noxious stimulations. On contrast, cells recorded at the levels of the VPm and VPl were not activated by noxious stimuli. These results emphasize the role of the posterior thalamic nuclei in pain processing.


Adenoid cystic carcinoma of the breast. Light and electron microscopy and a brief review of the literature. The case of two patients with adenoid cystic carcinoma of the breast are reported. In case 1, the predominant pattern was that of a solid, basaloid, cylindromatous tumor, whereas case 2 showed the typical adenoid cystic appearance. Results of electron microscopical study confirmed that many of the cystic spaces seen by light microscopy were extracellular compartments enclosed by tumor cells. Many of the tumor cells contained densely packed fibrils, which support the myoepithelial origin of these tumors. Approximately 92 well-documented reports of this lesion are briefly reviewed.


Pseudoadenoid cystic carcinoma of the breast. An example of a cribriform intraductal carcinoma that closely resembled adenoid cystic carcinoma is described. The true nature of the tumor was revealed by electron microscopy and the case is used to demonstrate that the diagnosis of this tumor cannot be safely made by routine histologic techniques alone. Review of conflicting views othe value of mucin histochemistry expressed in the literature suggest that ultrastructural examination is a useful alternative tool in the differentiation of adenoid cystic carcinoma of the breast from cribriform intraductal carcinoma.


Tubular carcinoma of the breast. Clinical, histological, and ultrastructural observations. Tubular carcinoma is a neoplasm with good prognosis of features that warrant its differentiation from other mammary carcinomas and from sclerosing adenosis. Cases of nine primary unilateral neoplasms are presented and ultrastructural observations in three discussed. The mean diameter was 1.2 cm and the mean patient age was 48.7 years. None of the patients died and only one of six had residual tumor in the mastectomy specimens. Three patients had an excisional biopsy, one followed by radiation. All biopsy specimens showed infiltrating angular tubules with single-lining epithelium, minimal anaplasia, rare mitosis, and scant or absent myoepithelium. The tumor cells related to each other by desmosomes, cytoplasmic interlocking processes, and tight junctions. Myoepithelial cells were rare. Since tubular carcinomas are distinct variety of ductal epithelial neoplasms of the breast, and are well differentiated with low malignant biological behavior, a less radical approach may be the treatment of choice.


Paget disease of the male breast.

Paget disease of the male breast. Twenty-two fully documented cases of Paget disease of the male breast are reviewed, and an additional patient is reported. The most common initial symptom was ulceration and excoriation, while a breast mass was palpable in the majority of patients. Nipple changes were surprisingly rare, in contrast to the female. The prognosis of the disease in the male appears to be worse than in the female, with the Paget carcinoma carrying a worse prognosis than the "ordinary" male breast cancer. The subareolar location and the meager volume of tissue interposed between the tumor and chest wall may be an important factor in this regard. Nipple changes or symptoms (ulceration, discharge, enlargement) are mroe apt to be due to cancer in the male than in the female. Therefore, prompt diagnosis is mandatory.


Progesterone receptor in cystosarcoma phyllodes. A specific receptor for progesterone has been found in a cystosarcoma phyllodes, as determined by charcoal adsorption and sucrose gradient analysis. Similar assays for estrogen receptors were negative. The tumor consisted almost entirely of stroma that contained the progesterone receptors. The epidemiology and natural history of cystosarcoma do not strongly support the hypothesis that it is controlled by female sex hormones, but the presence of the progesterone receptors suggests that some cystosarcomas are hormonally regulated, and thus may be responsive to therapeutic hormonal manipulation.


Surgical management of hepatic vein occlusion by tumor: Budd-Chiari syndrome. We report new operative approaches to the treatment of hepatic vein occlusion due to malignant tumors in the liver and their results in four patients. Two patients had hepatoma, one had metastatic melanoma, and one had metastatic leiomyosarcoma. All of them had abdominal pain, abdominal distention secondary to ascites, and massive hepatomegaly. The right lobe and medial segment of the left lobe of the liver were involved in three patients, and the involvement was diffuse throughout the liver in one. Hepatic veins were occluded completely in one patient, and two of three veins were occluded in the others. Two patients were treated by hepatic resection and removal of tumor thrombus from the hepatic vein under isolation-perfusion technique. They lived 18 and six months, respectively, without recurrence of Budd-Chiari syndrome. Tumors in the other patients were diffuse and could not be resected. The hepatic artery was ligated and chemotherapy was given postoperatively. Ascites and abdominal pain disappeared completely in one, who survived 17 months. The other patient had significant palliation and lived nine months.


IgM antibodies to cytomegalovirus during pregnancy. IgM antibodies to cytomegalovirus (CMV) could be detected in about 7 per cent of 629 pregnant women whereas in 225 nonpregnant control women of similar age distribution only 2.6 per cent showed CMV IgM antibodies. Intrauterine CMV infections were almost exclusively detected among the CMV IgM positive gravides. The high incidence of CMV IgM antibodies in pregnant women can be possibly explained by an increased rate of CMV reactivations during pregnancy. We were able to show that during CMV reactivation an intrauterine infection might occur.


Herpesvirus group antibodies in children with Hodgkin's disease. During the period of three years ((1972-1974), serum samples from 60 patients (children and adolescents) with lympho-hematopoietic system diseases were examined for antibodies to all four human herpesviruses. Among these were 26 active Hodgkin's disease (AHD) patients and 6 HD patients with a minimum five years' remission. Simultaneously matched controls (age, sex) of AHD patients were examined. Antibody levels against the viral capsid antigen of Epstein-Barr virus (EBV/VCA) in AHD patients were significantly higher, with overrepresentation of higher titres (greater than or equal to 1:160), than in matched controls. The lowest EBV/VCA antibody titres were in the leukemia-non-Hodgkin's lymphoma patients. We could not prove any significant relationship between cytomegalovirus or herpes simplex virus type 1 antibody titres and AHD or any other disease of lympho-hematopoietic system. The varicella-zoster virus antibody titres in AHD patients were significantly higher than in matched controls. No significant differences in antibodies against EBV/VCA and the other human herpes viruses between the evolution and remission period of AHD patients could be detected. No differences in EBV/VCA antibody titres were observed between the healthy school-children aged 10 to 15 years who were and who were not in contact with a HD patient.


A comparative electrophoretic examination of swine vesicular disease virus and Coxsackie B5 virus. Purified suspensions of Coxsackie B5 virus and swine vesicular disease virus (SVDV) were prepared by harvesting and purifying cell pack virus. Crossed immunoelectrophoresis was carried out with purified N and H antigen fractions (full and empty particles). Relative migration velocity (RMV) was calculated for the N antigen fraction of 3 SVD viruses (UKG72, HK71 and Italy 66) and 2 Coxsackie B5 viruses (Faulkner and 8068). The prototype strain of Coxsackie B5 virus (Faulkner) had a low RMV almost identical to that of the first isolated strain of SVDV, Italy 66. A recent isolate of Coxsackie B5 virus (8068, isolated in UK in 1973) had a relatively high RMV very close to that of the UKG72 strain of SVDV isolated in the UK in 1972. Also the Hong Kong strain of SVDV (HK71) had a high RMV value. These observations are considered in relation to the emergence of swine vesicular disease.


[Histochemical study of the phosphomonoesterases of cells producing steroid hormones]. The results of detection of phosphatases with different substrates in the cells forming steroid hormones in ovaries, testes and adrenals of newborn seals, ovaries of adult seals and adrenals of adult rabbits, dogs and man were compared. The first group of substrates includes sodium glycerophosphate, glucose-1-phosphate, nitrophenylphosphate (Gomori's sulfide method). The second group includes naphthol phosphates AS-BI, AS-MX and AS-BS, whose hydrolysis is determined by simultaneous and successive azocoupling with diazotized benzidine and stable diazotates by fast blue and fast cherry-coloured. Detection of phosphomonesterases in the cells forming steroid hormones with these two groups of substrates gives absolutely different results which are presented in the table. These results are thought to be associated with presence of the isoenzyme, intensively splitting azotholphosphates, but inhibited by the diazonium salt. This isoenzyme is found mainly in the cells producing steroid hormones.


[Morphogenesis and features of the pathogenesis of adenovirus and chlamydial infections of the conjunctiva]. Morphogenesis of adenovirus (epidemic keratoconjunctivitis) and chlamydia (paratrachoma) infections was studied on the material consisting of scrapings of the conjunctiva and biopsy specimens taken from 410 patients, by histological, cytological, histo- and cytochemical electron-microscopic and immunohistochemical methods. The investigations conducted showed that the morphogenesis of adenovirus and chlamydia infection of the conjunctiva was underlaid by a complex of pathological disorders including changes in the epithelium, where the cytopathogenic effect of the pathogene was realized, and changes in the conjunctiva tissue proper, the latter being transformed in these infections into the organ of immunogenesis.


[Several features of cardiac lipid metabolism under conditions of diphteritic intoxication]. Basing on the collation of data of histochemical and electron-microscopic investigations of the myocardium of rabbits under conditions of diphtherial intoxication and in blocade of the lipolysis processes in the intact and damaged heart, the authors singled out three ways of utilization of lipids by the cardiac muscle. The first way is associated with adsorption of fatty acids from the circulating blood. This way depends on catecholamines and is blocked with nicotinic acid. The second way is determined by splitting of myocardial triacylglycerols. It does not depend on concentrations of catecholamines and can be blocked only following prolonged exposure to nicotinic acid. The third way is associated with intensity of functioning of mitochondria; destruction of the latter brings about liberation of membrane phospholipids which later are transformed into nonesterified fatty acids. This way does not depend on catecholamines and can be inhibited by acid lipids accumulating in the myocardium as a result of impairment of utilization of these lipids.


[Current trends in the study of phagocytosis and non-specific resistance]. Phagocytosis begins with exocytosis--"extrarapid" discharge of bactericidal proteins and factors of permeability into the extracellular medium. A viewpoint was put forward on an "avalanch-like" character of the outcome of cationic proteins from leukocyte granules in inflammation and their participation in formation of a nonphagocytic type of resistance. In phagocytosis bacteria perish due to the myeloperoxidase system, lysozyme,  lactoferin and nonenzymic cationic proteins. Hereditary deficit of the above-mentioned substances leads to intraleukocytic microbicidal insufficiency, a drastic decrease in the nonspecific resistance of the organism and to development of fatal granulomatous disease, and to other forms of pathology associated with genetic defects of the bactericidal systems of leukocytes.


[Russel bodies in the duodenal mucosa in ulcer disease]. A total of 103 specimens of the duodenum mucosa incised during the resection of the stomach in different variants of the course of peptic ulcer were studied. It was shown that Russell's bodies (RB) in the mucosa's layer proper were formed by cellular elements analogous to immunoglobulinocytes of the lymph nodes and spleen of the laboratory animals which had been subjected to an experimental stimulation of immunogenesis. Peculiar patterns of contacts of RB with the mast cells and possible significance of these contacts are discussed. On the basis of the data obtained the conclusion was drawn that elimination of the RB substrate in the duodenum mucosa was realized in two directions: a) centripetal by penetration of the substrate into the lymphatic and blood vessels; and b) centrifugal, by its evacuation into the lumen of the alimentary canal.


[Genetic syndromes of multiple congenital developmental defects]. Analysis of developmental defects in 615 children who had died in hospitals in Minsk during the 5-year period (1971--1975)showed that in 274 children defects were multiple. This number includes 78 children with chromosomal diseases and 37 children with genic syndromes of multiple congenital malformations (MCM). The total number of genic syndromes of MCM is extremely great, at present over 250 of them are known. Analysis of the available literature and the authors' own findings made it possible to develop criteria of morphological diagnosis of genic syndromes of MCM with due account for different diagnostic significance of various defects. Genealogical studies and estimation of population incidence of individual malformations are also necessary for diagnosis of genic syndromes of MCM and identification of new syndromes. An etiological classification of MCM and that of genic syndromes of MCM are considered on the basis of isolation of similar clinical forms, which are of important for differential diagnosis.


[Tumors of the liver, biliary tracts and pancreas in monkeys]. The article deals with spontaneous tumours of the liver, biliary tract, and pancreas observed in monkeys of the Sukhumi Simian Nursery during the period of 1960--1974. Among tumours of the liver hepatoadenomas predominated: they were revealed in 4 macaco rhesuses, one green marmoset. In a young baboon hamadryad adenomatous hyperplasia was observed to be developed against the background of hepatocirrhosis. All three cases of tumours of the biliary tract in baboons hamadryad were classified as adenocarcinomas. Descriptions of liver cell anaplastic carcinoma with extensive metastasizing in Macaco rhesus, those of insuloma of the pancreas in the same species, and of papilary duct carcinoma of the head of the pancreas in a red ape (Erythrocebus patas) are presented for the first time. All the monkeys with tumours, but one, were at the age over 10 years and were born in the Nursery, or had been brought to it more than 7 years before.


Blindness of cerebral origin in acute intermittent porphyria. Report of a case and postmortem examination. Sudden permanent blindness of cerebral origin, in addition to severe abdominal pain, hypertension, convulsions, and peripheral neuropathy developed in a 21-year-old woman, a victim of acute intermittent porphyria. Findings of the pathological examination of the brain showed extensive infarction in both occipital lobes. The pathological changes were consistent with anoxia. We discuss and review the literature of the possibility of "vasospasm" of both posterior cerebral arteries. Follow-up studies with serial EEG showed either focal epileptogenic activity or diffuse slow waves. The most consistent epileptic discharges were found in the occipital regions. The favorable response to the treatment of seizures with carbamazepine in this patient might encourage further clinical trials.


Effects of a single methylprednisolone dose on a facilitatory response of mammalian motor nerve. Long-term glucocorticoid dosing directly enhances a facilitatory function of cat soleus motor nerve terminals. Posttetanic potentiation (PTP) of soleus contraction is a manifestation of this prejunctional facilitation. The present study demonstrates that the same enhancement of facilitation is produced with a single large intravenous dose methylprednisolone. The single dosing method, however, showed an initial suppression of facilitation that neared recovery in four hours. Thereafter, the characteristic augmentation of prejunctional facilitation emerged, peaking in 24 hours. Return to control required four days. Knowledge of this time course enabled centrally disconnected motor nerve endings to be identified as the site of both phases of the steroid action. Since the neuromuscular facilitation studied is equivalent to that triggered by neostigmine-like drugs, the results infer that the antimyasthenic effect of glucocorticoids may involve a direct action on motor nerve endings.


Antimyasthenic action of corticosteroids. Tests were made in vitro fo the action of prednisolone on nerve backfiring, muscle twitches, tetanus decay rates, miniature end-plate potential amplitude and frequency, and the block induced by curare, hemicholinium, and excess magnesium. At about 0.1 mM concentration, the steroid showed no 'veratrinic' or decurarizing action, and the probability of transmitter release was not increased in 10mM Mg++. Moreover, when acetylcholine stores were depleted in hemicholinium, there was no evidence of repair by steroid. The results are taken to mean that prednisolone and congeners do not owe their therapeutic efficacy in myasthenia to actions at the neuromuscular junction. It is therefore concluded that the clinical benefits from steroids are related to systemic, possibly immunosuppressive effects.


Characteristic early electroencephalographic changes in herpes simplex encephalitis. We review electroencephalograms taken from 17 patients with severe meningoencephalitis within seven days of onset of CNS symptoms and prior to cortical brain biopsies. All patients had CNS disease clinically compatible with the diagnosis of herpes simplex encephalitis (HSE). The diagnosis was demonstrated by the isolation of virus from the brain in five patients (group 1) but considered highly unlikely in the other 12 patients (group 2) by negative immunofluorescent studies and failure of viral isolation from the brain tissue. Abnormal but nonspecific EEGs with diffuse or focal slowing were found in all patients. Distinctive high-voltage, 1-cycle-per-2-to-3 seconds periodic sharp waves from unilateral temporal lobes were seen only in three of the five patients with virologically proved HSE but in none of the 12 patients without viral isolation. This EEG pattern is strikingly similar in all three patients, regardless of their age, and may be specific for the early diagnosis of HSE prior to brain biopsy. The EEGs of the other two patients with proved HSE did not contain such abnormalities. Athough periodic EEGs with some resemblance to those previously described may occur in other CNS disorders, their presence strongly suggests the diagnosis of HSE when recorded from patients with viral meningoencephalitis. Moreover, EEGs may help locate the best site for cerebral biopsy since maximal yield of the virus in this study was from unilateral temperoal lobes corresponding with the site of local EEG changes.


Increased cellular responsiveness to epinephrine in primary open-angle glaucoma. The concentration of I-epinephrine hydrochloride necessary to inhibit lymphocyte transformation by 50% (I50 value) was determined in vitro for nine patients with primary open-angle glaucoma (POAG) and in seven controls. The lymphocytes of the patients with POAG were significantly (P less than .05) more responsive. This result is consistent with the hypothesis that patients with POAG have an increased responsiveness to agents that elevate intracellular cyclic adenosine monophosphate levels.


Watery diarrhoea (WDHA) syndrome associated with carcinoma of the lung. A patient with bronchial carcinoma presented with profuse watery diarrhoea, hypokalaemia and hypochlorhydria. Laparotomy appeared to exclude the usual cause, a pancreatic islet-cell tumour. Relief of the diarrhoea was achieved only with prednisone therapy. Evidence is adduced to implicate a humoral factor produced by the tumour, although in this case, the serum VIP level was not elevated.


Enzyme histochemistry of rat folic acid nephropathy. One hour after a single i.v. dose of 250 mg/kg folic acid, the straight portion of distal tubules in the outer medulla of rat kidneys showed a distinct reduction in succinate dehydrogenase, NADH2-diaphorase, glutamate dehydrogenase, cytochrome oxydase, Na+/K+-ATPase, and acid phosphatase activity. In contrast, the proximal tubules exhibited only a reduction in glutamate dehydrogenase and alkaline phosphatase activity. At this time the straight portion of the distal tubules, whose enzyme activity had changed, showed partly regressive epithelial changes. 24 hours after folic acid administration an even greater reduction in enzyme activity had occurred in the straight portion of distal tubules; these structures also became dilated. The adjacent collecting tubules and the corresponding proximal tubules were also severely dilated, the proximal tubules showing a loss in enzyme acitivities similar to those observed in the distal tubules. 48 hours after folic acid administration the changes largely resembled those observed after 24 hours, but were more pronounced. At this time a tubular regeneration was observed. 72 hours after folic administration extensive normalization of the histological and histochemical changes had occured. It is postulated that a disturbance of the hairpin counter-current mechanism occurs as a result of a direct, concentration-dependent effect of folic acid on the enzymes of the energy supplying metabolism. A dilation in the region of the loop of Henle and the collecting tubules occurs subsequently.


Evidence for head-head interactions in myosin from cardiac and skeletal muscles. 1. Binding of MG-ADP to both heart and fast skeletal myosin was found with 3 methods to proceed in 2 steps. One mole of MG-ADP binds with high affinity (K approximately equal to 10(6) M-1) and subsequently a second with lower affinity (K approximately equal to 10(2)-10(4) M-1) per myosin. Only one mole of MG-ADP was found to bind with the high affinity to isolated myosin heads. This implies that binding of MG-ADP to intact myosin exhibits negative cooperativity. 2. When a nucleotide is bound, the 2 heads of a single myosin molecule adopt different conformations since on each head a different type of essential thiol group was found to be the most reactive towards N-ethylmaleimide. In the presence of MG-pyrophosphate a thiol-1 is the most reactive essential group in both heads. Therefore, the nucleoside moiety seems to be required for this latter type of head-head interaction.


[Dose-dependent effects on phenobarbital on microsomal liver enzymes of chicken embryos (author's transl)]. Levels of cytochrome P-450 and the activities of amino-pyrinedemethylase and p-nitrophenol-UDP-glucuronyltransferase were measured in homogenates and microsomes of 16 to 19 day old chicken embryos exposed in ovo to phenobarbital. The activities of glucose-6-phosphatase were measured on the 19th day of incubation. After the highest dose of 3 X 8 mg phenobarbital, cytochrome P-450 increased 3-6fold, aminopyrinedemethylase activity 7fold and the activity of p-nitrophenol-UDP-glucuronyltransferase 3fold. Glucose-6-phosphatase activity was not increased but decreased. Corresponding to the given dose of phenobarbital (3X3, 3X4, 3X6, 3X8 mg) into the yolk sac an increase in enzyme activity levels mentioned above could as a rule be demonstrated at the level of p less than 0.0025. Calculated microsomal protein amounted to 43.0+/-6.5 mg/g liver.


Myogenic foam cells in explants of fatty dots and streaks from rabbit aorta. Morphological studies. Present studies indicate that in explants of early atherosclerotic lesions removed from aortae of young rabbits on a 1% hypercholesterolemic diet for four and seven weeks respectively, myogenic foam cells (MCF's) were capable of emigrating into the culture medium and maintained their ability to produce microfibrils, elastic tissue elements, and collagen fibrils. In explants of the smallest lesions (fatty dots and small streaks) the MFC's divided prior to, or while emigrating. At the interphase between the primary tissue and the culture medium they contained in intracytoplasmic vacuoles fragments of elastic tissue and extraneous substances which were reminiscent of cellular debris. It is possible that this phenomenon represents a true phagocytic property of the MFC's. All formed extracellular connective tissue components were also produced by the emigrated MFC's in the tissue surrounding the cellular outgrowth. In the large fatty streaks cell division was observed at the interphase between the tissue and culture medium, but not within the substance of the explant; here cellular necrosis was prominent. The fat inclusions in the MFC's of explants and the outgrowth had the appearance of conglomerateds of unorganized, and only at times concentric, membranous profiles rather than that of homogeneous droplets observed by electron microscopy in these cells in tissue sections. In the outgrowth from explants of normal aortic areas adjacent to the lesions a moderate number of smoot muscle cells contained fat inclusions; these were almost totally absent in cells of the primary aortic cultures from normal aortae. It is conceivable that the migratory and phagocytic properties of the MFC's observed in the present study relate to some aspects of regression of atherosclerotic lesions; this, however, remains highly speculative at present.


Cholesterol ester accumulation in cultured aortic smooth muscle cells. Induction of cholesterol ester retention by chloroquine and low density lipoprotein and its reversion by mixtures of high density apolipoprotein and sphingomyelin. Accretion of cholesterol ester was studied in rat aortic smooth muscle cells in culture. Confluent multilayers of smooth muscle cells were exposed to human low density lipoprotein (LDL) and chloroquine and this treatment resulted in a very marked increase in cellular cholesterol ester. The degree of enrichment in cholesterol ester was related inversely to the cell density in the petri dish and was maximal in 48 h. The morphological changes after 48 h incubation with chloroquine and LDL consisted of accumulation of numerous membrane-bound inclusions containing electron-dense and electron-lucent material, some of which resembled secondary lysosomes. These changes resembled some of the changes observed in human and experimental atheromatosis. Similar inclusions were seen also in cultured human skin fibroblasts which accumulated large amounts of cholesterol ester during 48 h incubation with LDL and chloroquine. Removal of the accumulated cellular cholesterol ester was studied in the two cell types and it was markedly enhanced in the presence of lipoprotein-deficient serum and high density apolipoprotein-sphingomyelin mixture. The morphological findings after 24 h of post incubation revealed the presence of empty vacuoles, membrane whorls and cytoplasmic lipid droplets. The present results indicate that aortic smooth muscle cells in culture can serve as a good model to study the role of the lysosomal system in atherogenesis.


Effect of lipoprotein-X on hepatic cholesterol synthesis. The effect of different lipoproteins (lipoprotein-X and lipoprotein-B; LP-X and LP-B) on hepatic cholesterol synthesis was studied in vivo in rats. Lipoproteins were continuously infused into rats for 16 hours so that 24 mg cholesterol/100 g body weight were applied. Serum cholesterol level was nearly doubled after the infusion period. Lipoprotein electrophoresis revealed the predominance of the infused lipoprotein in the serum. LP-B infusion caused a reduction of cholesterol synthesis (42% of control values) and reduced the increased cholesterol synthesis of bile fistula rats to values below normal. LP-X did not reduce hepatic cholesterol synthesis significantly nor did it normalize the enhanced synthesis following biliary diversion. However, hepatic free cholesterol concentration increased after LP-X infusion. The effect of LP-X on liver cholesterol synthesis is similar to that of lecithin: cholesterol dispersions. The failure of LP-X to exert a feedback inhibition on cholesterol synthesis may therefore contribute to the mechanism of hypercholesterolemia in obstructive jaundice.


Pharmacokinetics of clofibrate in familial hypercholesterolemia. Some patients with familial hypercholesterolemia (FHC, type II) are highly responsive to the cholesterol-lowering effect of clofibrate, while others are not only resistant to this effect but may even show an increase in plasma beta-lipoproteins. In an attempt to find an explanation for these striking differences, we have studied the pharmacokinetics of clofibrate in FHC patients at both extremes of responsiveness. The results disclosed several major differences between the two groups. Plasma clofibric acid (CPIB) measured during the chronic administration of the drug was significantly higher in the responders than in the non-responders, whether all patients in each group or only those with tendon xanthomas were considered. Plasma CPIB concentrations were negatively correlated with body weight in the responders but not in CPIB-resistant patients. They were also inversely proportional to decreases in plasma beta-lipoprotein cholesterol after chronic clofibrate administration in the responsive group, but directly proportional to increases in the non-responders. Increasing the dose of clofibrate from 2 to 3 g/day in CPIB-resistant patients always resulted in an increase in plasma CPIB levels, but this was followed in some patients by a decrease and in others by an increase in plasma beta-lipoprotein cholesterol concentrations, so that the overall effect was not statistically significant. The half-life of plasma CPIB was measured over 48 h after a single 1-g dose of clofibrate in patients who had not received this drug for at least 3 weeks. Half-life was significantly longer in the responsive patients. In addition, the bioavailability and the rate of absorption of clofibrate tended to be higher in this group than in the resistant patients. We suspect that both groups differ not only in the metabolic handling of clofibrate but also in some aspect of their beta-lipoprotein cholesterol metabolism.


Treatment of hypertriglyceridemia with metformin. Effectiveness and analỳsis of results. The triglyceride-lowering effect of metformin (N,N-dimethylbiguanide) was tested in a series of patients with stable hypertriglyceridemia (types IIB, III and IV) and with variable degrees of glucose intolerance. Metformin caused a 38% mean decrease of plasma triglycerides. A selective decrease of very low density lipoprotein cholesterol was observed without reciprocal increase of low density lipoproteins. Thirty patients completed the study. Eighteen, who showed a hypotriglyceridemic effect exceeding 30%, were considered as "Responders"; the other 12, where the effect was negligible, were considered as "Non-Responder". Analysis of the pre-and post-treatment glucose tolerance tests of Responders and Non-Responders showed that the former had, on the average, a normal glucose tolerance and insulin secretion, whereas the latter had an impaired glucose tolerance with increased insulin secretion. These parameters were only slightly modified by metformin. The conclusions of this study support the hypothesis that biguanides exert a triglyceride-lowering effect by decreasing lipoprotein secretion, independent of changes in glucose tolerance and/or insulin secretion.


On the rise in low density and high density lipoproteins in response to the treatment of hypertriglyceridaemias in type IV and type V hyperlipoproteinaemias. In Type V hyperlipoproteinaemia the concentration of LDL and HDL cholesterol is low. When the hypertriglyceridaemia is normalized, either by diet or micotinic acid, both LDL and HDL increase. In Type IV hyperlipoproteinaemia both LDL and HDL cholesterol decrease with increasing VLDL levels. During treatment of Type IV the change in LDL cholesterol is linearly related to the pretreatment LDL concentration so that higher LDL levels will fall and the lower will rise. HDL levels will also rise. The fall in VLDL during treatment is rapid and the rise in LDL also occurs rapidly indicating a relationship between these two reciprocal changes. HDL cholesterol, however, however, remains constant some time after VLDL has reached its lowest level and the rise occurs later indicating another mechanism than for LDL. These changes in the three major lipoprotein classes deserve clinical attention. While both the fall in VLDL and the rise in HDL may be benefical from the point of view of atherosclerosis the rise in LDL may be harmful. These is at present no way to evaluate the effect of these complex changes. However, these findings stress the imporance of considering changes in lipoprotein levels and not only in total serum triglycerides and cholesterol during treatment of hyperlipoproteinaemia.


Effect of estrogens on the concentration and composition of arterial sterols and steryl esters in male white carneau pigeons. The effect of short-term (6 months) administration of conjugated equine estrogen (Premarin) on content and composition of the aortic sterols in male shite Carneau pigeons while they were on a cholesterol-free grain diet was investigated. Estrogen treatment resulted in a 38% increase (P less than 0.05) in free sterol concentration, with a 28.8% concomitant decrease (P less than 0.05) in the percent of cholesteryl esters. The total sterol concentration remained unchanged. This finding suggests that estrogens might influence the synthetic or hydrolytic (or both) processes that control the concentration of cholesteryl  esters in the aorta. Fatty acid composition of steryl esters did not change significantly. The cholesterol content of plasma showed a mild reduction (14%) whereas the triglycerides increased significantly (30%).


Hypertension-accelerated atherogenesis in cholesterol-fed rabbits. Entry of 125I-labelled low density lipoprotein ([125I]LDL) into the aortic intima was studied over 6 hours in normotenisve and hypertensive rabbits fed a 1% cholesterol diet for 9 and 4 weeks respectively. Studies were also made in hypertensive and normotensive cholesterol-fed rabbits in which blood pressure was reduced acutely with parenteral hydralazine. In all groups the entry of E1125I]LDL was greatest in the aortic arch and significantly less in both the descending thoracic and abdominal regions. Lipoprotein entry into the aorta of cholesterol-fed rabbits was increased some 10-fold over the corresponding value previously found in rabbits fed a normal diet [1]. This increase was due to increased vascular permeability as well as to increased plasma LDL concentration. The hypertensive cholesterol-fed rabbits did not show significantly greater entry of [125I]LDL than the normotensive cholesterol-fed rabbits. Comparison of the rate of LDL entry over 6 house and the quanitity of cholesterol accumulated in the aortic segments over the period of cholesterol feeding indicated that lipoprotein fractions other than LDL must contribute singificant amounts of cholesterol to the developing lesion. The finding that LDL entry paralledled accumulation during cholesterol feeding, together with the finding that acute reversal of hypertension did not reduce the entry of [125I] LDL suggest that mechanisms other than increased filtration of plasma low density lipoprotein contribute significantly to the accelerated development of atherosclerosis in hypertension.


Transplacental acquisition of cytomegalovirus antibody. Observations with two different CMV strains. Titrations of complement fixing antibody (c.f.a.) were done in paired sera of 127 women at delivery and cord blood of their babies using two different cytomegalovirus strains both of which had undergone many passages in WI-38 cells, With High strain 74% of the mother and 41% of the babies were negative at the serum dilution 1:4. With Towne strain the incidence of negative was 5% both among the mother and the newborns. Worth of mention is the study of correlation of the titers in the matched pairs of sera according to the strain. With High strain 23 of the babies had c.f.a. titers significantly higher than their mothers and 14 of them were from negative women. This situation could be suggestive of an active placental transfer of IgG globulins. No such disparities were found using Towne strain as antigen. 48 pairs of sera were then tested with both strains by indirect hemagglutination, indirect immunofluorescence and gel precipitin test. Results are discussed in relation to the possible differences in the antigenic composition of the two cytomegalovirus strains.


Complement fixing antibody to varicella-zoster virus in different age groups and after zoster. The titer of complement fixing antibody to Varicella-Zoster virus was studied in sera from 215 normal persons living in Padua and its province and from 39 Zoster patients. About 74% of sera from normal persons aged 21-80 years had antibody titers larger than or equal to 1/4. Lower titers were present in the age groups 1-20 years and 81-90 years. Sera from Zoster patients had high titers from the second week following the onset of the rash. They were still comparatively high until the fifth month, sometimes remaining detectable even after a year or more.


Neurobiology of aggressive behavior.

Neurobiology of aggressive behavior. Causality, neurological mechanisms, and behavioral manifestations may be heterogeneous in different forms of aggressive behavior, but some elements are shared by all forms of violence, including the necessity of sensory inputs, the coding and decoding of information according to acquired frames of reference, and the activation of pre-established patterns of response. Understanding and prevention of violence requires a simultaneous study of its social, cultural, and economic aspects, at parity with an investigation of its neurological mechanisms. Part of the latter information may be obtained through animal experimentation, preferably in non-human primates. Feline predatory behavior has no equivalent in man, and therefore its hypothalamic representation probably does not exist in the human brain. Codes of information, frames of reference for sensory perception, axis to evaluate threats, and formulas for aggressive performance are not established genetically but must be learned individually. We are born with the capacity to learn aggressive behavior, but not with established patterns of violence. Mechanisms for fighting which are acquired by individual experience may be triggered in a similar way by sensory cues, volition, and by electrical stimulation of specific cerebral areas. In monkeys, aggressive responses may be modified by changing the hierarchical position of the stimulated animal, indicating the physiological quality of the neurological mechanisms electrically activated.


[Peripheral nerve disease associated with acute renal failure due to bromate poisoning]. A case of 21 year old male with neuropathy caused by renal insufficiency was present. He had taken bromate (mixed powder of potassium bromate and sodium bromate) for the purpose of suicide and suffered from acute renal insufficiency and hard of hearing. Renal dysfunction improved gradually by peritoneal dialysis and hemodialysis. However, on the 32th day after the onset, burning pain appeared in the bilateral feets. Following this, he began to complain of the disturbances of superficial and deep sensory below the ankle jerks and the weakness of his toes. Considering the clinical features, we supposed that the disturbance of the peripheral nerve was caused by uremia due to taking bromate. N. suralis was biopsied on the 80th day after the onset and examined electron microscopically. Electroscopical findings was as follows. Degeneration of the Schwann cells and irregularity or destruction of the myelin sheaths were observed. The axoplasm of the myelinated nerve fiber were relatively preserved as compared with the changes of the myelin sheaths. In the unmyelinated nerve fibers, cavity formations were observed. The findings of regeneration were not observed. From the electron microscopical findings, we speculate that the changes of the Schwann cells and the myelin sheaths are primary resulting from the disturbance of the metabolism of the Schwann cells. We speculate that anemia and hypoproteinemia caused by bromate disturbed regeneration.


Immunological mechanisms in metastatic spread and the antimetastatic effects of C. parvum. The effects of the host's immune response on metastatic spread was investigated by observing the numbers of pulmonary metastases that developed from an s.c. implant of the Lewis lung carcinoma in C57BL mice in which different cell populations had been suppressed. Macrophage function was impaired by treatment with silica (Si), cortisone acetate (CA), or trypan blue (TB). T-cell function was depressed by adult thymectomy and sublethal irradiation, or by treatment with antilymphocyte serum (ALS). Metastasis was significantly increased and phagocytic activity decreased by Si and CA, but were unaffected by TB. Thymectomy and irradiation had no effect on metastases, whereas ALS when given before, but not after tumour growth, reduced their number. The antimetastatic action of the immunopotentiating agent C. parvum was investigated in these immunologically impaired mice. It was unaffected by Si, CA or TB. However, the inhibiting effect of these agents on phagocytic activity was overcome by treatment with C. parvum. Its antimetastatic action was unaffected in mice which had been thymectomized and irradiated, but could be abrogated by ALS. However, ALS was only able to prevent this activity if given before tumour growth; it was ineffective if given after tumour growth. This study showed that metastatic spread was inversely related to phagocytic activity. The antimetastatic effect of C. parvum appears to be mediated through macrophages in concert with a subpopulation of T lymphocytes, which were considered to be necessary in the sensitization arm of the response as opposed to the effector arm of this response.


Possible transformation of nasopharyngeal epithelial cells in culture with Epstein-Barr virus from B95-8 cells. Explants of fresh biopsy specimens from non-neoplastic nasopharyngeal (NP) mucosa, nasopharyngeal carcinoma (NPC), other tumours (OT) of the head and neck and freshly removed tonsils were treated with an Epstein-Barr virus (EBV) preparation from B95-8 cells and cultured. The mainly epitheloid outgrowths from these infected explants were then compared with those from their respective uninfected controls at 14 days. Growth stimulation occurred with a significantly higher frequency, and the degree of stimulation was generally higher with the infected NP explants than those of the similarly infected explants of other origins. Furthermore, after treatment with the virus preparation, several of the outgrowths from the NP explants showed growth characteristics and cellular morphology typical of those of transformed cells. Light microscopy has shown the changed NP cells to have epithelial characteristics. This is now being verified by electron microscopy, which has so far shown the presence of keratin fibrils and desmosomes in one specimen examined. They are also being examined for the presence of EBV-DNA and EBNA, and other features of transformation, including malignant tendency, by passage through athymic nude mice.


Ultrastructural changes of intercellular junctions in rat ascites hepatoma cells with calcium depletion. To analyse the effect of ethylenediamine tetraacetate (EDTA) on tumour cell adhesiveness, fine structure of intercellular junctions of rat ascites hepatoma cells AH136B and AH7974 (both forming cell islands in vivo) was first compared. The close contact of the apical portion of both cell islands was composed of tight junctions with a narrow gap. The close contact of the inner portion of AH136B cell islands was largely by simple apposition, while that of AH7974 cell islands had many intermediate junctions and desmosomes. Treatment with EDTA (2 mM) induced morphological alteration of simple apposition, intermediate junctions and desmosomes, but tight junctions remained intact. The effect of EDTA on such junctional complexes seemed to be partially reversible on readministration of Ca ions. Changes in desmosomes, as confirmed on AH7974 cells, were initiated by disappearance of the central disc of electron-dense materials, followed by marked opening of intercellular space and disappearance of endoplasmic laminar plaque. These results suggest that Ca ions may be concerned with maintaining the integrity of junctional complexes other than tight junctions.


Experimental infection of rabbits with the virus of infectious bovine rhinotracheitis. Adult and newborn rabbits were infected with the LA strain of infectious bovine rhinotracheitis virus. Adult rabbits developed only mild, self-limiting conjunctivitis, but had microscopic inflammatory foci in the liver and adrenal glands. Newborn rabbits developed a severe, sometimes fatal, generalized infection with focal and diffuse necrosis of the liver and adrenal glands.


Interspecies homology of RNA tumor virus proteins. We report the application of a highly sensitive column chromatographic technique to the comparison of tryptic peptide maps of some RNA tumor virus proteins. By combining microbore ion-exchange chromatography with a sensitive fluorescent assay using o-phthalaldehyde, we obtained high-resolution peptide maps starting with only microgram amounts of protein. Our discovery of coincident peptides from the 15,000 and 30,000 molecular weight proteins from murine and feline leukemia viruses supports serological evidence for interspecies antigenic determinants; coincident peptides were also found for the 10,000 molecular weight proteins from these viruses, although immunochemical data did not reveal interspecies determinants. The relatively large number of coeluting peptides found in the 15,000 and 10,000 molecular weight proteins is strong evidence for the existence of homology.


Studies on conformation of soluble and immobilized enzymes using differential scanning calorimetry. 1. Thermal stability of nicotinamide adenine dinucleotide dependent dehydrogenases. The technique of differential scanning calorimetry (DSC) has been applied to the study of temperature-induced irreversible denturation and thus to the heat stability of soluble and Sepharose-bound liver alcohol dehydrogenase (LADH, EC 1.1.1.1) and lactate dehydrogenase (LDH, EC 1.1.1.27) in the presence of various coenzymes or coenzyme fragments. The transition temperature (Ttr) of 82.5 degrees C obtained for soluble LADH was increased by 12.5 degrees C in the presence of a saturating concentration of NACH. In the presence of NAD+, Ttr increased by 8.5 degrees C, whereas ADP-ribose and AMP caused an increase in Ttr of only 2 and 1 degree C, respectively. The Ttr of 85.5 degrees C obtained for Sepharose-bound LADH was increased by about 12 degrees C after the addition of free NADH. However, when the enzyme was immobilized simultaneously with a NADH analogue (which also binds to the matrix), a broad endotherm with a Ttr of 91.5 degrees C was obtained, indicating the presence of immobilized enzyme molecules both with, and without, associated NADH. Corresponding increases in heat stability were observed for LDH in solution in the presence of NADH, NAD+, and AMP, leading to increases in Ttr from 72 to 79.5 and 74 and 73 degrees C, respectively. The addition of pyruvate and NAD+ to the enzyme to form an abortive ternary complex led to the same stabilization as that observed with NADH, attendant with a large increase in the enthalpy of transition, deltaHtr. In these studies the technique of DSC was utilized because it is applicable both to soluble and immobilized enzymes and (1) provides rapid information about Ttr and thus thermal stability of enzymes, (2) different energetic states of an enzyme molecule can be identified, and (3) an overall picture of the thermal process is rapidly obtained.


Iodinated neurohypophyseal hormones as potential ligands for receptor binding and intermediates in synthesis of tritiated hormones. [3-Iodo-Tyr2]oxytocin (MIOT), [3,5-diiodo-Tyr2]oxytocin (DIOT), [3-iodo-Tyr2,Lys8]vasopressin (MILVP), [3,5-diiodo-Tyr2,Lys8]vasopressin (DILVP), [3-iodo-Tyr2,Arg8]vasopressin (MIAVP), and [3,5-diiodo-Tyr2,Arg8]vasopressin (DIAVP) were synthesized by iodination of the respective hormones, pruified, and characterized. All the monoiodo hormones had to be freshly prepared prior to bioassays, since on storage they gave rise to hormonal-like biological activity. The biological activities of these iodo analogues were measured in an adenylate cyclase assay employing neurohypophyseal hormone (NHH) sensitive bovine renal medullary membranes, and/or the rat oxytocic assay. In the cyclase assay, DIOT, DILVP, and DIAVP were inactive as agonists or antagonists. MIOT shows no agonistic activity in the renal cyclase system and uterus, but is a weak reversible inhibitor of oxytocin (OT) in both systems. When MIOT (10(-4) M) was preincubated with renal membranes for 10 min at 37 degrees C before addition of OT, it behaved as a noncompetitive inhibitor of NHH-stimulated adenylate cyclase. MILVP and MIAVP appear to be partial agonists with Km (half maximal response) 3 X 10(-6) and 3 X 10(-7) M, respectively, as determined in the cyclase assay. Upon preincubation with renal medullary membranes, MILVP (10(-6) M) behaves as a more potent noncompetitive inhibitor of OT than MIOT. Accordingly, iodo derivatives of NHH do not exhibit sufficient affinity to serve an specific ligands to measure OT, LVP, or AVP receptors in the uterus and kidney. Study of the specificity of inhibition produced by MIOT revealed that this analogue does not act selectively upon NHH receptors. Thus, MIOT modified adenylate cyclase systems which do not have NHH receptors, e.g., the PTH-sensitive adenylate cyclase in bovine renal cortex and the glucagon-sensitive adenylate cyclase in rat liver. DIOT, DILVP, and DIAVP were subjected to catalytic tritiation (employing carrier free tritium) and were converted to [3H]OT (25, 31, and 25 Ci/mmol), [3H]LVP (26 and 23 Ci/mmol), and [3H]AVP (17 Ci/mmol), respectively. These tritiated ligands have been successfully used to measure NHH receptor sites both in kidney and uterine membranes as described in other studies.


Physical properties of the dimyristoylphosphatidylcholine vesicle and of complexes formed by its interaction with apolipoprotein C-III. The structure of a single bilayer vesicle of dimyristoylphosphatidylcholine has been characterized by sedimentation, densimetry, and light-scattering measurements. The molecular weight, partial specific volume, Stokes radius, and degree by hydration were found to be 2.68 X 10(6), 0.972 cm3/g, 125 A, and 0.86 g/g, respectively. From these quantities, a spherically symmetrical model has been derived that features a phospholipid bilayer 35.5 A thick and a hydration shell 9.3 A thick. This particle was shown to bind apolipoprotein C-III (apoC-III) up to 0.08 g/g without loss of its original vesicular structure. At protein-lipid ratios in excess of 0.08 g/g, sedimentation, gel chromatography, and light-scattering measurement indicated a dramatic decrease in Stokes radius and molecular weight. The sedimentation data showed these parameters to become constant at protein-lipid ratios in excess of 0.25 g/g. In this region, the Stokes radius and molecular weight were found to be approximately 80 A and 442 000, respectively. Within the constraints of these values and other data, several models for this complex are discussed.


Characterization of baboon plasma high-density lipoproteins and of their major apoproteins. Baboon high-density lipoproteins (HDL) were isolated by preparative ultracentrifugation between d = 1.063 and 1.215 g/mL. The HDL contains 48.8% protein and a lipid distribution similar to human HDL. The phospholipid distribution shows a low sphingomyelin value (5.9%), and the fatty acid composition of HDL is comparable to the human data except for the 18:1/18:2 ratio as a result of a higher 18:1 content in the CE and a lower 18:2 concentration in the PL. The major HDL apoproteins isolated on diethylaminoethyl-cellulose had a mobility on sodium dodecyl sulfate--polyacrylamide gel electrophoresis and a molecular weight and an amino acid composition similar to human apoA-I. However, the amino acid sequence of the first 30 residues of baboon apoA-I differed from the human apoprotein in residues 15 and 21. Treatment of apoA-I with carboxypeptidase A indicated a carboxyl-terminal sequence of Leu-Ser-Thr-Gln. Baboon apoHDL contained monomeric apoA-II with the mobility of monomeric human apoA-II and a molecular weight of 8500. The amino acid composition differed from the human apoA-II by the presence of arginine and by the absence of half-cystine and isoleucine. The circular dichroic spectra of apoA-I and apoA-II demonstrated a higher helicity compared to the human apoproteins. Recombination studies by microcalorimetry of apoHDL with dimyristoylphosphatidylcholine (DMPC) indicated similarities in the thermodynamic binding properties of the HDL apoproteins from man and baboon. The maximal-binding enthalpies of DMPC to apoHDL, apoA-I, and apoA-II were lower for the baboon than for the human apoprotein.


Bovine liver fructokinase: purification and kinetic properties. Fructokinase from beef liver has been purified 2300-fold by acid and heat treatment, ammonium sulfate fractionation, and chromatography on Sephadex G-100, DEAE- and CM-cellulose. The purified enzyme is homogeneous by all criteria examined, has a molecular weight of 56 000, and is a dimer of equal molecular weight subunits. The isoelectric point is 5.7. The Michaelis constant for activation by K+ is 15 mM, and the enzyme is also activated by Na+, Rb+, Cs+, NH4+, and TL+. The kinetic mechanism has been determined at pH 7.0, 25 degrees C. The initial velocity, product, and dead-end inhibition patterns for CrATP, CrADP, and 1-deoxy-D-fructose are consistent with a random kinetic mechanism with the formation of two dead-end complexes. Substrates for fructokinase include: D-fructose, L-sorbose, D-tagatose, D-psicose, D-xylulose, L-ribulose, D-sedoheptulose, L-galactoheptulose, D-mannoheptulose, 5-keto-D-fructose, D-ribose, 2,5-anhydro-D-mannitol, 2,5-anhydro-D-glucitol, 2,5-anhydro-D-mannose, 2,5-anhydro-D-lyxito.l, and D-ribono-gamma-lactone. 5-Thio-D-fructose was not a substate, but was a competitive inhibitor vs. D-fructose. Thus the minimum molecular for substrate activity seems to be (2R)-2-hydroxy-methyl-3,4-dihydroxytetrahydrofuran. The configuration of the substituents at carbons 3, 4, and 5 appears not to be critical, but the hydroxymethyl group must have the configuration corresponding to beta-D-(or alpha-L-) keto sugars. The anomeric hydroxyl on carbon 2 is not required (although it contributes to binding), and a wide variety of groups may be present at carbon 5.


Enzymic formation of glycolate in Chromatium. Role of superoxide radical in a transketolase-type mechanism. Chromatophores prepared from Chromatium exhibit a light-dependent O2 uptake in the presence of reduced 2,6-dichlorophenolindophenol, the maximum rate observed being 10.8 micronmol (mg of Bchl)-1 h-1 (air-saturated condition). As it was found that the uptake of O2 was markedly inhibited by superoxide dismutase, it is suggested that molecular oxygen is subject to light-dependent monovalent reduction, resulting in the formation of the superoxide anion radical (O2-). By coupling baker's yeast transketolase with illuminated chromatophore preparations, it was demonstrated that [U-14C]-fructose 6-phosphate (6-P) is oxidatively split to produce glycolate, and that the reaction was markedly inhibited by superoxide dismutase and less strongly by catalase. A coupled system containing yeast transketolase and xanthine plus xanthine oxidase showed a similar oxidative formation of glycolate from [U-14C] fructose 6-P. It is thus suggested that photogenerated O2- serves as an oxidant in the transketolase-catalyzed formation of glycolate from the alpha, beta-dihydroxyethyl (C2) thiamine pyrophosphate complex, whereas H2O2 is not an efficient oxidant. The rate of glycolate formation in vitro utilizing O2- does not account for the in vivo rate of glycolate photosynthesis in Chromatium cells exposed to an O2 atmosphere (10 micronmol (mg of Bchl)-1 h-1). However, the enhancement of glycolate formation by the autoxidizable electron acceptor methyl viologen in Chromatium cells in O2, as well as the strong suppression by 1,2-dihydroxybenzene-3,5-disulfonic acid (Tiron), an O2- scavenger, suggest that O2- is involved in the light-dependent formation of glycolate in vivo.


Isolation and characterization of nonhistone chromosomal protein C-14 which stimulates RNA synthesis. The nonhistone chromatin protein, C-14, was extracted from chromatin of Novikoff hepatoma ascites cells and isolated in high purity as shown by its migration as a single dense spot on two-dimensional polyacrylamide gels. Its mobility on sodium dodecyl sulfate gels is consistent with a molecular weight of approximately 70 000. The amino acid composition shows that protein C-14 has an acidic:basic amino acid ratio of 1.8. Its amino terminal amino acid is lysine. Protein C-14 stimulated the incorporation of [3H]UMP into RNA by approximately 30% when added to naked DNA and homologous RNA polymerase I. A 30% stimulation of [3H]UMP incorporation into RNA was also found when protein C-14 was added to an E. coli RNA polymerase system containing either E. coli or Novikoff hepatoma DNA.


Substrate form of D-frutose 1,6-bisphosphate utilized by fructose 1,6-bisphosphatase. Rapid quench kinetic experiments on fructose 1,6-bisphosphatase demonstrate a stereospecificity for the alpha anomer of fructose 1,6-bisphosphate relative to the beta configuration. The beta anomer is only utilized after mutarotation to the alpha form in a process that is not enzyme catalyzed. Studies employing analogues of the acyclic keto configuration indicate that the keto form is utilized at a rate less than 5% that of the alpha anomer, a finding also confirmed by computer simulation of the rapid quench data. Chemical trapping experiments of the keto analogue, xylulose 1,5-bisphosphate, and the normal substrate suggest that interconversion of the acyclic and anomeric configurations is retarded by their binding to the enzyme. A hypothesis is advanced attributing substrate inhibition of fructose 1,6-bisphosphatase to possible binding of the keto species.


Binding of CO to mutant alpha chains of hemoglobin M Iwate; evidence for distal imidazole ligation. The optical contribution of the beta chains to the spectrum of hemoglobin M Iwate (alpha87his leads to tyr)2beta2a was subtracted with the aid of a computer so that the spectrum of ferric alpha chains was obtained. Tyrosinate binding to the heme is suggested from spectral resemblance to ferric heme phenolate in dimethyl sulfoxide. The slow reduction of the abnormal ferric alpha chains in hemoglobin M Iwate by dithionite was studied spectrophotometrically both in the presence and absence of CO. The rate of reduction was found to be dependent on the state of ligation of the normal beta chains. The CO-ligated form of the reduced alpha chains bears strong spectral resemblance to the CO-ligated form of the reduced beta chains suggesting similar structures for the heme-ligand complex. A model compound with similar optical properties to the CO-ligated protein can be prepared in dimethyl sulfoxide from hemin chloride, imidazole, and CO using chromous acetate as the heme reductant. Substitution of phenolate for imidazole produces a spectral entity so different from that observed in the protein as to rule out tyrosinate ligation to ferrous heme of the alpha chains when CO is bound.


Plasma membranes from cardiac cells in culture. Enzymatic radio-iodination, evaluation of preparation and properties of the sarcolema. Plasma membranes from heart (sarcolemma) were prepared by the method of Kidwai, A.M. (1975) Methods in Enzymology (Fleischer, S. and Packer, L., eds.), Vol XXXIA, pp. 134--144, Academic Press, New York). On many occasions the sarcolemmal fraction identified by the enzyme markers such as (Na+ + K+)-ATPase banded at heavier densities (d greater than 1.25 g/ml) than expected for plasma membrane (d less than 1.15 g/ml). Radio-iodination of the membrane was added as an independent marker and conditions for the reproducible preparation of the sarcolemma were studied. Cultured heart cells were enzymatically iodinated under conditions which did not affect viability and labeled primarily the sarcolemma. The distribution of radioactivity in homogenates of cultured cells on the density gradient corresponded to that of the enzymes' activity. The best sarcolemma preparation was obtained with 0.3 M KCl extraction of heart homogenates in the presence of 0.05 M pyrophosphate, especially if the salt was also present during the fractionation by density gradient centrifugation. Alterations in the density were also observed with erythrocytes and cultured liver cells' plasma membrane. The data suggests a meta-stable state of the plasma membranes due to handling or storage which could cause alterations of some of their physical properties (e.g. density).


ESR and optical absorption evidence for free radical involvement in the photosensitizing action of furocoumarin derivatives and for their singlet oxygen production. Frozen aqueous solutions of thymine and its derivatives were irradiated with visible light (lambda greater than 320 nm) in the presence of various furocoumarins. ESR analysis revealed the induction of hydrogen adduct free radicals at C-6 position of thymine, only with those furocoumarin derivatives which show a skin-photosensitizing ability. It has been shown, moreover, that the photocycloaddition of psoralen to thymine, which is responsible for the biological effects of this dye, is inhibited when the induction of free radicals in thymine moiety has been prevented by electron scavengers. It is suggested that the free radicals observed could be involved in the biological photosensitization. The mechanism of free radical generation and singlet oxygen production by furoccoumarins were also investigated.


Specific effects of spermine on ouabain-sensitive and potassium-dependent phosphatase activity of kidney plasma membranes. Specificity of the potassium sites. Specific inhibition of ouabain-sensitive and K+-dependent p-nitrophenyl-phosphatase activity of rabbit kidney plasma membranes by spermine (N,N'-bis(3-aminopropyl)-1,4-butanediamine) was characterized kinetically. 1. Inhibition by spermine was competitive with K+. The Ki for spermine was 31 micronM in the presence of 1 mM Mg2+. 2. Excess Mg2+ inhibited the ouabain-sensitive phosphatase activity in competition with K+. The Ki for Mg2+ was 2.6 mM. 3. Increasing Mg2+ concentrations reduced the spermine inhibition. This could be observed at Mg2+ concentrations higher than that of K+. 4. In the absence of inhibition by Mg2+, spermine was noncompetitive with Mg2+ which was essential for the ouabain-sensitive phosphatase activity. This could be observed at Mg2+ concentrations lower than that of K+. 5. Although Ca2+ was a strong inhibitor of the ouabain-sensitive phosphatase activity in the presence of K+, it produced a small stimulation of the activity in the absence of K+. Approximately 0.1 mM Ca2+ gave the maximum stimulation. 6. The observed Ca2+- and Mg2+-dependent phosphatase activity was inhibited strongly by ouabain and by spermine. The half-maximal inhibition concentrations of ouabain and spermine were 0.1 and 63 micronM, respectively. It is likely that Mg2+, Ca2+ and spermine bind to the same site as does K+.


Lipoamide dehyrogenase immobilized on porous glass. Lipoamide dehydrogenase (NADH:lipoamide oxidoreductase, EC 1.6.4.3) isolate from pig heart and Escherichia coli was covalently coupled by both diazonium and amide bonds to controlled pore glass beads (96% silica). When the enzyme was immobilized in the presence of NAD+, the enzyme no longer exhibited its normal requirement for NAD+ for full activity. If the immobilized enzyme was then treated with NADase, the requirement for NAD+ was restored. Enzyme immobilized in the absence of NAD+ exhibited normal NAD+ dependence both prior to an after NADase treatment. These results are discussed in terms of co-immobilization of NAD+ at or near the allosteric site of the enzyme.


Radial diffusion assay of tissue collagenase and its application in evaluation of collagenase inhibitors. A radial diffusion assay for tissue collagenase (EC 3.4.24.3) has been devised which is simple, sensitive and capable of application to large numbers of samples. The assay employs an agarose matrix containing solubilized lathyritic rat skin collagen as substrate. Fibril formation is induced for 2 h at 37 degrees C subsequent to 41 h digestion at 28 degrees C. The procedure results in sharply defined zones of lysis which may be measured directly or after photography. The characteristics of the procedure are otherwise similar to those reported for other radial diffusion assays. The new method was used to examine the action of 10 compounds which were known or potential inhibitors of tadpole collagenase. The concentration of inhibitor required to produce 50% inhibition is reported for the following compounds: alpha2-macroglobulin, 142 microng/ml; N-acetylcysteine, greater than or equal to 100 mM; cysteine, 8.7 mM; EDTA, 0.46 mM; histidine, greater than or equal to 100 mM; 2,3-dimercaptopropanol, 0.5 mM and mercaptoacetic acid, 70 mM. The procedure also has potential for clinical determinations (e.g. tears, synovial fluid) since assay dishes may be prepared in advance and only 15 micronl of sample is required.


An essential arginine residue in the ATP-binding centre of (Na+ + K+)-ATPase. 1. Incubation of purified (Na+ + K+)-ATPase (ATP phosphohydrolase EC 3.6.1.3) from rabbit kidney outer medulla with butanedione in borate buffer leads to reversible inactivation of the (Na+ + K+)-ATPase activity. 2. The reaction shows second-outer kinetics, suggesting that modification of a single amino acid residue is involved in the inactivation of the enzyme. 3. The pH dependence of the reaction and the effect of borate ions strongly suggest that modification of an arginine residue is involved. 4. Replacement of Na+ by K+ in the butanedione medium decreases inactivation. 5. ATP, ADP and adenylyl imido diphosphate, particularly in the presence of trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid to complex Mg2+, protect the enzyme very efficiently against inactivation by butanedione. 6. The (Na+ + Mg2+)-dependent phosphorylation capacity of the enzyme is inhibited in the same degree as the (Na+ + K+)-ATPase activity by butanedione. 7. The K+-stimulated p-nitrophenylphosphatase activity is much less inhibited than the (Na+ + K+)ATPase activity. 8. The ATP stimulation of the K+-stimulated p-nitrophenylphosphatase activity is inhibited by butanedione to the same extent as the (Na+ + K+)-ATPase activity. 9. Modification of sulfhydryl groups with 5,5'-dithiobis(2-nitrobenzoic acid) protects partially against the inactivating effect of butanedione. 10. The results suggest that an arginine residue is present in the nucleotide binding centre of the enzyme.


Altered regulation of cyclic AMP-dependent protein kinase in a mouse lymphoma cell line. The ability of cyclic AMP to inhibit growth, cause cytolysis and induce synthesis of cyclic AMP-phosphodiesterase in S49.1 mouse lymphoma cells is deficient in cells selected on the basis of their resistance to killing by 2 mM dibutyryl cyclic AMP. The properties of the cyclic AMP-dependent protein kinase (ATP:protein phosphotransferase, EC 2.7.1.37) in the cyclic AMP-sensitive (S) and cyclic AMP-resistant (R) lymphoma cells were comparatively studied. The cyclic AMP-dependent protein kinase activity or R cells cytosol exhibits an apparent Ka for activation by cyclic AMP 100-fold greater than that of the enzyme from the parental S cells. The free regulatory and catalytic subunits from both S and R kinase are thermolabile, when associated in the holoenzyme the two subunits are more stable to heat inactivation in R kinase than in S kinase. The increased heat stability of R kinase is observed however only for the enzyme in which the catalytic and cyclic AMP-binding activities are expressed at high cyclic AMP concentrations (10(-5)--10(-4) M), the activities expressed at low cyclic AMP concentrations (10(-9)--10(-6) M) being thermolabile. The regulatory subunit of S kinase can be stabilized against heat inactivation by cyclic AMP binding both at 2-10(-7) and 10(-5) M cyclic AMP concentrations. In contrast, the regulatory subunit-cyclic AMP complex from R kinase is stable to heat inactivation only when formed in the presence of high cyclic AMP concentrations (10(-5)M). The findings indicate that the transition from a cyclic AMP-sensitive to a cyclic AMP-resistant lymphoma cell phenotype is related to a structural alteration in the regulatory subunit of the cyclic AMP-dependent protein kinase which has affected the protein's affinity for cyclic AMP and its interaction with the catalytic subunit.


Purification and molecular properties of the AMP-activated pyruvate kinase from Escherichia coli. The AMP-activated pyruvate kinase (ATP:pyruvate 2-O-phosphotransferase, EC 2.7.1.40) from Escherichia coli has been purified 200 times through a three-step procedure which gives a homogeneous preparation with a specific activity of 110. The enzyme appears to be a tetramer of molecular weight 190 000. Subunits (molecular weight 51 000) show a single amino-terminal amino acid (serine) and appear as a single band in polyacrylamide gel electrophoresis in sodium dodecyl sulphate. The enzyme crystallizes in conditions of reduced dielectric constant of the solvent in the pH range 6.5-7.5. Kinetic and regulatory properties of the purified enzyme are similar to those described for crude preparations of the enzyme.


The role of bound thiamine pyrophosphate in the synthesis of thiamine triphosphate in rat liver. Thiamine pyrophosphate-ATP phosphoryltransferase, the enzyme that catalyzes the synthesis of thiamine triphosphate, has been found in the supernatant fraction of rat liver. The substrate for the enzyme is endogenous, bound thiamine pyrophosphate, since the addition of exogenous thiamine pyrophosphate had no effect. Thus, when a rat liver supernatant was incubated with gamma-labelled [32P]ATP, thiamine [32P]triphosphate was formed whereas the incubation of thiamine [32P]pyrophosphate with ATP did not produce thiamine [32P]triphosphate. The endogenous thiamine pyrophosphate was found to be bound to a high molecular weight protein which comes out in the void volume of Sephadex G-75, and is not dialyzable. The activity that catalyzes the formation of thiamine triphosphate has an optimum pH between 6 and 6.5, a linear time course of thiamine triphosphate synthesis up to 30 min, and is not affected by Ca2+, cyclic GMP and sulfhydryl reagents.


The proteolytic action of bacterial proteases from Clostridium histolyticum on bovine fibrinogen. Bacterial proteases from Clostridium histolyticum bring about the polymerization of fibrinogen into structured fibres, the period being 9 nm when the ionic strength (I)  of the solution is between 0.1 and 0.2. At I=0.3 no polymerization occurs, but the successive actions of bacterial proteases and thrombin induce polymerization, the fibres obtained showing a periodic structure: the major period is 23 nm with two minor striations. The striation pattern is different from that obtained with fibrin which shows a major period of 23 nm with three minor striations. The degree of degradation of fibrinogen monitored by disc gel electrophoresis shows that the Aalpha polypeptide chain (mol. wt.=68 000) is degraded into Aalpha1 (mol. wt.=32 000) and Aalpha2 (mol. wt.=29 000), whilst the mobility of Bbeta increases and gamma remains unchanged; the molecular weight is estimated between 272 000 and 269 000. These results emphasize that the charge distribution differences which are compatible with the loss of different portions of the molecule, lead to variations in fibre structures.


[Extraction of deuterated compounds from Chlorella sp. K. grown in D20]. A comparison of the three known methods made it possible to work out methods of complex extraction of deutero amino acids, deutero sugars and deutero vitamins from algae Chlorella sp. K., grown autotrophically in heavy water. An analysis of the isotopic composition of deutero amino acids by PMR indicates that treatment with HCl introduces 1H into side chains of some amino acids by chemical exchange (C2, 6H of tyrosine, C2H of histidine, rho-H of phenylalanin, beta-CH2-aspartate and gamma-CH2-glutamate. These exchanges made the amino acids convenient to be used as the substrates for the growth of heterotrophic selective deuterated organisms as a source of selective deuterated proteins.


[Cooperative properties of D-glyceraldehyde-3-phosphate dehydrogenase]. The structure of the active center of glyceraldehyde-3-phosphate dehydrogenase and the arrangement of subunits in the tetrameric molecule is delineated. The mechanism of cooperative effects in the oligomer is considered, and the involvement of various regions of the active center and of different-subunit contact area in the realization of the cooperative phenomena is discussed. A special attention is paid to the effect of NAD+ bound to one of the subunits of the tetramer on the structure of an adjacent subunit and to the problem of the participation of the coenzyme in the creation of anion-binding sites of the enzyme. The conditions of reversible dissociation of the tetrameric apoenzyme molecule into dimers are depicted, and the role of NAD+ in the organization of the quaternary structure of the dehydrogenase is discussed. The problem of catalytic activity of the dimeric form of the enzyme is argued.


[2 types of reduction of P700+ reaction centers in the leaves of green plants]. It is shown that in the leaves of Hibiscous sp. the ESR signal I rises under weak far red light after its partial inhibition with a short white flash is slower than after adaptation to 710 nm light and short dark exposition. It is suggested that this divergence in the ESR kinetics is due to the fact that only part of reduced intermediates generated by the photosystem 2 under flash is readily available to P700+, whereas other reducing reagents slowly react with the photosystem 1 reaction centres.


Thyroxine effect upon activity of uridine kinase in developing rat cerebellum. Experimental hyperthyroidism in the neonatal rat is known to accelerate cerebellar DNA biosynthesis resulting ultimately in a deficit in cell number at maturity. Because of the know shift to an earlier age in the developmental curve for cerebellar thymidine kinase activity in rats treated with thyroxine, we studied the activity of uridine kinase and DNA biosynthesis during rat cerebellar development under hyperthyroid conditions. Body weight and cerebellar wet weight in treated animals were noted to be significantly decreased below control values on days 4 and 12, respectively. Cerebellar DNA was significantly elevated above control values on days 4 and 6 (132 and 129% of control, respectively). Subsequently, DNA content fell significantly below control values through day 18. Uridine kinase activity was found to be increased significantly above control values at ages 2, 4, and 6 days (maximum 119% of control at age 4 days) following which activity fell significantly below control values by 15 days of age. Uridine kinase activity from both treated and control animals fell only moderately after the time of peak activity between 9 and 15 days of age, although the peak of the developmental curve for the enzyme appeared earlier in the treated animals. The data show a less pronounced early stimulation of cerebellar uridine kinase by thyroxine compared with previously reported thyroxine enhancement of thymidine kinase activity, although both enzymes are affected by thyroxine throughout cerebellar ontogenesis. The study thus provides evidence that uridine kinase is sensitive to hormonal stimulation during early stages of active cerebellar cell division, and that the enzyme may relate most closely in brain to the synthesis of RNA as well as the sustaining of cell function after the most active phase of cellular proliferation. In addition, the study emphasizes the use of enzyme-hormone relationships during development to provide information concerning critical interrelationships between metabolic pathways contributing to nucleic acid biosynthesis.


Behavioral and psychological correlates of a difference in chronic sleep duration. The relationship between chronic differences in sleep duration and waking behaviors was explored by comparing two groups of 10 healthy male university students who regularly slept nocturnally for 7-8 h or for 9.5-10.5 h. Measurements were obtained of sublingual temperature, from a 45 min Wilkinson auditory vigilance task and a mood adjective check list 30 min after awakening in the morning, at midday ind in the early evening following an electroencephalographically recorded night of sleep. In both subject groups body temperature increased from morning to early evening, while misses on the vigilance task correspondingly declined during the day. The average daily level of oral temperature and performance were significantly lower in the 7-8 h (control) group than in the long sleepers. Positive mood states (Cheerful, Energetic, General Activation, High Activation) were significantly  greater and negative affects (Anger-Hostility, Depression) significantly less in the long sleepers. As a result of the mean difference in total sleep time existing between groups control subjects averaged significantly less stage 2, and stage REM sleep. It was postulated that there were behavioral deficits in the control group attirbutable either to selective sleep stage deprivation or to the general consequence of reduced sleep per se or to both of these factors.


EEG theta waves and psychological phenomena: a review and analysis. In this paper, studies which have explored the relation between EEG theta waves and psychological phenomena in normal human subjects are reviewed. It is noted that increases in theta activity occur in conjunction with several kinds of psychological processes. The importance of ocnsidering properties of theta activity, such as amplitude, rhythmicity and scalp topography when analyzing the relation between theta and psychological processes is emphasized. Although there is some evidence for a relationship between theta and psychological processes, it is concluded that the degree to which properties of theta activity are systematically related to specific psychological processes is not yet known.


Children sea-blue histiocytosis (2 cases) compared with phospholipidosis induced by 4-4' DET (4-4' p (diethylamino-2-ethoxy phenyl) 3-4 hexane) in rat. Two cases of children with liver and spleen enlargement are reported. Sea-blue histiocytes and Pick cells were found in both cases in liver, spleen, bone marrow and blood. Further more, lysobisphodphatidic acids were identified in phospholipid analysis of liver  biopsies and cultived liver cells. Absence of neurologic involvement at 14 and 18 years fo age suggest a Crocker type C of Niemann-Pick disease, i.e. a not yet well defined entity. Resemblance of these morphological and biochemical abnormalities with certain cases of drug poisoning (especially the well-known intoxication by 4-4' DET)  is discussed on the basis of results from experimental studies with this drug in the rat.


Early syncytium formation induced by bovine leukemia virus in mixed cultures. Early syncytium formation was obtained in mixed cultures in the presence of BLV (Bovine Leukemia Virus). The phenomenon is specifically dependent upon the presence of BLV.


[Quantitative analysis of the process of stimulation frequency transformation in the neuromuscular apparatus]. Dependence of parameters of the neuromuscular transmission on the stimulation frequency was determined. This dependence was analyzed as applied to the processes of synapsis "fatigue" occurring in prolonged rhythmic muscle stimulation. The suggested mathematical model and the method of statistical analysis of the EMG of the sum total muscle responses following from it offered a possibility of determination by the experimental data of the approximate parameters giving a quanititative expression of the frequency properties of the neuromuscular transmission.


[Formation of cyclic adenosine-3',5'-monophosphate in phagocytosis]. The content of cAMP in the phagocytizing macrophages increased, especially in the phagocytosis of live microbes. cAMP formed in phagocytosis was determined in the incubation medium, whereas in the cells its content remained practically unchanged. In the administration of E. coli 055 to the germ-free guinea pigs the concentration of cAMP was found to be increased in the mucosa cells of the small intestine and in the blood serum; this fact indicated that adenylcyclase system participated in the reactions of the interrelations of the microorganisms with the epithelial cells of the smal intestiine.


[Spontaneous fluctuations in the activity of rabbit skeletal muscle NAD-kinase]. Spontaneous fluctuations in the time of the activity of the 280-300-fold purified NAD-kinase preparation from rabbit skeletal muscle following its dilution are described. Defrosted but undiluted enzyme preparation failed to exhibit any fluctuations in its activity.


[Changes in the kinin system components and structure of the myocardium under the influence of antikinin preparations in allergic myocarditis in rabbits]. Experimental allergic myocarditis was induced in rabbits; a study was made of the effect on the kininogen content and on the activity of kininase in the myocardium, and on its morphological structure produced by anginine, contrical, aspirin, indometacine and epsilon-aminocapronic acid. The former two preparations were much more potent (in comparison with the others) in inhibiting the activation of the myocardial kinin system and promoted restoration of the morphological structure of the myocardium. Development of an allergic inflammation in the myocardium could be arrested or decreased by inhibition of the kinin system of the plasma and myocardium by specific agents.


Antagonism by PGF2alpha of the vasodilation induced by PGE1, PGE2 and PGA1 in the canine uterus. Prostaglandins appear to play an important role in a number of reproductive processes. The present study was designed to determine if the vasodilator action of prostaglandins of the A and E series on uterine vascular smooth muscle was mediated via a receptor and if PGF2alpha could compete for this same receptor. The data demonstrate that PGF2alpha, which by itself has no uterine vascular effect, can produce a parallel shift in the dose-response curves for certain vasodilator prostaglandins. This action, which suggests competitive antagonism, may well play a role in regulating blood flow in the nonpregnant uterus.


Current status of population kinetics in gliomas. One must know tumor cell kinetics in order to devise a rational drug regimen for gliomas. With tritiated thymidine, the Labelling Index of astrocytomas is less than 1 per 100; of glioblastomas from 5-10 per 100. The duration of S phase is fairly constant, ranging from 7 to 10 hours. Double radioautography reveals a cell cycle time of 2 to 3 days and a Growth Fraction of 10 to 30 per 100 in glioblastomas. Calculations based on volume of tumor at recurrence after a gross total resection, analysis of primer dependent D.N.A. polymerase (P.D.P.), and analysis of cells by Flow Microfluorometer, all confirm these approximate figures. Thus most of the cells of a glioma are not sensitive to a cell-cycle phase specific drug. Such an agent, if given, should be administered over a 2 to 3 day period, in order to affect as many cells as possible. The most important part of a drug regimen should be an agent which attacks non-proliferating as well as proliferating cells, such as an alkylating agent. The effects of various drugs and schedules can be examined by animal models with the technqiue of colony forming efficiency.


[Treatment of inoperable glioblastomas using a combination of adriamycin, VM 26 and CCNU]. 46 non resectable malignant gliomas adult patients were treated with a combination protocol of chemotherapy with administration of adriamycin, VM 26 and CCNU. The tolerance to this treatment was good except for the delayed hematological toxicity related to cumulative doses of CCNU, which limited after 5 courses of chemotherapy the possibility of further treatment with the same combination. As far as the neurological responses are concerned, 66 per 100 patients were good responders, but we found a poor relation (50%) between the degree of clinical response and regression of tumour volume in brain scan. The median survival of this group of patients was 9 months and 11 patients/46 are still alive in good state 14 months after they entered into this trial. According to these data we discuss the introduction of chemotherapy at the early stage of the disease.


[Concentrated irradiation of malignant astrocytoma and glioblastoma]. Since 1965, 118 glioblastoma and 18 malignant astrocytomas of the adult have been treated by concentrated irradiation after a more or less complete surgical excision of the tumor. Three types of irradiation have been used; at present 3 600 rads whole brain irradiation are delivered in 2 series of 1 800 rads over 3 days 20 to 30 spaced a part. All patients receive ACTH and the tolerance has been excellent. The results of this rapid palliative therapy are quite comparable to those of more classical irradiation. Because of the very short survival of these patients, it would appear advantageous to treat them in as short a time interval as possible.


[Intramammary spread of breast cancer].

[Intramammary spread of breast cancer]. At the conclusion of this study based on a careful histological analysis of 454 mastectomy specimens carried out at the Institut Gustave-Roussy between 1966 and 1971 it appears that 39 per cent of tumours which are neither central nor deep to the nipple have microscopic foci outside the main focus of tumour of which 24 per cent are at the level of the single nipple. The importance of these extensions is bound up with the size and histological type of the adenocarcinoma: pure intraduct and infiltrating lobular forms are accompanied frequently by microscopic intramammary foci at a distance (89% and 72%) the opposite of findings in the case of well differentiated and medullary forms. On the other hand no connection between the Scarff and Bloom grading is apparent except where the nipple only is involved. Finally there is a strong relationship between intramammary dissemination and regional lymph node metastases.


Enteroviruses other than poliovirus.

Enteroviruses other than poliovirus. The role of enteroviruses in certain specific disease--cardiac disease, nephritis, diabetes, and hemorrhagic conjunctivitis--is examined. It has now been well documented that Coxsackievirus B (types CB1 through CB5 but not CB6) is the main pathogen involved in various clinical forms of viral heart disease. Coxsackievirus A (CA4 and CA16) and echovirus (types 9 and 22) may also be associated with viral heart disease. In regard to the etiologic role of enteroviruses in nephritis, pancreatitis, and diabetes, again CBV, especially CB3 and CB4, has been suspected, but the data are controversial and further studies are needed. Hemorrhagic conjunctivitis, a newly observed clinical entity, is caused by enterovirus 70. It has spread to four continents (not including the Americas) in a pandemic fashion since 1969 and is now one of the common eye infections in these areas. The virus has some neurovirulence, and motor paralysis is known to occur as a complication; hence it should be carefully watched in the future.


Blocked beds.

Blocked beds. In a cross-sectional survey of 325 surgical and orthopaedic beds 43 (16%) of the 265 occupied beds were filled by patients who had no medical need to be in an acute ward. They had been in hospital for a median time of 40 weeks up to the survey date. Of the 43 patients, 11 were awaiting transfer to a geriatric ward; 13 to community residential care; and seven to their homes. There was no plan for discharge or transfer for the remaining 12 (28%). Those "at risk" of becoming long-stay patients for social reasons on these wards were women, over 75, living alone or with one relative, who had been admitted to hospital in emergency with a fractured femur, head injury, or other trauma. Action necessary to reduce the number of social long-stay patients includes (a) changing attitudes to the solving of social case problems; (b) revising procedures of assessment and planning of future care; (c) improving teamwork and record keeping within the hospital and the community services; (d) providing a better balance of acute, medium and long-stay hospital beds; and (e) putting more resources into rehabilitation.


The ultrastructural characteristics of the abnormal cytosomes in Batten-Kufs' disease. Patients with Batten-Kufs' disease may be divided into three groups by electronmicroscopy of their storage deposits. In the first group, those characterized by curvilinear profiles, there is a strong correlation with a particular clinical syndrome, the late infantile form of the disease. In the second group, characterized by finger-print profiles, there is great diversity as to age and type of presentation. This is paralleled by diversity in the deposits. To the third group belongs the infantile form of the disease, as well as rare patients with later onset. Pathological diagnosis can be reliably, conveniently and consistently made from biopsy of skin by electronmicroscopy, and usually from biopsy of skeletal muscle as well.


Isolation of glial cell-enriched and -depleted populations from mouse cerebellum by density gradient centrifugation and electronic cell sorting. Preparative amounts of populations enriched and depleted in glial cells have been isolated from 10-day-old mouse cerebella. Discontinuous density bovine serum albumin (BSA) gradients provide two distinct populations: the one derived from the 10-15% BSA interface is enriched in cyclic nucleotide phosphohydrolase (CNPase) activity, and a percentage of S-100, GFA protein, and NS-1 antigen-positive cells; the other, located as the 15-31% interface, contains a cell population depleted in these compounds. This report also describes the first use of flow microfluorimetry and electronic cell sorting techniques for the analysis and isolation of cell populations derived from the mammalian central nervous system. Glial cell-enriched and -depleted populations obtained by BSA density gradient centrifugation were analyzed for their forward angle light scattering properties and their capacity to bind anti-corpus anti-serum to the cell surface. The glial cell-enriched fraction shows a different frequency distribution of light scattering than the glial-depleted fraction. Anti-corpus callosum antiserum binds preferentially to the glial cell-rich fraction. Cells can be sorted into corpus callosum antigen-positive and -negative cell fractions and recovered with a viability of more than 95% as judged by trypan blue exclusion. Anti-corpus callosum-positive sorted cells are enriched in CNPase activity, S-100, and glial fibrillary acidic proteins.


[Cyto-immunological study of the Pleurodele pituitary gland]. In the pituitary of an Amphibian (Urodela), various cell types were localized by cytoimmunological techniques: corticotrophs located in a rostral zone near the median eminence, ventral orangeophilic cells secreting a prolactin-like hormone and erythrosinophilic cells in a more dorso-caudal situation secreting growth hormone. Alpha-MSH and beta-MSH produceng cell were identified in the intermediate lobe; these cells were also labelled with antibodies against 1-24 ACTH and 17-39 ACTH. These data are in accordance with results obtained after an experimental study of the prolactin cells.