Regulation of sterol synthesis in leukaemic blast cells: a defect resembling familial hypercholesterolaemia. Leucocytes from normal subjects and from patients with acute myeloblastic leukaemia incubated for 9 h in the absence of lipoproteins increased their rate of sterol biosynthesis from [2-14C]acetate threefold. Subsequent addition of complete serum, low density lipoprotein or cholesterol in a non-lipoprotein form suppressed sterol synthesis in leucocytes from normal subjects. In contrast, complete serum or low density lipoprotein totally failed to suppress sterol synthesis in leukaemic blast cells. However, when these cells were incubated with cholesterol in a non-lipoprotein form suppression of sterol synthesis occurred as in normal leucocytes. These results suggest that leukaemic blast cells behave like cells from patients with familial hypercholesterolaemia in showing failure of regulation of sterol synthesis by low density lipoprotein.


Plasma equilin concentrations in an oophorectomized woman following ingestion of conjugated equine oestrogens (Premarin). The plasma equilin concentration was 25.2 nmol/l (6.7 ng/ml) 4 h after an oral dose of Premarin (1.25 mg), and a maximum concentration of 20.5 nmol/l (5.5 ng/ml) was recorded 3 h after a smaller dose (0.625 mg) was given. 24 h later, the plasma equilin was still considerable, being 6.1 nmol/l (1.6 ng/ml) and 3.7 nmol/l (1.2 ng/ml) for the larger and smaller dosages respectively. The findings for equilin are considerably higher than physiological oestrogen concentrations and in addition show that daily therapy with Premarin could lead to cumulation of the steroid.


Prevalence of antibodies to soluble antigen of Epstein-Barr virus-producing cells (P3HR-1) in the sera of hamadryas baboons of the high lymphoma incidence stock. Soluble antigen of P3HR-1 cells (SA-P3HR-1) was identified in indirect double immunodiffusion enhanced with tannic acid using serum of a nasopharyngeal carcinoma patient containing high-titer antibodies to Epstein-Barr virus (EBV) antigens. SA-P3HR-1 was nonidentical to soluble antigen of Raji cells. Human and baboon sera containing antibodies to all the known antigen of EBV and HVP respectively were anti-SA-3HR-1-positive. Human and baboon sera containing antibodies to all the known antigens of EBV and herpesvirus Papio (HVP) were also anti-SA-P3HR-1-negative. Prevalence of anti-SA-P3HR-1 was very high in two groups of the high-lymphoma incidence stock of hamadryas baboons of the Sukhumi monkey colony. 54% (15 of 28) and 38% (13 of 34) of clinically lymphomatous and clinical normal monkeys, respectively, were anti-SA-P3HR-1-positive.. Only 1 of 30 normal baboons studied, living in the forest and having no contacts with the baboons in the main stock of the Sukhumi monkey colony, was anti-SA-P3HR-1-positive (3%).


Increased agglutinability and haemadsorption of Marek's disease HPRS line 1 lymphoblastoic cells after concanavalin A treatment. The HPRS line 1 lymphoblastoid cells derived from an ovarian lymphoma induced by Marek's disease virus in chickens displayed increased agglutinability and haemadsorption when incubated with Con A. The increase was proportional to the concentration of Con A used. The same Con A-untreated cells and normal chicken lymphocytes incubated with Con A exhibited no increase in agglutinability and haemadsorption, however.


[Blood serum cortisol, insulin and somatotropin concentration in children with hypothalamo-hypophyseal insufficiency]. In 22 patients with restricted growth the consecutive stimulation with arginine and insuline was applied for the diagnosis of somatotropic insufficiency. 13 children showed GH insufficiency. Simultaneously cortisol level during hypoglycemia for ACTH reserve and insulin level after arginine application were estimated.


Neuropsychological dysfunction in acute viral hepatitis. Neuropsychological dysfunction may precede, occur simultaneously with, or follow the icteric phase of viral hepatitis. Encephalitic and meningitic involvement appears to precede the onset of clinical hepatitis while the tremor syndrome usually follows the onset of illness. The Guillain-Barré syndrome, peripheral nerve disorders, and behavioral abnormalities may occur at any stage of the disease. Neuropsychological alterations noted during the prodrome of viral hepatitis usually clear without sequelae but abnormalities which follow the onset of jaundice may persist during the convalescent phase for prolonged periods. Neither occurrence, type, nor severity of neuropsychological dysfunction appear to be related to specific hepatitis viruses since both type B and non-B infections have been implicated. Furthermore, neuropsychological dysfunction does not appear to be related to the severity of the hepatitis as assessed by clinical or histological features. Although direct viral invasion of nervous tissue and immune complex-mediated damage have been postulated, the mechanism(s) of neuropsychological abnormalities remains to be determined.


Tumor cell aggregation and mode of cancer spread in linitis plastica type of gastric carcinoma. Spread of tumor cells in linitis plastica type of gastric carcinoma was studied in relation to features of tumor cell aggregation which were classified into two types: free-cell type and small nest type. Free-cell type was accompanied by a larger amount of fibrous stroma in submucosal layers than in small nest type. In the gastric wall lymphatics and regional lymph nodes, small nest types was more prominent than the free-cell type. Lymphatic permeation was noted predominantly in the submucosa. In the mucosa, both free-cell and small nest types were frequently present. In carcinoma with free-cell type predominance, diffuse infiltration with infrequent lymphogenous spread was noted, while in carcinoma with small nest type predominance, spread via the lymphogenous route was observed more frequently. The mode of cancer spread in this type of carcinoma appeared to be affected by the type of tumor cell aggregation dominant in the mucosa.


Histopathological studies on renal tubular cell tumors in rats treated with N-ethyl-N-hydroxyethylnitrosamine. Administration of a diet containing 0.2% N-ethyl-N-hydroxyethylnitrosamine (EHEN) for 2 weeks induced hepatocellular carcinoma in 3 and renal tubular cell tumors in 7 of 9 Wistar rats. Diet containing 0.1% EHEN induced renal tubular cell tumors in 6 of 10 rats, but no hepatocellular carcinomas. Both diets induced atypical cell population of tubules in the kidney, but no nephroblastoma.


Sensitive and rapid method for determination of superoxide-generating activity of blood monocytes and its use as a probe for monocyte function in cancer patients. The superoxide anion-generating capacity of human blood monocytes was measured by a sensitive and rapid method established by taking advantage of the fact that the generation of superoxide anions by monocytes was markedly enhanced by the combined stimulation of the cells with cytochalasin-E and wheat germ agglutinin. The activity was expressed by the initial rate of cytochrome c reduction after the addition of wheat germ agglutinin. The rate obtained with normal human monocytes was 0.73 +/- 0.19 nmol/min/10(5) monocytes (mean +/- SD, n = 10). Because of its sensitivity, the method required only 10(5) monocytes and can be used to follow the monocyte function in various patients. Preliminary data obtained with 15 cases of advanced cancer patients (0.29 +/- 0.10 nmol/min/10(5) monocytes) suggested a possible decrease of the superoxide-generating activity, at least in some state of cancer patients. It appears that measurement of superoxide-generating activity will be meaningful to monitor monocyte function.


A mutator affecting the region of the iso-1-cytochrome c gene in yeast. The mutator gene DEL1 in the yeast Saccharomyces cerevisiae causes a high rate of formation of multisite mutations that encompass the following three adjacent genes: CYC1, which determines the structure of iso-1-cytochrome c; RAD7, which controls UV sensitivity; and OSM1, which controls osomotic sensitivity. The simplest hypothesis is that these multisite mutations are deletions, although it has not been excluded that they may involve other types of gross chromosomal aberrations. In contrast, normal strains do not produce such multisite mutations even after mutagenic treatments. The multisite mutations arise at a rate of approximately 10(-5) to 10(-6) per cell per division in DEL1 strains, which is much higher than rates observed for mutation of genes in normal strains. For example, normal strains produce all types of cyc1 mutants at a low rate of approximately 10(-8) to 10(-9). No evidence for multisite mutations was obtained upon analysis of numerous spontaneous ade1, ade2, met2 and met15 mutants isolated in a DEL1 strain. DEL1 appears to be both cis- and trans-dominant. The location of the DEL1 gene and the lack of effect on other genes suggest that the mutator acts only on a region adjacent to itself.


The BAM H1 restriction site in the bacteriophage T4 chromosome is located in or near gene 8. Bacteriophage T4 cytosine-containing DNA is cleaved at a single site by the restriction endonuclease, Bam H1. The site lies within the late region of the T4 genome, close to, or within, gene 8, one of the structural genes of the phage particle baseplate.


Clustered arg genes on a BamHI segment of the Escherichia coli chromosome. BamHI cleavage of DNAs from transducing phages gamma darg13 (ppc, argECBH, bfe), gamma darg14 (ppc, argECBH) and gamma darg23 (argECBH) yields three purely gamma DNA segments (and, in one case, a fourth), as well as several Excherichia coli-DNA-containing segments. The length (in kilobases, kb) of the segments, determined by electron microscopy and ararose gel electrophoresis is 4.2, 7.5, 8.4, 6.2, 6.9, and 6.4 kb for gamma darg13; 13.0, 7.5, 4.7, 6.2, 6.9, and 6.4 kb for gamma darg 14: and 5.3, 11.0, 4.7, 6.2, 6.9, and 6.4 kb for gamma darg23. Ordering of the segments (in relation to the gamma genetic map and with the direction from left to right corresponding to the clockwise orientation of the E. coli genetic map and to each of the numerical sequences given) reveals, on 26 kb of bacterial DNA, two cleavage sites defining the 7.5-kb segment obtainable from the DNA of either gamam darg13 or gamma darg14. These and analogous findings with argEC and argCB deletion-bearing strains, together with results from heteroduplex experiments, locate argE, argC, argB, and presumably argH on the 7.5-kb segment.


A new host-vector system allowing selection for foreign DNA inserts in bacteriophage lambda gtWES. An improved vector (lambda gtWES.T5-622) for EcoRI fragments has been derived from EK2 vector lambda gtWES.lambdaB' by replacing the lambda B fragment with two identical 1.1 Md fragments from the pre-early region of bacteriophage T5. The new vector has two advantages which facilitate elimination of parental-type recombinants in an in vitro recombination experiment. Firstly, the 1.1 Md insert is too small to be re-inserted into lambda gtWES in a single copy. Secondly the 1.1 Md T5 fragment carries T5 gene A3 which prevents growth of phage retaining this fragment when the Excherichia coli host carries plasmid ColIb. Thus, essentially all plaques are due to phage with donor DNA inserts and are free of T5 DNA fragments. The size usually given as the theoretical minimum size for insertion into the lambda gt series of vectors is 0.66 Md. We have shown that this size is an underestimate and that the lower limit is about 1.6 Md. A precise estimate is difficult since there is strong selection, among phage having small inserts, for those which have acquired additional genetic material by duplication of the lambda DNA.


The inverted terminal repetition of the DNA of weakly oncogenic adenovirus type 7. By use of the chemical modification technique of Maxam and Gilbert (1977), the first 180 base pairs at both termini of the human adenovirus 7 genome have been determined. The results show that adenovirus 7 DNA contains a perfect inverted terminal repetition of 136 base pairs.


Stimulation of the alpha-and beta-adrenergic receptors in human ovarian vasculature in vitro. The effect of fenoterol and norepinephrine was studied on the vascular resistance of in vitro perfused human ovary. It was found that the beta-adrenergic stimulant fenoterol causes vasodilatation, whereas the alpha-adrenergic stimulant norepinephrine elicits vasoconstriction in the arterial vascular bed of the ovary showing the role of adrenergic mechanism in the regulation of ovarian blood flow. It was supposed that therapeutical administration of beta-mimetic drugs leads to the elevation of ovarian blood flow, which may have special importance in the case of corpus luteum insufficiency.


The influence of the non-dialysable sugar-peptide fraction from bovine blood plasma on the amino acid incorporation of BHK-21 normal and RSV transformed cells. The non-dialysable sugar-peptide rich fraction of bovine blood plasma has been studied with regard to its influence on the amino acid incorporation of BHK-21 and BHK/RSV cells in vitro. The results indicate that the Nsp fraction contains a peptide factor(s) causing changes in activity for the protein and DNA synthethizing system in both the cell cultures examined. The biological activity of the factor(s) is regulated by interaction with blood serum components and this fact may play a leading role in its regulatory function.


[Changes in the osmotic fragility of erythrocyte membrane in morphine- and phenobarbital-dependent rats (author's transl)]. This is apparently the first attempt to elucidate the relationship between drug dependence and the osmotic fragility of erythrocyte membrane. The osmotic fragility was measured using a coil planet centrifuge (CPC) system. Utilizing the drug-admixed food (DAF) method, rats were made drug-dependent. The osmotic fragility of morphine-dependent rats was significantly enhanced, compared with that of naive rats. By withdrawing or treating the rats with levallorphan, the osmotic fragility was enhanced more than in the morphine-dependent state. When the morphine-withdrawal rats were again given the morphine-admixed food, the osmotic fragility recovered to the morphine-dependent level. The osmotic fragility of phenobarbital-dependent rats was significantly decreased, compared with that of naive rats. On the contrary, in the phenobarbital-withdrawal rats, the osmotic fragility was significantly enhanced, compared with that of the phenobarbital-dependent rats. With re-treatment of phenobarbital-admixed food, the osmotic fragility was recovered to the levels seen in the phenobarbital-dependent rats. Abstinence signs including weight loss, decrease in food and water intake, adrenal hypertrophy etc., were observed during morphine or phenobarbital withdrawal. The effects of food or water deprivation and application of ACTH on the osmotic fragility were then studied and we found that the osmotic fragility was enhanced with these treatments. These results suggest that enhancement of osmotic fragility during withdrawal of these drugs is partly influenced by these treatments.


Surface markers and functional characteristics of human blood lymphocytes residing in the operational "null" subset. After removal of nylon wool-adherent B and E rosetting T cells from the human blood lymphocyte population, a small fraction remains, often denoted as "null" subset. This fraction contains lymphocytes which are highly efficient in antibody-dependent and non-selective lymphocyte cytotoxicity. These cells possess markers. About half of them carry Fc receptors, and the majority has low avidity E and C3 receptors. Removal of low avidity E receptor-carrying cells did not abolish cytotoxicity to K-562 cells. The majority of cells carry receptors for Epstein-Barr virus but cannot be infected. The cells in mixed lymphocyte culture have a low reactivity. They aare stimulated by PHA and ConA to a similar extent and differ from the cells which readily sediment as E rosettes: these respond stronger to PHA. The "null" fraction may contain stem cells common for both T and B lineages.


Cancer, viruses and environmental factors. The genesis of Burkitt's lymphoma is visualized as proceeding in three steps: I. Primary EBV infection affects the young child, probably at a relatively high multiplicity. It immortalizes a certain number of B-lymphocytes in vivo. II. This is followed by the impact of an environmental promoting agent, perhaps chronic holoendemic malaria, providing a chronic stimulus to the proliferation of the EBV-carrying preneoplastic cells. III. Chromosomally abnormal variants appear in the stimulated tissue by chance. After certain types of changes, particularly the 8 to 14 translocation that leads to the 14q+ marker, the affected B lymphocyte would no longer obey the negative feedback controls that would otherwise restrict its proliferation in vivo.


[Hypothalamo-pituitary-adrenocortical (HPA) function in the morphine dependent rat (author's transl)]. Morphine dependence was induced by ten daily administrations of morphine with gradually increasing dosage (40--160 mg/kg/day s.c.) to male Sprague-Dawley rats, the dosage being divided in 2/day for the first 8 days and 4/day for the last 2 days. Diurnal variation in body weight in morphine dependent rats was the same as in naive rats. Although body temperature in the naive rats showed a diurnal variation with the highest at 3:00 and the lowest at 15:00, in the morphine dependent rats there was no distinct variation. Plasma corticosterone level (PC) in naive rats showed diurnal variation with the peak at 21:00 and the trough at 9:00. PC in morphine dependent rats showed the same diurnal variation as did the naive rats, while such disappeared following pentobarbital anesthesia (80 mg/kg i.p. for 45 min) and PC was low all during the day. The PC increase following formalin administration to morphine dependent rats was higher than in naive rats. 0.1 mg/kg s.c. of dexamethasone completely inhibited PC increase after formalin in naive rats, but the dexamethasone inhibition in morphine dependent rats was not complete even with a larger dose, i.e. 10 mg/kg s.c. Diurnal variations and other differences were not detected in the adrenal response to ACTH (in vivo, in vitro) between naive and morphine dpendent rats. These results suggest that HPA function takes part in the development of morphine dependence.


[Thyrotropin-releasing hormone (TRH): Enhancement of dopamine dependent circling behavior and its own circling-inducing effect in unilateral striatal lesioned animals (author's transl)]. Enhancement by TRH of the dopamine(DA) agonist-induced circling behavior and effect of TRH itself on circling behavior were investigated. TRH(2.5--20 mg/kg, i.p.) remarkably enhanced the circling behavior induced by apomorphine or-L-DOPA in the mice lesioned unilaterally in the caudate nucleus by injection of 6-hydroxydopamine (60OHDA) or by tissue aspiration with subsequent reserpinization. TRH also enhanced the apomorphine-induced stereotypy in reserpinized normal mice. The above TRH-enhancing action of the circling behavior was potentiated, suppressed or unaffected by alpha-methyl-para-tyrosine (alpha-MT) or GABA-ergic drugs. In the 6-OHDA lesioned mice treated with TRH, the cyclic AMP formation by DA or apomorphine was clearly enhanced in the triatal slices taken from the lesioned side but not from the intact side. In the rats lesioned unilaterally in the nigrostriatal DA pathway by 6-OHDA, high doses of TRH injected i.p. (100 mg/kg) or into the non-lesioned caudate nucleus (50 micrograms) produced circling toward the lesioned side, which was suppressed by haloperidol or alpha-MT. TRH(10(-5)--10(-3)M) increased the 14C-DA release from the rat striatal slices in vitro. These results suggest that TRH at low doses facilitates the DA postsynaptic transmission in association with an increase of DA-stimulated cyclic AMP formation in the striatum under supersensitization of the DA receptors, and also at high doses enhances the DA neuronal activity by increasing the DA release from the striatal nerve terminals.


[Treatment of orthostatic disorders of circulatory regulation by means of DET MS retard]. 30 patients suffering from troubles in orthostatic circulatory regulation in the sense of a so-called hypersympathicotonic reaction were treated with DET MS retard for 4 weeks and permanently controlled concerning objective changes (circulation by modified Schellong-test on the tilting table) as well as subjective changes (symptoms). The evaluation of the findings shows the stabilisation of the regulation of blood pressure, an extensive economization of the circulatory behaviour during orthostasis, and a statistically evident improvement of clinical symptoms.


[Carcinoma after-care in an after-care clinic based on experiences at the Sonnenberg Clinic, Bad Soden-Allendorf]. The follow-up cure therapy in the framework of cancer follow-up treatment is of special importance. During the past 7 years 7814 patients have been treated according a special regimen in our clinic, which also includes cytostatic and hormonal therapy. Diagnostic control examinations for all patients are performed. Besides adequate medical treatment psychological and social aspects of the individual patient should be considered, furthermore the course of the disease, the question of again taking up life with their families and their partners and of returning to their former professional work. It could be shown that general discussion groups and cancer selfhelp groups, which are organized by the patients, are fruitful. It seems beneficial for the patients to participate in these activities in a follow-up clinic for a course of 6 weeks and they seem to recuperate better regarding their general health and survival time.


Influence of nutritional factors on the renotrophic action of ACTH in the uninephrectomized rat. In the rat, renal compensatory hypertrophy (RCH) was apparent 48 h after uninephrectomy; it was significantly enhanced by long-acting beta1-24-corticotrophin (ACTH) when the animals had free access to food and a NaCl solution (9 g/l). In rats starved after uninephrectomy but drinking the NaCl solution freely, RCH was suppressed: the weights of the body, heart, liver, and solitary kidney were reduced. In similarly starved rats treated with ACTH, the weights of the heart and the solitary kidney were normal. RCH was also impaired in rats fed only a glucose solution (30 g/dl) after uninephrectomy, but it is restored by ACTH, which significantly increases the weight of the remaining kidney. This renotrophic action of ACTH may be related to hyperglycemia and, perhaps, elevated urinary K excretion, which occur in hyper-adrenocorticism and increase the work load of the nephron.


Inhibitory control of the pituitary LH secretion by LH-RH in male rats. Luteinizing hormone-releasing hormone (LH-RH) was administered to prepubertal male rats (intact, castrate or castrate-adrenalectomized, 60 g body weight) for 28 days (1 microgram LH-RH/day, s.c.), at a 10-fold physiological dose, as compared to the minimal FSH-releasing dose of 100 ng/rat s.c. In intact rats, serum LH and weight of androgen-dependent organs (vented prostate, seminal vesicles) were reduced after 14 days of treatment. In castrate rats, the postcastration rise in serum LH was abolished by treatment. Pituitary LH content, FSH secretion and prolactin secretion were not suppressed. Hypothalamic LH-RH was increased at 14 and 21 days. In castrate adrenalectomized male rats, LH secretion was also suppressed by 1 microgram LH-RH s.c. x 28 days. The hypothalamic LH-RH content did not increase. The pituitary LH-RH receptor level was not down-regulated after 14 days treatment either in intact or castrate male rats. Pituitary inhibition (LH release) in rats by a supraphysiological dose of LH-RH given for 28 days indicates that the optimal regime for chronic treatment has to be determined by monitoring LH release at regular intervals. Direct pituitary inhibition by LH-RH may explain some of the unexpected antifertility effects observed with high doses of LH-RH.


The use of artificial beta cell in diagnosis and treatment of insulinoma. The glucose-controlled insulin infusion system (GCIIS), the socalled artificial beta-cell, is an important and useful device for detecting and treating hypoglycemic reactions. The dangers of several diagnostic tests such as the tolbutamide or the insulin response test may successfully be avoided. Patients suffering from severe hypoglycemia are kept in normoglycemia by the feedback-controlled dextrose infusion before and during operation.


Artificial beta-cell application in two cases of insulinoma: a different pattern in beta-cell adenoma and carcinoma. With the aid of an artificial beta-cell (Biostator, Miles Laboratories Inc.), a different metabolic and biological pattern of behaviour was observed in benign versus malignant insulinoma. In the patient with beta-cell adenoma but not in the one with carcinoma, plasma insulin concentrations decreased promptly and markedly, and blood glucose increased during diazoxide and somatostatin infusion. Moreover, only in the adenoma patient was glucose need characterized by a circadian rhythm with the maximum values during daytime. This behavior could reflect the degree of tumor beta-cell differentiation. The controlled glucose and insulin infusion was of great help during and after surgical treatment.


Inhibition of murine sarcoma virus induced transformation in an adenovirus -- NRK system. Cell transformation induced by murine sarcoma virus (MSV-M) is significantly inhibited (80--90%) in a clonal line of normal rat kidney (NRK) cells when they are infected with rat cell passaged adenovirus 12 (R-Ad12). No inhibition is seen when R-Ad12 is added simultaneously with, or 1 1/2 or 24 hr after, MSV-M infection, suggesting that inhibition occurs most probably intracellularly. There is also a direct correlation between the extent of focus formation and the concentration of R-Ad12 used. Concentrations of R-Ad12 used to inhibit cell transformation do not affect cell growth. Significant inhibition in foci (90--100%) is also evident in the first two subcultures of R-Ad12 infected NRK cells. The mechanism of the inhibition is not yet known.


Isolation of alveolar type II cells by centrifugal elutriation. Centrifugal elutriation (counterflow centrifugation) was used to develop a reproducible method for obtaining a nearly pure population of isolated alveolar type II cells. Lung was dissociated into individual cells with recrystallized trypsin, and the type II cells were partially purified by centrifugation on a discontinuous density gradient. The alveolar type II cells were finally purified by centrifugal elutriation. Cells were collected from the elutriator rotor by stepwise increases in flow rates. Cells obtained at flow rates of 7 and 14 ml per min were lymphocytes, other small cells, a few type II cells and cell debris; cells collected at flow rates of 18 and 22 ml per min were mainly type II cells; and cells collected at flow rates of 28, 34 and 43 ml per min were macrophages, some type II cells, other lung cells and cell aggregates. At flow rates of 18 and 22 ml per min, 1.9 +/- 1.0 x 10(6) cells per rat lung (mean +/- S.D., n=30) were recovered of which 86 +/- 6% were type II cells. At these flow rates, 94% of the cells excluded the vital dye erythrosin B from their cytoplasm. They consumed oxygen at a rate of 101 +/- 21 nmol per hr . 10(6) cells (mean +/- S.D., n=4), and their oxygen consumption increased only 10% after 10 mM sodium succinate was added. The cells incorporated [14C]leucine into protein and lipid for 4 hr. Electron micrographs of the cells collected at flow rates of 18 and 22 ml per min show a high percentage of morphologically intact alveolar type II cells. We conclude that centrifugral elutriation is a reproducible method for obtaining nearly pure, metabolically active alveolar type II cells.


Inhibition of cellular transition from G1-resting to G1-prereplicative phase by aminonucleoside of puromycin. Human embryonic lung fibroblasts (IMR-90 and WI-38) were arrested in the G1 phase of the cell cycle by serum deprivation and high population density. Within 1 hr after the addition of medium containing fresh serum, these cells showed an increase in rRNA synthesis. The inclusion of 100 micrograms per ml aminonucleoside of puromycin (AMS) in the fresh medium eliminated the serum stimulation of rRNA synthesis and prevented the cells from making the G1-resting phase to G1-prereplicative phase transition. AMS also prevented the synthesis of HnRNA normally found within 10 hr after serum stimulation. Serum-stimulated RNA synthesis in starved, SV-40 transformed fibroblasts (WI-38-VA-13 cells) was inhibited, but not completely prevented, by AMS indicating that transformed cells may produce specific RNA's that are not AMS-sensitive and that may be responsible for the failure of transformed cells to be arrested in G1.


High-density lipoprotein cholesterol concentration and other serum lipids in an isolated island community free of coronary heart disease. Fasting serum high-density lipoprotein (HDL) cholesterol, other lipid concentrations and related characteristics have been measured in 87% of 261 residents aged over 5 years on the small isolated island of Salt Cay, Turks and Caicos Isles. Only one subject, a hypertensive woman, had a major electrocardiographic Q wave abnormality, and none of the islanders had clinical coronary heart disease. These findings, together with local clinical experience indicated a low incidence of coronary heart disease in this community. Adult mean concentrations of HDL cholesterol (1.6 mmol/l) and serum total triglyceride (0.81 mmol/l) were relatively high and low respectively compared with those of men and women in North American communities. Unlike findings in North America, there was no significant sex difference in HDL cholesterol concentration apparent in adulthood and this was not explained by sex differences in alcohol consumption (positively correlated with HDL cholesterol in both sexes) or adiposity (negatively correlated with HDL cholesterol in men only). Residents who were recovering from an epidemic of mild upper respiratory infection had on average a 9% reduction in HDL cholesterol concentration compared with the remainder of the community. The lipoprotein lipid pattern in these people is consistent with a low cardiovascular risk status, and might account for the apparent absence of coronary heart disease on the island.


Transition of nervous activity from signals to information processing. The author starts from the fact that although the signals are the information bearers, they cannot be identified with the information they are carrying. Therefore, in order to achieve control as adequate as possible of human behaviour, the nervous system has to discover the information brought by different signals. The activity of information discovery is realized by means of superisation process, that is by means of transition from many lower order signals to a superior order signal. By means of the superisation process a gradual recognition of the respective signals emitting sources is realised. This recognition is made with the help of certain logical circuits representing the models of different sources written in the structure of the neuronal network. In this manner the nervous system passes step by step from control by means of signals to control by means of information by which the superior structures thereby exert much more subtle control, and supervise the inferior structures working by signals. But, because this control cannot remove all the errors of the inferior structures, different psychical and psychosomatic illnesses can occur.


Effects of interstimulus interval on latency of the brainstem auditory evoked potential. Peak latency of the inferior collicular response (wave V) of the brainstem auditory evoked potential (BAEP) increases with decreasing interstimulus interval (ISI). The present research investigated relative peak shifts of major components of the BAEP in an attempt to identify the source of the observed shift in wave V. Peripheral as well as central transmission times (CTT) were computed for 9 young adults between an ISI of 100 msec and an ISI of 10 msec. The results indicate that shifts in wave V latency with changing ISI are due to a combination of central and peripheral processes. The magnitude of the shift observed for wave V was approximately 0.50 msec. This increase in latency was accounted for by an average 0.29 msec increase in peripheral transmission time, a 0.14 msec increase in CTT between waves I and III, and a 0.07 msec increase in CTT between waves III and V. The results suggest a dual process underlying latency changes in wave V of the BAEP as a function of decreasing ISI. These are peripheral adaptation or fatigue and central processes that may include short-term habituation.


Effect of diethylstilbestrol on replication and transformation by human herpesviruses. Human embryo fibroblasts treated with diethylstilbestrol (DES) gave rise to slightly increased plaque production by cytomegalovirus (CMV): plaques were also slightly larger in treated cells when compared to those in untreated cultures. There was no significant enhancement of plaque production by herpes simplex virus (HSV) types 1 and 2 in the human cells or in primary rabbit kidney cells treated with DES. Growth analyses of HSV and CMV in DES-treated cells failed to reveal enhancement of virus production when compared to untreated cells. The frequency of biochemical transformation of mouse cells deficient in thymidine kinase (TK) to the TK+ phenotype by HSV was markedly enhanced when cells were pretreated with DES. TK+-transformed colonies arising from DES-treated cells were also larger than those derived from untreated cultures. These reveal that DES increases susceptibility of human cell lines to CMV infection and enhances efficiency of biochemical transformation of mouse cells by HSV.


Cervical cancer cell lines containing herpesvirus markers. Five epithelioid cell lines were established from invasive cervical carcinoma tissue and were examined for herpes simplex virus (HSV) antigens or virus-induced changes. Immunofluorescence studies with antiserum to an HSV type 2 nonstructural polypeptide detected perinuclear antigens. In these cell lines, concentrations of a membrane fucopeptide that is regularly present in normal cells were reduced to levels similar to those previously observed in virus-transformed cells. One of the cell lines inoculated into athymic mice produced tumors.


Effect of antipain on radiation induction of endogenous type-C virus from mouse cells in vitro. Xenotropic murine type-C virus was induced after X-ray or UV light irradiation of K-Balb cells. The induction was markedly suppressed by postirradiation treatment of the cells with antipain, a protease inhibitor of microbial origin. Virus expression was unaffected when the cells were treated with halogenated pyrimidines.


Chemodectomas: a review of 17 cases.

Chemodectomas: a review of 17 cases. Seventeen cases of chemodectomas seen in the University College Hospital, Ibadan, are presented. Eight of them (48%) are carotid body tumors, while six (36%) are glomus jugulare. The rest are subglottic, abdominal and multicentric. The sex distribution for the two major types is equal for males and females; but they all seem to appear in younger age groups. Operative mortality has been exclusive to carotid body tumors (50% for this group), while bleeding is the leading postoperative complication in all the cases. The embryology, pathophysiology and clinical presentation of chemodectomas are discussed and it is suggested that, in the management of this condition, the advantages of operation should be weighed against morbidity and mortality risks.


Foot-and-mouth disease virus immunogenic capsid protein VPT: N-terminal sequences and immunogenic peptides obtained by CNBr and tryptic cleavages. The immunogenic capsid protein (VPT), circa 30 kiladaltons (kd), of foot-and-mouth disease virus was examined for (i) its ability to induce neutralizing antibody in guinea pigs after chemical modifications and CNBr or tryptic cleavages and (ii) N-terminal amino sequence homology across three virus types. The immunogenicity of VPT was inactivated by glutaraldehyde treatment, carboxymethylation and maleylation or citraconylation. However, de-citraconylation restored part of the lost activity. Cleavage of type A12 VPT with CNBr produced an immunogenic peptide of circa 13 kd. A slightly larger (ca. 16 kd) immunogenic doublet, VPTab, was obtained by tyrptic cleavage of VPT in the virion. Sequence homologies of circa 85% were found between the first 26 amino acids at the N-terminus of VP chains from virus types A12 strain 119 (A12), C3 Resende (C3R) and O1 Brugge (O1B).


Comparative study of herpes group virus-induced DNA polymerases. A comparative biochemical study of virus-induced DNA polymerases was made among the herpes group viruses: namely, herpes simplex virus (HSV) type 1 and type 2, human cytomegalovirus (HCMV) and varicella-zoster virus (VZV). Although these virus-induced enzymes shared some biochemical properties, they differed in several important aspects. All these virus-induced DNA polymerases could efficiently use poly(dC) . oligo(dG)12--18 and poly(dA) . oligo(dT)12--18 as template-primers. However, in phosphocellulose chromatography, HSV-1- and HSV-2-induced enzymes were eluted at the low concentration of 0.18--0.20 M NaCl and the counterparts of HCMV and VZV were eluted at 0.30--0.32 M. The former two enzymes were more sensitive to lower concentrations of phosphonoacetate and ethyl phosphonoacetate than the latter two enzymes. Moreover, the activity of HSV-1- and HSV-2-specified DNA polymerases was 5 times greater in the presence of 60 mM ammonium sulfate if poly(dA) . oligo(dT)12--18 was used as template-primer, while HCMV- and VZV-induced enzyme activities were only about twice as great under the same conditions. Futhermore, DNase activity was conspicuous in both HSV-1- and HSV-2-infected WI-38 cells, but was not detectable in HCMV- and VZV-infected cells. After storage for 1 year at 4 degrees, the HSV-1-induced DNA polymerase was the most thermostable of the four viral enzymes.


Cytomegalovirus isolations from cell cultures of human adenocarcinomas of the colon. Cytomegalovirus was isolated from cell cultures derived from 3 of 16 surgical specimens of adenocarcinomas of the colon. Virus identification was accomplished through electron microscopical, cytochemical, and immunofluorescent procedures.


Subculture requirement for rescue of SV40 from 'nonrescuable' cell lines. Virus could be rescued by cell fusion techniques from the H50, BRKSV (Bam I linear), 14B and 14B (1--4) cell lines, which contain the entire SV40 genome but have previously yielded negative results by this method. Virus could not be rescued from the T22 cell line, which was transformed by defective virus. The T-antigen-negative flat revertant cell line, 14B (1--4), converted to T-antigen-positive prior to rescue. Subculture and extensive monitoring for production of infectious virus appeared to be prerequisite for detecting virus rescue.


[Epidermolysis bullosa dystrophica inversa, a review and case report]. An additional case of epidermolysis bullosa dystrophica inversa is reported. A review of the literature shows that until now reports about probably 15 patients from seven families were published, although the first 13 patients were not investigated by electron microscope [3]. By blistering beneath the basal lamina similar to the Hallopeau-Siemens type these patients develop bullae predominantly at the trunk and the inverse sites of the axillar and genitoanal regions continuously or at close intervals starting shortly after birth. The mucous membranes of the mouth, the esophagus, and the cornea and conjunctiva of the eye are very often involved. By bacterial infection of the skin lesions the patients develop hpochrome anemia. The web-like structures in the esophagus following blistering can be successfully treated by careful dilatation because of the low tendency to scar formation. An autosomal recessive mode of inheritance seems to be most likely.


Adrenocortical function in rats chronically exposed to high altitude. In rats exposed to a simulated altitude of 5,486 m for 3 mo, pituitary and adrenal glands hypertrophied and plasma levels of corticosterone increased more than threefold over sea-level controls. The in vitro rates of corticosterone production by the quartered adrenal gland were significantly enhanced, but the responsiveness of the adrenal gland to ACTH remained normal.


Effects of hypophysectomy and dexamethasone on rat adrenal response to microwaves. Circulating corticosterone levels were measured to compare the adrenocortical response to acute microwave exposure of normal, hypophysectomized, or sham-hypophysectomized rats. Plasma corticosterone levels in acutely hypophysectomized rats exposed to 60 mW/cm2 for 60 min were below control levels, indicating that the microwave-induced corticosterone response observed in normal, intact rats is dependent on ACTH secretion by the pituitary. In other groups of rats pretreated with dexamethasone before being exposed to microwaves for 60 min, the corticosterone response to a 50-mW/cm2 exposure was completely suppressed by doses equal to or greater than 3.2 micrograms dexamethasone/100 g body weight. However, the corticosterone response to a 70-mW/cm2 exposure was only partially suppressed by prior administration of 3.2 or 5.6 micrograms dexamethasone/100 g BW. The evidence obtained in these experiments, in conjunction with the results of other experiments previously reported, is consistent with the hypothesis that the stimulation of the adrenal axis in the microwave-exposed rat is a systemic, integrative process due to a general hyperthermia.


Ventilatory responses to CO2 and lung inflation in tonic versus phasic REM sleep. Ventilatory responses to CO2 and to lung inflation were compared in four dogs during tonic and phasic segments of rapid-eye-movement (REM) sleep. Phasic REM sleep (P-REM) was identified by the presence of bursts of rapid eye movements, visible muscle twitchings, and frequent phasic discharges in the nuchal electromyogram. These features were absent during tonic REM sleep (T-REM). During P-REM the response of minute volume of ventilation (VI) to progressive hypercapnia (0.58 +/- 0.19 (l/min)/Torr, mean +/- SE) was significantly less than in slow-wave sleep (SWS) (1.40 +/- 0.14; P less than 0.05). In contrast, during T-REM the response (1.48 +/- 0.19) was similar to that in SWS. Similarly, during P-REM the duration of apnea (5.9 +/- 1.5 s) elicited by sustained inflation of the lungs with 1.0 liter of air, was significantly shorter than in SWS (25.8 +/- 0.8); in contrast, during T-REM the duration of apnea (17.8 +/- 3.6) was similar to that in SWS. The results indicate that previously described decreases in VI responses to CO2 and apneic responses to lung inflation during P-REM, compared to SWS, are related to the phasic phenomena of REM sleep, rather than to the REM sleep state per se.


Development, preparation and safety testing of a Clostridium welchii type C toxoid. I: preliminary observations in man in Papua New Guinea. The reactogenicity and immunogenicity of various strengths of plain and adsorbed Cl. welchii type C toxoid have been evaluated in laboratory tests and in man in Papua New Guinea. The greater antigenicity and acceptable clinical reactivity of a vaccine containing 50 total combining power units per 0.5 mm of adsorbed toxoid resulted in its selection for further field studies.


Development, preparation and safety testing of a Clostridium welchii type C toxoid. II. Laboratory evaluation of potency. The results obtained with four laboratory tests on four candidate formulations of Clostridium welchii type C vaccine for use in man have been compared with clinical responses to the same vaccines. Quantal response assays in mice appeared to reflect the ranking of the four vaccines in human subjects better than did the guinea pig tests. They also enabled the potency of the vaccine preparations to be related to an existing International Reference Preparation. Mouse assays in which the animals received two spaced doses of vaccine prior to challenge yielded marginally more satisfactory results in terms of precision and reflection of human responses than did assays involving a single dose of vaccine.


The effects of EGTA and trypsin on the serum requirements for cell attachment to collagens. Cells growing on plastic or glass surfaces in vitro may be brought into suspension by proteases (e.g. trypsin) or chelating agents (e.g. EGTA). Trypsin and EGTA remove different quantities and types of molecules from cell surfaces. Previous studies have revealed that when confluent cultures of either BHK or PyBHK cells are brought into suspension by exposure to trypsin, foetal calf serum (or fibronectin) is required for cell attachment to films of denature type I collagen, but not to 3-dimensional gels of native collagen fibres. In this communication the serum requirements for the attachment of BHK and PyBHK cells to collagen substrata have been examined as a function of (a) the method used to prepare the cell suspension (EGTA or trypsin), and (b) cell density. Data are presented consistent with the view that cell surface-associated fibronectin is able to mediate cell attachment directly to films of denatured collagen.


Cholinergic inhibition of catecholamine-stimulable cyclic AMP accumulation in murine atria. Carbachol antagonizes isoproterenol-stimulable cyclic AMP accumulation in mouse atria by direct activation of cardiac muscarinic receptors. Inhibition by carbachol occurs rapidly and is completely reversed when the drug is removed. Neither nitroprusside nor 8-bromo-cyclic GMP mimics the actions of carbachol and low concentrations of carbachol block cyclic AMP accumulation without increasing the intracellular cyclic GMP content. Carbachol does not block cyclic AMP accumulation by activating phosphodiesterase since it is fully effective in the face of marked phosphodiesterase inhibition, nor does it appear to inhibit the catalytic activity of adenylate cyclase since it does not decrease either basal or cholera toxin-stimulated cyclic AMP accumulation. The interaction between carbachol and isoproterenol is not competitive, since cholinergic inhibition cannot be surmounted by increasing concentrations of isoproterenol. The site of muscarinic action therefore appears to involve the mechanisms coupling the hormone-receptor complex to adenylate cyclase. This site is distinct from that of cholera toxin action since there is no antagonism between the effects of cholera toxin and carbachol on cyclic AMP metabolism in the atrium.


Characterization of immunofluorescent cyclic GMP-positive fibres in the central nervous system. The cyclic GMP immunofluorescent fibres in the rat central nervous system have been characterized as processes of fibrous astrocytes, on the basis of distribution and similarity to the localization of glial fibrillary acidic protein. The neuroglial localization of the nucleotide is discussed together with the surprising observation that these cyclic GMP positive fibres are absent from the central nervous system of the adult mouse.


Prostacyclin increases cAMP in coronary arteries. This study was designed to determine whether prostacyclin (PGI2)-induced relaxation in circular strips of coronary arteries might be mediated by cAMP. Partially depolarized circular strips of bovine coronary arteries were used and PGI2-induced changes in length were compared with tissue concentrations of cAMP and cGMP, measured by RIA. It was found that PGI2 produced significant and concentration dependent increases in cAMP levels which were closely associated with the relaxant effects produced by the same concentrations (0.3 - 26.7 muM). Cyclic GMP was not changed by these concentrations. The relaxant effects of PGI2 were not antagonized by propranolol. There was a significant linear correlation between log increases in cAMP and percent relaxation produced by PGI2 which was almost identical with similar correlations obtained with either isoprenaline or adenosine, indicating that the relaxant effects of PGI2 are in analogy to those of isoprenaline and adenosine likely to be mediated by cAMP.


Assay for vitamin D2 and vitamin D3 in plasma of dairy cows: changes after massive dosing of vitamin D3. A sensitive, precise assay for vitamin D in plasma is described. Three to five milliliters of plasma were extracted with methanol:methylene chloride (2:1). The lipid extract was chromatographed on Sephadex LH-20 and then on lipidex-5000 columns. After high pressure liquid chromatography with a reverse phase chromatographic system, vitamin D2 and vitamin D3 were quantitated by ultraviolet absorbance. We used this assay system for monitoring daily changes of vitamin D3 in plasma of two Jersey cows after four intramuscular doses (15 x 10(6) IU) of vitamin D3 administered at weekly intervals. Basal vitamin D in plasma was 3.2 +/- .99 ng/ml with a range of 1.7 to 4.9 ng/ml. Vitamin D3 in plasma remained relatively low (10 to 45 ng/ml) the week after the first vitamin D3 injection. Vitamin D3 was high (130 to 234 ng/ml) after the second, third, and fourth injections. Vitamin D3 decreased steadily to 88 ng/ml by 38 days after the fourth vitamin D3 injection. Phosphorus in plasma increased sharply to a plateau at 9.5 mg/100 ml during the week after the second vitamin D3 injection and returned to normal (4.5 mg/100 ml) at the end of the experiment. Calcium, however, gradually increased to 14.0 mg/100 ml 20 days after the fourth vitamin D3 injection. Both animals remained hypercalcemic (calcium 11.5 mg/100 ml) during the experiment.


Adrenal response to adrenocorticotropin in Holstein heifers exposed to a cool environment. Tne Holstein heifers acclimated to either -2.7 or 19.1 C were used to determine glucocorticoid concentrations in plasma following injection of adrenocorticotropin (200 IU). Environmental treatments did not change glucocorticoids following adrenococorticotropin. Adrenocorticotropin injection elevated mean glucocorticoids to 41.6 ng/ml plasma at -2.7 C and 42.8 ng/ml at 19.1 C. Adrenal response to adrenocorticotropin in Holstein heifers was not altered by low environmental temperature.


The spread of rhinoviruses in the Soviet Union (USSR) and in Czechoslovakia (CSSR) and the role of their pathology. Virological, serological and immunofluorescence studies revealed circulation of rhinoviruses of the strains 1A, 1B, 2, 3, 7, 9, 10, 12, 13, 14, 16-23, 27, 29-33, 42, 48, 53, 56, 60 and 69 on the territory of Czechoslovakia and the Soviet Union. According to virological results, type 48 predominated and was followed in frequency of occurrence by types 27, 14 and 16 in the USSR and 30, 1A and 31 in the CSSR. RV infection in adults with ARD diseases was the aetiology in 28.5% of cases. Among the causative agents of acute rhinitis, rhinosinusitis and otitis, rhinovirus infection makes up a large proportion. Interrelation has been found between rhinovirus infection and chronic inflammation of the mucosa of the nose, the accessory sinuses and the ears. Prolonged course of rhinovirus infection and isolation of the virus in the late stages of disease (1-4 months after its onset) support the presumption that rhinoviruses promote chronicity of inflammation. RV was isolated from patients with chronic pneumonia from the lower respiratory passages (bronchial secretion). Rhinoviraemia was found in a child who died of ARD.


Immunological surveys of antibodies against B. pertussis and B. parapertussis in some African and Asian countries. Immunological surveys in African and Asian countries showed a different degree of herd immunity in the respective countries and the circulation of various types of B. pertussis. Antibodies against B. parapertussis, the second aertiological agent of whooping cough, were found in all countries in all age groups. Before planning any vaccination program, attention should be paid to the vaccine concerning the content of all types of B. pertussis, as was shown in the results of the testing of different vaccines used in Mongolia and Algeria. It is also possible to estimate the age limit for vaccination and thus economic use of the vaccine. The contemporary state of reporting whooping cough cases is very unsatisfactory.


Evidence of specific nuclear binding sites for T3 in the mouse cultured fibroblast. Mouse L 929 cultured fibroblasts revealed saturable and high affinity nuclear receptors for triiodothyronine. 125I-T3 bound rapidly to intact cells, with a steady state achieved between 1-2 h at 37 C. By Scatchard estimation, the maximal binding capacity averaged 18 fentomoles of T3 bound per 100 microgram of DNA, which approximated to 2,000 sites per cell nucleus; the apparent equilibrium association constant Ka averaged 4.90 x 109 LM-1. The relative affinity of T3 analogs tested correlated with their respective thyromimetic potency. Limited capacity, high affinity and specificity of T3 nuclear receptors in these fibroblasts were found to have properties similar to those previously observed in other cell lines or tissues. Such cultured fibroblasts may provide a useful experimental model to investigate the intracellular biological effects of thyroid hormones.


Induction of androgen-dependent protease and serous-like granules by tri-iodothyronine in the submandibular gland of mice with testicular feminization. Esteroprotease, an androgen-dependent enzyme of the mouse submandibular gland, was increased by injection of tri-iodothyronine (T3) in mice with testicular feminization (Tfm) which are genetically deficient in androgen receptors. Histochemical and electron microscopic studies also demonstrated increases of RNA and serous-like granules in cells of the convoluted tubules of the gland. These findings suggest that the esteroprotease gene in Tfm mice is normal and that T3 can induced both esteroprotease and serous-like granules independently of androgen.


Properties of rat adrenal zona reticularis cells: production and stimulation of certain steroids. The outputs of corticosterone, deoxycorticosterone and androstenedione from dispersed, purified rat adrenal zona reticularis and zona fasciculata cells have been measured by radioimmunoassay. Preferential production of deoxycorticosterone by zona reticularis cells was demonstrated by their higher basal deoxycorticosterone : corticosterone ratio when compared with zona fasciculata cells. Adrencorticotrophin (ACTH) stimulated corticosterone output by all cell pools prepared by unit gravity (1 g) sedimentation, zona fasciculata cells being stimulated 130-fold compared with 20-fold for the zona reticularis cells in relation to their basal corticosterone output. In every cell pool, ACTH stimulated the output corticosterone more than it stimulated the output of deoxycorticosterone. In parallel cell preparations, it was shown that ACTH increased the conversion of tracer amounts of radioactive deoxycorticosterone to corticosterone and decreased the conversion of radioactive corticosterone to 11-dehydrocorticosterone. Adrenocorticotrophin did not increase the conversion of radioactive deoxycorticosterone to total 11-oxygenated steroids (corticosterone + 11-dehydrocorticosterone). It is unlikely therefore that ACTH stimulates 11 beta-hydroxylation. Data indicate that the ratio of deoxycorticosterone to total 11-oxygenated steroids (corticosterone + 11-dehydrocorticosterone) is characteristic for each cell type, and that this ratio will be relatively independent of ACTH stimulation or the amount of pregnenolone substrate available. Basal androstenedione outputs were similar for both types of cell, and ACTH stimulation was very small, being slightly greater for zona fasciculata than for zona reticularis cells. The contribution of the zona reticularis cells to the basal output of any steroid by the cells of the inner two zones of the adrenal cortex of the rat was relatively small (20% for deoxycorticosterone and 10% for corticosterone) and was even less after stimulation by ACTH. Unless a specific stimulus can be found, therefore, a significant role for the zona reticularis cannot yet be established.


Occupations of fathers of patients with Wilms's tumour. A case-control study of 149 Connecticut-born children with Wilms's tumour reported to the Connecticut Tumor Registry during the period 1935--1973 and of 149 matched controls was undertaken in order to explore the possibility that children with Wilms's tumour may have been exposed perinatally to carcinogenic agents. The occupation of the father at the time of the child's birth was investigated and used as an indicator of potential sources of carcinogens to which infants in the study may have been exposed. An association was found between paternal occupations related to lead in the group developing Wilms's tumour compared with the controls.


Responses of the early chick embryo to external cAMP sources. Early chick embryos were stimulated with local sources of cAMP. Three major effects were observed: bending of the embryonic axis, attraction of cells on the ventral surface of the embryo, and disruption of the blastodisc. Each had a characteristic concentration dependence. These results are compared with those from studies of cells disaggregated from similar embryos.


Neuromuscular transmission in Peripatus. The electrical and mechanical responses of body, leg and jaw muscle of Peripatus to electrical excitation of their motor-nerves were examined. A small twitch was obtained from each muscle, whose strength increased stepwise with increasing stimulus strength. In jaw and body muscles as many as ten increments in height were obtained with increasing stimulus strength. Only a single twitch height was obtained from the claw retractor muscle. Tetanus:twitch ratio under supraminal stimulation was less than 2:1 for jaw muscles, about 50:1 for the claw retractor and about 6:1 for muscles moving the legs. The jaw-muscle twitch duration was 0.6 s, that for the leg muscles 1.2 s and for body muscles about 3.0 s. Large miniature junctional potentials were frequently recorded regardless of electrode location. Responses to neural stimulation consing stimulus strength, generally with three steps. With repetitive stimulation, facilitation of the second and third junctional potentials occurred, plus summation. A few fibres gave spikes to a single shock: most gave a few spikes sporadically, during repetitive stimulation only. No abrupt tension increments occurred in whole muscles when individual fibres spiked. We saw no evidence for peripheral inhibitory axons. The excitation of Peripatus muscle is by local graded junctional potentials at distributed nerve-on-muscle fibre synapses, together with action potentials. The latter are initiated only by larger junctional potentials compounded of multiple smaller ones summated and/or facilitated. The details of neuromuscular physiology are not compatible with the phylogenetic status commonly proposed for Peripatus.


Acute and chronic hypoxia in rats. II. Effect on oxidative phosphorylation and in vitro protein synthesis in liver mitochondria and its subpopulations. The objective of this investigation was to examine liver mitochondrial functions in rats exposed to 0.4 atm for 0, 5 and 27 days, Liver homogenates were fractionated by rate-zonal centrifugation utilizing iso-osmotic Ficoll-sucrose gradients; this eliminates loss of large and small mitochondria and makes possible the separation of mitochondria into subpopulations according to sedimentation coefficient. After pooling all mitochondrial fractions for obtaining composite determinations of the entire population, large diminutions in states 3 and 4 respiration (succinate as substrate) were obtained in day-5 and day-27 rats but no changes were evident with regard to ADP:O ratios, respiratory control indices or the capacity for in vitro protein synthesis. By examination of subpopulations of mitochondria, it was found that mitochondria are heterogeneous with regard to ADP:O ratios, respiratory control indices, states 3 and 4 respiration and the capacity for in vitro protein synthesis. The heterogeneity for each of these parameters was altered in day-5 and day-27 animals. Although states 3 and 4 respiration were depressed throughout the entire mitochondrial population for day-5 and day-27 rats, a subpopulation of mitochondria from day-27 rats showed respiratory control indices and ADP:O ratios which were higher than any subpopulation of mitochondria of either day-5 or day-0 animals.


The nonlinear pathway of Y ganglion cells in the cat retina. Retinal ganglion cells of the Y type in the cat retina produce two different types of response: linear and nonlinear. The nonlinear responses are generated by a separate and independent nonlinear pathway. The functional connectivity in this pathway is analyzed here by comparing the observed second-order frequency responses of Y cells with predictions of a "sandwich model" in which a static nonlinear stage is sandwiched between two linear filters. The model agrees well with the qualitative and quantitative features of the second-order responses. The prefilter in the model may well be the bipolar cells and the nonlinearity and postfilter in the model are probably associated with amacrine cells.


Labeling of hepatitis B virus surface antigen (HBsAg) synthesized in a HBsAg-producing hepatoma cell line. Hepatitis B virus surface antigen (HBsAg) could be studied until recently only by isolating it from the blood of carriers, thus making incorporation of radioactive precursors into this protein(s) impossible. The isolation of a cell line producing HBsAg [Alexander et al, 1978] has eliminated this obstacle. The cell line was therefore used for labeling HBsAg either with 35S-methionine or with 35S-cystine. HBsAg was purified by pelleting the component and by isopycnic centrifugation in CsCl gradients. HBsAg-positive fractions (as determined by solid-phase radioimmunoassay) were isolated from the gradients and analyzed in sodium dodecyl sulfate-containing polyacrylamide gels. It was found that although HBsAg contains substantial amounts of 35S-cystine, very little 35S-methionine was incorporated into this protein. In contrast, both labels were found in other structures having a buoyant density of about 1.3 gm/cm3 in CsCl. It was concluded that HBsAg is very low in methionine, and therefore this amino acid should not be used for labeling HBsAg in cells or in a cell-free system. Analysis of 35S-cystine-labeled HBsAg-positive material (buoyant density about 1.2 gm/cm3 in CsCl) revealed five proteins with molecular weights in the range of 48,000-82,000.


The mode of action alginic acid compound in the reduction of gastroesophageal reflux. This study was designed to evaluate quantitatively the mode of action of alginic acid compound (AAC) in the treatment of patients with symptomatic gastroesophageal reflux. Gastroesophageal scintigraphy using an orall administered Tc-99m sulfur colloid solution was used to demonstrate that AAC decreased significantly the gastroesophageal reflux index from (9.9 +/- 1.3) % to (6.5 +/- 0.8) % (p less than 0.05). No alteration of lower esophageal sphincter pressure was observed. After ACC was suitably labeled with Sr-87m, a dual-nuclide scintigraphic technique was used to show that most (greater than 75%) of the AAC was located in the upper half of the stomach in both normal subjects and patients with gastroesophageal reflux. In those subjects in whom reflux did occur after treatment with AAC, the Sr-87m-AAC refluxed into the esophagus preferentially compared with the liquid containing Tc-99m sulfur colloid. These findings suggest that AAC dimishes gastroesophageal reflux by means of its foaming, floating, and viscous properties.


Quantitative lymphoscintigraphy I: Basic concepts for optimal uptake of radiocolloids in the parasternal lymph nodes of rabbits. The activity-size distribution of radiocolloids has been studied using gel-chromatography scanning (GCS) of columns filled with Sepharose 4B gel. Rabbits were injected subcutaneously with the colloid of interest, laid supine beneath a gamma camera, and imaged every 15 sec for 2 to 4 hr. From the stored data, the uptakes in the parasternal lymph nodes were analyzed in terms of two-compartment model, and the rate constants measured. The substances tested were Au- 198 colloid, Tc-99m antimony sulfide colloid, Tc-99m tin colloid, Tc-99m phytate, and Tc-99m sulfur colloid. It was shown that the optimal particle size for the colloid in the range 1-10 nm. The large and most rapid uptake was found for Au- 198 colloid, with a particle size of 5 nm, which appeared as a single peak in the GCS spectrum. The percentage uptake after 2 hr for Au- 198 colloid 8%, while it was 5% for antimony-sulfide colloid, which was the best of the Tc-99m-labeled colloids. The GCS spectrum for the antimony product showed a single-peaked size distribution with a somewhat broader range: 5-15 nm. The particles of the other colloids were either too large to pass into the lymphatic system, or too small to be trapped.


Myocardial uptake of technetium-99m stannous pyrophosphate in experimental viral myopericarditis. Distribution of technetium-99m stannous pyrophosphate was studied in mice with experimentally induced viral myopericarditis. Myocardial and bone uptakes of Tc-PPi were compared in 55 mice inoculated with coxsackievirus B3 (Nancy strain). The myocardium-to-bone uptake ratio in 33 mice with myopericarditis was increased to a greater extent than that seen in 22 mice without myopericarditis (p less than 0.001). In the severely involved heart, the uptake per gram exceeded that in the bone. Myocardial uptake in myopericarditis can be visualized on a whole-body image using a pinhole collimator and a left lateral view. Our experimental studies suggest the potential clinical usefulness of myocardial scintigraphy in viral myopericarditis.


1-aminocyclobutane[11C]carboxylic acid, a potential tumor-seeking agent. 1-Aminocyclobutane[14C]carboxylic acid [C-14) ACBC] was incorporated preferentially by several tumor types in rats and hamsters. The agent was cleared rapidly from rat blood, attaining its maximum tissue concentrations within 30 min after i.v. injection. Carrier ACBC had little effect on the tissue distribution of (C-14) ACBC. This agent showed no affinity for a Staphylococcus aureus abscess in rats. The total excretion was low, 3.6% in 2 hr. (C-11) ACBC was synthesized in amounts up to 415 mCi (55% chemical yield) using our modified Bücherer-Strecker technique. Forty minutes were required for the two-step synthesis and chromatographic purification. ACBC was found to be nontoxic in three animal species. The radiation dose from (C-11) ACBC should be minimal. (C-11) ACBC thus appears to have good potential as a tumor-seeking agent, particularly when used with a positron emission computed tomograph.


Potential column chromatography generators for ionic Ga-68. I. Inorganic substrates. Chemical separations for Ga-68 from Ge-68 using adsorption chromatography on inorganic materials are described. The adsorbents used were TiO2, ZrO2, and SiO2. Distribution coefficients for Ge and Ga on these absorbents were determined as a function of reagent concentration and duration of equilibration. The distribution coefficient (w/w) for Ge on SiO2 reached 250 in 6 N HNO3, whereas Ga was no significantly adsorbed. Therefore, Ga-68 can be collected with a mall volume of 6 N HNO3 eluent. By contrast, large volumes of 1 N HNO3 were necessary to collect Ga-68 from ZrO2, since the KD of Ga under these circumstances was about 50. The Ga-68 eluted from TiO2 was chemically contaminated with titanates and would require additional chemical manipulation in order to make it injectable. All the adsorbents could lead to chromatographic systems that would allow acceptable chemical separations. However, the specific requirements for a radionuclide generator, usable in a hospital environment, make the SiO2-based system the most attractive.


Use of blood-pool imaging in evaluation of diffuse activity patterns in technetium-99m pyrophosphate myocardial scintigraphy. It has been suggested that diffuse Tc-99m pyrophosphate precordial activity may be due to persistent blood-pool activity in routine delayed views during myocardial imaging. To answer this question, we reviewed myocardial scintigrams recorded 60--90 min following the injection of 12--15 mCi of Tc-99m pyrophosphate for the presence of diffuse precordial activity, and compared these with early images of the blood pool in 265 patients. Diffuse activity in the delayed images was identified in 48 patients: in 20 with acute myocardial infarction and in 28 with no evidence of it. Comparison of these routine delayed images with early views of the blood pool revealed two types of patterns. In patients with acute infarction, 95% had delayed images that were distinguishable from blood pool either because the activity was smaller than the early blood pool, or by the presence of localized activity superimposed on diffuse activity identical to blood pool. In those without infarction, 93% had activity distribution in routine delayed views matching that in the early blood-pool images. The usefulness of the diffuse TcPPi precordial activity in myocardial infarction is improved when early blood-pool imaging is used to exclude persistence of blood-pool activity as its cause. Moreover, it does not require additional amounts of radioactivity nor complex computer processing, a feature that may be of value in the community hospital using the technique to "rule out" infarction 24--72 hr after onset of suggestive symptoms.


Ovarian imaging by the binding of radioiodinated chorionic gonadotropin: concise communication. Previous studies have demonstrated the presence of specific, high-affinity receptors for human chorionic gonadotropin (HCG) in human and rat ovaries. In the present study, highly purified HCG was labeled with iodine-131 and used for the imaging of luteinized rat ovaries. Excellent images were obtained. The accumulation of radioactivity in the ovaries was completely inhibited by previous administration of excess unlabeled HCG. Time-course studies show that radioiodinated HCG was rapidly accumulated within 1 hr and largely cleared from the ovary at 24 hr. These results show the potential for imaging of endocrine target tissues by hormone-receptor binding, and the possibility of quantitation of hormone receptors in tissues under pathological conditions.


Tumorgenicity of herpesvirus hominis type 2-transformed cells (line 333-8-9) in adult hamsters. Inoculation of adult hamsters with greater than 1.5 X 10(2) Herpesvirus type 2 (HSV-2) transformed hamster embryo fibroblasts induced rapidly growing tumours at the site of inoculation, in more than 50 per cent of animals. The tumours were examined by light and electron microscopy, and were histologically anaplastic fibrosarcomas. Lung metastases were frequently found in tumour-bearing animals; these were usually only seen microscopically, and were cytologically similar to the primary tumours. The nature of tumour development showed similarities with naturally occurring tumours of man, and HSV-2 tumour is probably a better model of human cancer than other virus-induced tumours studied in the past.


Input-output relations in the pathway of recurrent inhibition to motoneurones in the cat. 1. The output from Renshaw cells caused by a phasic motor volley was investigated when these neurones were submitted to a background firing secondary to a tonic motor discharge elicited by a repetitive stimulation of muscle group I afferents. It was invariably found in individual Renshaw cells that tonic excitation produced an increase in the additional ouput caused by the phasic motor volley. The curves displaying this increase exhibited a significant 'jump' when the output resulting from the combined tonic and phasic motor discharges ranged between 2 and 5 spikes during the first 10 msec following the phasic volley. 2. The whole pool of Renshaw cells was also considered by assessing the amount of recurrent inhibition in motoneurones following a phasic motor volley. Similarly it was found that additional recurrent inhibition elicited by a phasic motor volley was enhanced when the Renshaw cells received a tonic excitatory input. 3. The Renshaw cell discharges elicited by stimulation of two different nerves were compared when a conditioning stimulus was previously applied to only one of them. The results strongly suggest that a preceding volley caused a decrease in synaptic efficacy at the terminals of the recurrent collaterals. 4. The firing produced by current injected through the recording micro-electrode was investigated in one intracellularly recorded Renshaw cell. It was found that the current-frequency curve was linear for 'steady-state' firing but displayed a clearcut sigmoid shape for the earliest intervals before the final adaptation. 5. It is demonstrated that this sigmoid input-output relation in individual Renshaw cells is sufficient to explain how the controls acting on these neurones may change the gain in the recurrent pathway when Renshaw cells are fired by a phasic motor discharge. When Renshaw cells are fired by a longlasting tonic motor discharge the linear input-output relation in individual cells should not cause any modifications of the gain in the recurrent pathway. A change in this gain secondary to the effects of segmental and supraspinal control systems is, however, possible if the motor discharge creates a subliminal fringe within the pool of Renshaw cells.


An electrophysiological analysis of the actions of prostaglandin on neuromuscular transmission in the guinea-pig vas deferens. Effects of prostaglandins (PGs) on the neuromuscular junction of the guinea-pig vas deferens were investigated by the micro-electrode and double sucrose gap methods. 1. PGE1, PGE2 or PGF2 alpha (10(-10) to 10(-7) g/ml.) did not change the membrane potential, membrane resistance and electrical threshold required to produce the action potential. 2. PGs markedly suppressed the amplitude of excitatory junction potential (e.j.p.) but the facilitation phenomena seen with repetitive stimulation were not affected. PGs did not change the amplitude and frequency of the miniature e.j.p. 3. The action potential recorded from the small nerve bundle with the aid of an extracellular micro-electrode method was not affected by PGS. 4. The amplitude of the e.j.p. was dependent on the external concentration of Ca. When plotted against [Ca]o on a double log scale, the above relation yielded a straight line with a slope of 1.0. Application of PGs resulted in a parallel shift of this relation to the right. 5. From the concentrations of [Ca]o to produce a given e.j.p. in the absence and presence of various concentrations of PGE1, the linear relation was observed from double logarithmic plot between [[Ca]o ratio -1] and [PGE1]o. However the slope of the line was too small (-0.37) to indicate a simple competition between [PGE1]o and [Ca]o. 6. Indomethacin (10(-6) g/ml.) had no effect on the amplitudes of e.j.p. produced by repetitive stimulation. 7. These results indicate that a low concentration of PGs interact with [Ca]o at the activated nerve terminals, but that there is no modification of the actions of Ca in the nerve terminals required for release of chemical transmitter.


Synthesis and biological properties of 2-, 5-, and 6-fluoronorepinephrines. 2-Fluoro-, 5-fluoro- and 6-fluorodimethoxybenzaldehydes were prepared by photochemical decomposition of the corresponding diazonium fluoroborates. The aldehydes were converted to the cyanohydrin trimethylsilyl ethers, which, in turn, were reduced to the dimethoxyphenethanolamines. Boron tribromide demethylation afforded the racemic ring-fluorinated norepinephrines. An alternate route, using the dibenzyloxyfluoroaldehyde, was also used to prepare 6-fluoronorepinephrine. The fluorine substituent markedly increases the phenolic acidities of these analogues. The biological properties conferred upon norepinephrine by the fluorine substituents in peripheral and central adrenergically responsive systems clearly demonstrate that 2-fluoronorepinephrine is a nearly a pure beta-adrenergic agonist, while 6-fluoronorepinephrine is an alpha-adrenergic agonist. 5-Fluoronorepinephrine retains both beta- and alpha-adrenergic agonist properties. Receptor-binding studies with specific radiolabeled ligands indicate that the specificity conferred by the site of fluorine substituents results from a change in the affinity of these analogues for the alpha- and beta-adrenergic receptors.


Derivatives of 1,3-disubstituted 2,4(1H,3H)-quinazolinediones as possible peripheral vasodilators or antihypertensive agents. A series of 1,3-disubstituted 2,4(1H,3H)-quinazolinediones was prepared from the 3-substituted 2,4(1H,3H)-quinazolinediones by treatment with sodium hydride and the desired alkyl halide in xylene. These compounds showed varying degrees of vasodilation and antihypertensive activity without significant blockade of alpha-adrenergic receptors. 1-[3-(N,N-Dimethylamino)propyl]-3-[3-(4-phenyl1-piperazinyl)propyl]-2,4(1H,3H)-quinazolinedione, which was selected for further studies, was more potent than papaverine in inducing vasodilation and induced a prolonged decrease in systolic blood pressure of hypertensive rats upon oral administration.


Mitochondrial antibodies--heterogeneity and effects on mitochondrial respiration. Sera containing antimitochondrial antibodies (MTA) were tested for binding to intact mitochondria, sonic fragments (SMP), Complex I + III and to oligomycin sensitive ATPase (OS-ATPase) from bovine heart by indirect immunofluorescence. Antigens capable of binding to MTA were present in mitochondria and its fragments tested. Maximum binding was observed with SMP. It appears that one or more antigen binding sites are present on the matrix side of the inner mitochondrial membrane or some location exterior to the inner membrane. Normal human serum or sera containing MTA did not effect the respiration of intact mitochondria or sonic particles. However, NADH-cytochrome c reductase activity of complex I + III was enhanced by 10-60% by sera containing MTA antibodies.


Isolation and characterization of a new simian foamy virus serotype from lymphocytes of a Papio cynocephalus baboon. A new type of syncytium-forming (foamy) virus was isolated from peripheral lymphocytes of a Papio cynocephalus baboon after cocultivation with a rhesus monkey cell line. The isolate was characterized by cytopathogenic effect, host range, electron microscopy, reverse transcriptase activity and physical and chemical properties. Seroneutralization tests indicate that the new isolate is antigenically distinct from the other nine known simian foamy virus serotypes.


Studies on lithium transport across the red cell membrane. VI. Properties of a sulfhydryl group involved in ouabain-resistant Na+-Li+ (and Na+-Na+) exchange in human and bovine erythrocytes. The reactivity of the SH-group essential for ouabain-resistant Na+-Li+ (and Na+-Na+) exchange and its location within the membrane are studied on human and beef erythrocytes and beef red cell ghosts. N-ethylmaleimide (NEM), 1,6-hexane dimaleimide, and iodoacetamide can induce an irreversible, partial inhibition of Na+-Li+ exchange in erythrocytes of the two species. The development of the inhibition due to the alkylating agents is greatly accelerated by external Na+ and Li+. The inhibition takes 3 min (NEM) and 60 min (iodoacetamide) to come to completion in isotonic Na+ media, but is hardly detectable in choline+, K+ or Mg2+ media. The transport site of the exchange system and the site promoting NEM binding exhibit similar affinities for external Na+. The impermeable, monofunctional glutathione derivative of 1,6-hexane dimaleimide does not inhibit Na+-Li+ exchange. The mercurials PCMBS, PCMB, and Hg2+ inhibit Na+-Li+ exchange in beef, but not in human erythrocytes. The inhibitory action of PCMBS, being slightly accelerated by external Na+, is fully reversed by penetrating thiols such as 2-mercaptoethanol, whilst glutathione, an impermeable thiol, is ineffective. Pretreatment with PCMBS affords partial protection from the irreversible inhibition caused by NEM. Oxidation with copper orthophenanthroline inhibits Na+-Li+ exchange only when performed in the presence of penetrating thiols such as 2-mercaptoethanol. It is concluded that the SH-reagents studied inhibit Na+-Li+ exchange by modifying an essential SH-group of a membrane protein in such a way that the turnover number of the exchange system is reduced. This SH-group is separated from both the red cell exterior and interior by a penetration barrier and seems to be distinct from the cation binding site. The action of external Na+ and Li+ in promoting the reaction of alkylating inhibitors is interpreted to result from a conformational change of the transport protein induced by the binding of external Na+ or Li+.


An improved method for estimating sequence divergence between related DNAs from changes in restriction endonuclease cleavage sites. We have developed a theory to estimate the degree of sequence divergence between related DNAs from the comparison of restriction endonuclease recognition sites. Two major improvements have been made upon a similar method reported by Upholt (1977). First, the most probable value is calculated by the collective use of all available data. This reduces intrinsic statistical error and extends the analyzable range of sequence divergence. Second, all variables are redefined so that they have strict mathematical implications. This corrects a serious error arising from the misinterpretation of the meaning of the fraction of conserved cleavage sites. With this refined method, sequence divergence between rat and mouse mitochondrial DNAs (mtDNAs) was calculated to be about 25% substitutions/nucleotide, which is in good agreement with the DNA-DNA hybridization data obtained by Jakovcic et al. (1975). It was also estimated that the three types of rat mtDNAs differ from one another by 0.3 approximately 1% of total base pairs. These values are 2 approximately 5 times smaller than those obtained with the conventional method.


Trophoblastic neoplasia in an African urban population. A clinical study of trophoblastic neoplasia in a Nigerian population in Lagos over a four-year period is reported. A high incidence of one in 379 deliveries for benign trophoblastic tumor and one in 846 deliveries for malignant tumor was found. Seventeen percent of benign trophoblastic tumors in this series progressed to the malignant type, but malignant trophoblastic tumor was preceded by the benign type (hydatidiform mole) in 46 percent of cases. The anterior vaginal wall is a common site for metastases of malignant trophoblastic neoplasia and, in one patient, the lesion progressed further to form a vesicovaginal fistula. While the management of benign disease was conservative, all cases of malignant trophoblastic neoplasia received chemotherapy.


Effect of linoleic acid hydroperoxide on liver microsomal enzymes in vitro. Rat liver microsomes incubated with linoleic acid hydroperoxide (LAHPO) lost cytochrome P-450 specifically among the enzymes of microsomal electron transport systems. The loss of cytochrome P-450 content and glucose-6-phosphatase activity by LAHPO was accompanied by an increase in malondialdehyde (MDA) production. Turbidity of microsomal suspensions was decreased with increasing MDA production, but not proportionately. Diethyldithiocarbamate (DTC), N,N'-diphenyl-p-phenylenediamine and alpha-tocopherol inhibited almost completely the LAHPO-induced MDA production of microsomes, however no perfect protection against the loss of cytochrome P-450 content and glucose-6-phosphatase activity was observed. The decrease of microsomal turbidity by LAHPO was little affected in the presence of DTC. Purified cytochrome P-450 was destroyed by LAHPO, with minimal protection by the compounds described above. These results suggest the possibility that the loss of microsomal enzyme activities during lipid peroxidation may be attributed largely to a direct attack on enzyme proteins by lipid peroxides rather than indirectly to a structural damage of microsomal membranes resulting from peroxidative breakdown of membrane lipids.


Radioimmunoassay of methionine enkephalin and interference by brain factor of immunoreactivity and opiate receptor binding activity. We developed a radioimmunoassay of methionine-enkephalin (met-enk) which was found to be highly specific to the peptide and 1 pmol was the detection limit. Interference by the following peptides was practically negligible; leucine-enkephalin, LAP- or CNBr-treated met-end, alpha- and beta-endorphins. Morphine and its congeners were totally inert. Measurement of met-enk in rat brain P2 fraction revealed besides met-enk, the presence of the factor which cross-reacted with the antiserum and also appeared to bind with the brain opiate receptor. Though the nature of the factor has not yet been characterized, it is suggested that for qualitative assessment of the tissue level of met-enk, a method to selectively inactivate either the peptide or the factor should be developed first.


Effects of chronic treatment with captopril (SQ 14,225), an orally active inhibitor of angiotensin I-converting enzyme, in spontaneously hypertensive rats. The effects of hydralazine (3 mg/kg) and the angiotensin I-converting enzyme (ACE) inhibitor captopril (SQ 14,225) (100 mg/kg) on mean arterial blood pressure, plasma renin activity, urinary volume and urinary Na+,K+, and aldosterone concentrations were examined in spontaneously hypertensive rats of the Okamoto and Aoki strain (SHR) after oral daily dosing for 2 weeks, 3 or 6 months. Captopril caused progressive cumulative reductions in blood pressure resulting in normalization of pressure after 6 months of dosing. Hydralazine also significantly reduced blood pressure but not to the level of normotensive rats of the Wistar-Kyoto strain (WKY). Reductions in heart size paralleled the changes in blood pressure, normalization of cardiac hypertrophy occurring after captopril but not hydralazine. Plasma renin activity increased approximately 2-3 fold after hydralazine and 15-fold after captopril. Neither hydralazine nor captopril had any consistent effects on 24-hr urine volume, urinary Na+,K+ or aldosterone excretion. These results indicate that chronic inhibition of ACE with captopril induces normalization of blood pressure in SHR, a normal-renin model of hypertension.


Failure of dibutyryl and 8-bromo-cyclic GMP to mimic the antagonistic action of carbachol on the positive inotropic effects of sympathomimetic amines in the canine isolated ventricular myocardium. Effects of carbachol, dbcGMP and 8-bromo-cyclic GMP on the positive inotropic actions of sympathomimetic amines and dbcAMP were studied on the canine isolated right ventricular myocardium. Carbachol alone did not substantially change the developed tension of the muscle, but did markedly shift the dose response curve for isoprenaline on the developed tension to the right and depressed the maximal response to the drug in a concentration dependent manner. The positive inotropic action of phenylephrine was affected by carbachol in the same manner as that of isoprenaline. DbcGMP in concentrations of 10(-3) M and higher produced a significant increase in the developed tension. The positive inotropic action of dbcGMP was partly inhibited by a beta-adrenoceptor blocking agent, pindolol. In the presence of dbcGMP, isoprenaline produced an action similar to that seen in the control experiment, but this action was not maintained on such a steady state level as in the control. This rapid decline of the effect of isoprenaline in the presence of dbcGMP was prevented by adding ascorbic acid to the organ bath. The positive inotropic actions of phenylephrine and of dbcAMP were not substantially affected by dbcGMP. 8-Bromo-cyclic GMP in a concentration of 10(-4) M did not change either the basal developed tension or the positive inotropic actions of noradrenaline and of isoprenaline. The present results indicate that these cyclic GMP derivatives are not able to mimic the antagonistic action of cholinergic stimulation on the positive inotropic action of adrenergic stimulation on the positive inotropic action of adrenergic stimulation on the canine ventricular myocardium.


Effects of smooth muscle stimulants on 1-isoprenaline-beta-adrenoceptor interaction in taenia from guinea pig caecum. An increase in intracellular cyclic AMP levels induced by 1-isoprenaline (1-Iso) was significantly depressed in coexistence with BuTMA or histamine, but not with KCl or BaCl2. An analysis of steady state data for d,1-[3H]-isoprenaline binding to single cells of taenia from guinea pig caecum in the presence of histamine or KCl by Scatchard plots was performed. The affinity for 1-Iso for beta-adrenoceptor sites was decreased in the presence of histamine but not in the presence of KCl. These results suggest that the interaction of 1-Iso with the beta-adrenoceptors is interfered with by histamine and butyltrimethylammonium, for which the specific receptors were postulated, but not with KCl and BaCl2, nonspecific smooth muscle stimulants. These phenomena demonstrate the heterotropic negative cooperatively among the heterogeneous receptors.


Effect of colchicine on steroid secretion from rat adrenal gland. To determine the influence of colchicine on adrenocortical function in vivo, we gave significant amounts subcutaneously to rats and observed the distribution to the anterior pituitary and adrenal glands. Corticosteroids in the serum and adrenal gland were increased remarkably by the administration of colchicine in a dose-dependent manner. Maximum elevation was achieved around 2.5 hours after the injection and continued for at least 4 hours. Maximum levels produced by the injection of 0.1 U/rat of adrenocorticotropic hormone, the latter producing temporary increases in corticosteroids in the serum and adrenal gland. A significant augmentation of corticosteroids in the serum and adrenal gland resulted when colchicine was administered 60 minutes prior to the injection of adrenocorticotropic hormone. The pattern of intracellular distribution of corticosteroids in the adrenal glands of rats given colchicine was the same as that seen in rats given adrenocorticotropic hormone. In the in vitro experiment, only high concentrations of colchicine such as 10(-3) M added to the incubation system of the adrenal quarters suppressed the release of corticosteroids from the adrenal gland. Thus, the continuous stimulation of the synthesis and secretion of adrenocortical hormone seen with the administration of colchicine may explain why this drug is effective in the treatment of gouty arthritis.


[Significance of virus-specific antibodies to EBV-antigens in patients with nasopharyngeal carcinoma (author's transl)]. Sera collected from patients with nasopharyngeal carcinoma, patients with other head and neck neoplasms, patients with infectious mononucleosis (IM), and healthy donors were titrated for antibodies (IgM, IgG, IgA) against Epstein-Barr-Virus (EBV) specific antigens (VCA, EA, EBNA) by indirect and by antikomplementary immunfluorescence. NPC-patients develop significantly high anti-EBV titers of IgG and IgA. In contrast patients with neoplasms (mainly carcinomas) arising in sites of the head and neck other than the nasopharynx revealed a lower incidence of high titers. Our results emphasize the remarkable predominance of IgA-antibodies to VCA and EA in NPC-patients and shows that Europeans to not differ in that respect from Asian patients. The significance and implications of these findings are discussed.


Replication of the colicin E1 plasmid in extracts of Escherichia coli: uncoupling of leading strand from lagging strand synthesis. The replication of the ColEl plasmid was studied in extracts from E. coli dnaG mutants. It was found that the synthesis of the complementary strands of ColEl DNA can be carried out in these extracts in two consecutive steps: (1) synthesis of the leading L strand independent of the dnaG function, and (2) synthesis of the lagging H strand depending upon addition of wild-type dnaG protein. In contrast to L strand synthesis, the latter reaction is insensitive to rifampicin and novobiocin. Both synthetic pathways are however blocked by antiserum directed against dnaB protein. This indicates an additional role of the dnaB protein in duplex DNA replication besides assisting the dnaG protein in the priming of lagging strand synthesis. The T7 gene-4 protein acting in conjunction with T7 DNA polymerase can substitute for both the function of the dnaB and dnaG protein. It is concluded that plasmid replication proceeds by a semi-discontinuous mechanism.


[Salmonellosis in children in Ivory Coast, (A study on 69 cases) (author's transl)]. Salmonellosis are still endemic in Ivory Coast. In 1975 they were responsible for 1,75 p. 100 of the admittances in the pediatric department of Treichville University Hospital. In children, the conventional distinction between major and minor salmonellosis is not always obvious. Interpretation of Widal sero-diagnosis is often difficult and awkward. Chloramphenicol generally leads to satisfactory evolution. Salmonellosis are still a problem for Public Health Service. Immunization, is the only effective prophylaxis, but is not carried out yet in Ivory Coast.


In vitro insertion of the lambda attachment site into the plasmid RP4. The region of the phage lambda chromosome containing the attachment site (P.P') and the genes int and xis, excised by the action of endonuclease R.EcoRI, has been inserted into the unique site for that enzyme on the promiscuous conjugative plasmid, RP4, generating the recombinant plasmid RP4att lambda. Transformants containing the hybrid plasmid were recognised by their ability to allow efficient lysogenization by phage lambda b2 (Weil and Signer, 1968; Echols et al., 1968) containing the mutant attachment site delta. P'. The construction and properties of the hybrid plasmid RP4att lambda are described.


DNA supercoiling and transcription in Escherichia coli: influence of RNA polymerase mutations. Coumermycin A1, a specific inhibitor of DNA gyrase, differentially changes the spectrum of proteins synthesized in wild type E. coli cells but has no effect on the protein spectrum in mutant cells with coumermycin-resistant DNA gyrase. The rpoB265 mutation affecting RNA polymerase decreases the coumermycin A1-sensitivity of bacteria while the rpoC3 mutation increases it. The interaction of wild type and mutant RpoB265 RNA polymerases with ColEl plasmid DNA in vitro is differently affected by DNA supercoiling. No such differences are observed in the case of RpoC3 RNA polymerase. The results suggest that template supercoiling may have a substantial effect on transcription in vivo, an effect which, in some cases, depends on the properties of RNA polymerase.


A cleavage map of the TOL plasmid of Pseudomonas putida mt-2. A cleavage map of the TOL plasmid pWWO has been determined for the restriction endonucleases HindIII and XhoI. A number of techniques were employed including (i) digestion of purified cleavage products with a second enzyme; (ii) hybridisation of purified XhoI fragments to Southern blots of HindIII digest products and (iii) analysis of a number of deletion mutants.


Absence of relationship between UV-induced reversion frequency and nucleotide sequence at the CYC1 locus of yeast. The UV induced mutation frequency of a given base pair located at different sites within the CYC1 gene of Saccharomyces cerevisiae was found to vary by more than fifty fold, indicating the existence of hotspots and coldspots typical of those found in other organisms. We were unable, however, to find any feature of the nucleotide sequence at or near the sites of mutation that explains this variability. These and other data suggest that hotspots are not located within regions particularly susceptible to the formation of premutational lesions. More probably the variation in mutability depends on differences in the activity of enzymes responsible for producing mutations, though the reasons for these differences are not understood and may depend on factors not directly related to nucleotide sequence.


A restriction enzyme cleavage map of Tn5 and location of a region encoding neomycin resistance. This paper reports a cleavage site map of Tn5 for restriction enzymes BamHI, Bg/I, Bg/II, Hind II, HindIII, HpaI, Sa/I, Aval, SmaI, XhoI, PstI, PvuII, HaeII and HaeIII that was determined by the analysis of restriction enzyme cleavage patterns of ColEl, two independent ColEl::Tn5 plasmids, and a ColEl::Tn5 deletion derivative. Ba/I, EcoRI, KpnI, and PvuI do not cleave Tn5. Construction and analysis of in vitro-generated deletions of a ColEl::Tn5 plasmid limit the sequences encoding neomycin resistance to a 1500-base-pair-long segment of Tn5. Insertion of DNA at a Bg/II site within this segment results in loss of the neomycin resistance phenotype. Since this Bg/II site lies in an inverted repeat region, sequences within this repeat seem to be involved in the expression of neomycin resistance.


Reactivity of Paul-Bunnell type heterophile antibody in sera from infectious mononucleosis patients with the surface of lymphoid cells carrying Epstein-Barr virus genomes. The possible presence of Paul-Bunnell (PB) antigen on Epstein-Barr virus (EBV)-transformed lymphocytes were investigated. Of 23 EBV genome-positive lymphoblastoid cell lines tested all but one absorbed PB type antibody from the serum of an infectious mononucleosis patient. The one EBV-negative B cell line tested did not absorb the heterophile antibody. PB antibody, purified by an immunoadsorbent procedure using beef cell antigen, reacted with the EBV producer P3HR-1 cell line in an indirect membrane immunofluorescence test and was shown to be IgM antibody. Titers of heterophile agglutinin and reactivity with the cell surface were reduced to the same degree by absorption with beef cell antigen but not with guinea pig kidney antigen. PB antibody was distinct from IgM antibody against the EBV-determined membrane antigen, since the latter was not absorbed by beef cell antigen. PB antibody was also reactive with other EBV-positive B cell lines (QIMR-WIL, NC-37, and Raji) which were free of surface IgM. No reaction occurred with the nonproducer P3HR-1 line, a null cell line, and two T cell lines. The results suggest the presence of PB antigen on most EBV-transformed B lymphocytes, and its appearance in each of the transformed lymphocytes of patients with acute infectious mononucleosis.