Observation on intranuclear crystal and nucleolar size at different stages of cell differentiation in the midgut epithelium of several insects. Based on an inverse size relationship between nuclear crystal and nucleolus in different cells it has been postulated by several authors that the crystal develops from nucleolar materials. The purpose of the present paper is to investigate the validity of this argument. Intranuclear proteinaceous crystals appear in differentiating midgut cells of Gyrinus marinus and Tenebrio molitor. In an autoradiographic study we have previously demonstrated in these two species that the crystals do not develop from nucleolar materials. However, an inverse relationship with regard to size is observed between these 2 structures during the cell differentiation: the cross-sectional area of the nucleolus decreases when the cross-sectional area of the crystal increases. But a decrease in size of the nucleolus is also observed during the differentiation of the midgut cells of Gyrinus natator where the crystals are not present. Consequently an inverse size relationship cannot be a sufficient argument to postulate that intranuclear crystals and nucleoli are interconvertible structures; decrease in size of the nucleolus is not related to development of the intranuclear crystal.


Determination of conjugated and esterified estrogens in pharmaceutical tablet dosage forms by high-pressure, normal-phase partition chromatography. A high-pressure, normal-phase partition chromatographic method for the identification and determination of conjugated and esterified estrogens in pharmaceutical tablet dosage forms is described. The method is based on the separation of the estrogen sulfate esters on a conventional diatomaceous earth-H20 column, followed by HCI-methanol hydrolysis, and finally a chromatographic separation on a chemically bonded ether (ETH-Permaphase) column and measurement. Mobile phases found useful were combinations of 2-propanol and n-heptane. Equillin and -dehydroestrone, a structurally related isomer, not separately determined in the proposed method, are determined as a sum by performing a chromatographic study of their corresponding 2, 4-dinitrophenylhydrazine derivatives. Studies have indicated several estrogen derivatives can be rapidly formed and then separated on the same column, providing a useful qualitative analysis scheme. Commercial tablet dosage forms were analyzed for conjugated and esterified estrogens and the results are presented.


Hypertensive virilizing adrenal hyperplasia with minimal impairment of synthetic route to cortisol. One of the first described cases of hypertensive virilizing adrenal hyperplasia (VAH) (Pediatrics 8: 805, 1951) has been followed from age 2 1/2 until age 26. Blood pressure as an infant was 150/90, and at age 25 was 220/160. During childhood the patient was lost to follow-up for prolonged periods, and received no therapy from age 20 to 25. At this time 24 h urinary excretion of 17-ketosteroids was 89 mg; tetrahydro 11-deoxycortisol (tetrahydro S), 47 mg and pregnanetriol 5.7 mg. Hourly measurements of several plasma steroids utilizing sephadex LH 20 chromatography and competitive protein binding were made during 24 h; concentration ranges were made during 24 h; concentration ranges were as follows (mug/100 ml): 11-deoxycortisol 8-40; cortisol 0-48; corticosterone 0-15; deoxycorticosterone 1-18. Plasma cortisol, especially showed a significiant morning impairment, but reached normal and even markedly elevated levels during the day and early evening. Urinary cyclic AMP per 24 h ranged from 5.3 to 11.6 n mol/mg creatinine before therapy, and was 1.9 n mol after therapy. The results suggest either the formation of an alternate pathway to cortisol synthesis, or the existence of a form of VAH with two independent 11-B hydroxylating systems, exhibiting only minimal impairment of the synthetic route to cortisol. The latter would support the presence of two independent 11-B hydroxylating systems in the normal human adrenal. This has been suggested by Zachmann et al. (J Clin Endocrinol Metab 33: 501, 1971) to be true in infancy. Our observations on an adult indicate that these two systems may not be transitory, but persist into adulthood.


Radioimmunoassay of 18-hydroxy-11-deoxycorticosterone in plasma. A method is described for the radioimmunoassay of 18-OH-DOC using antibodies generated in rabbits against the carboxymethoxime derivative coupled to bovine serum albumin. The procedure uses 4 ml of plasma with intra and interassay variations of 8 and 9% respectively. Standard 18-OH-DOC added to plasma from an adrenalectomized patient gave a regression equation, Y=0.974X+/-2.210 and a correlation coefficient of 0.999. The only cross reacting steroid, 18-OH-B which may lead to falsely high levels is removed by a single thin layer chromatographic step. Blood levels in normal subjects agree closely with those calculated indirectly by metabolic clearance and secretion rate measurements. ACTH stimulation produced an 18-fold increase in plasma concentration while dexamethasone suppression decreased levels 3-fold. Four hours in the upright position resulted in a decreased plasma concentration while aldosterone increased. No significant response to dietary sodium restriction could be demonstrated.


Chemical characterization of ectopic ACTH purified from a malignant thymic carcinoid tumor. ACTH was purified from thymic tumor associated with ectopic ACTH production by gel filtration and ion exchange chromatography. Amino acid composition and C- and N-terminal analyses indicated that this tumor ACTH was comprised of the 2-38 sequence of authentic pituitary ACTH. The observations suggest that the mode of cleavage of this tumor ACTH from its precursor is different from that of pituitary ACTH.


Periodic remission in Cushing's disease with paradoxical dexamethasone response: an expression of periodic hormonogenesis. A patient with Cushing's disease due to a chromophobe adenoma was studied for 243 days before pituitary surgery and evidence for periodicity in cortisol steroid production was found with cycles occurring every 85.8 days (peak-to-peak length), associated with laboratory remissions and paradoxical response to dexamethasone. The autonomy of ACTH secretion was suggested by the nonresponsiveness to repeated lysine-vasopressin stimulation tests and lack of increase in urinary 170HCS following metyrapone. A distinct response of the hyperplastic glands (as demonstrated by percutaneous adrenal venography) was obtained on several B1-24 corticotropin stimulation. The patient's hypercortisolism disappeared following removal of the chromophobe adenoma through transphenoidal hypophysectomy.


Gonadotropin output in congenital adrenal hyperplasia before and after adrenal suppression. Basal release of gonadotropin and the response to an infusion of 100 mug of synthetic luteinizing hormone releasing hormone (LRH) were studied in a teenage girl with congenital adrenal hyperplasia (CAH). The initial study was done during a period of poor adrenal suppression, and second study was done after adequate adrenal suppression was achieved. To assess adrenal function, circulating levels of adrenal steroid hormones were evaluated continuously over a 24 h period. During the period increased production of adrenal androgens, the pattern of gonadotropin release was that of a prepubertal child. After 3 months of adrenal suppression the pattern of gonadotropin secretion was similar to that of a normal girl in mid-puberty. This demonstrates the rapid change from prepubertal to pubertal gonadotropin dynamics in a teenage patient following adequate suppression of androgens from the adrenal.


Evaluation of the sodium polyanethol sulfonate disk test for the identification of Peptostreptococcus anaerobius. The previously reported sodium polyanethol sulfonate disk test for the identification of Peptostreptococcus anaerobius (Graves et al., 1974) was evaluated, with modifications. Three bands of brucella agar, three inoculum sizes, and two inoculum sources were compared. Nine stock cultures of P. anaerobius (eight normal flora isolates and ATCC 27337) and 16 fresh clinical isolates were used. All cultures of P. anaerobius showed inhibition zones of 12 to 30 mm in diameter, regardless of test conditions. Out of 103 clinical isolates of other species of anaerobic gram-positive cocci tested, only two had an inhibition zone size in this range (one P. micros of 11 studied had a zone of 12 mm and one P. prevotii of 14 studied had a zone of 16). The test had an overall accuracy of 98% in the identification of P. anaerobius from clinical specimens. Since P. anaerobius accounts for one-fifth to one-third of all anaerobic gram-positive cocci encountered in clinical specimens, this simple and rapid technique can be very useful for presumptive identification.


Screening tests for hepatitis B antigen and antibody in two colleges of education and studies on the relationship between nonspecific positive antibody tests and EB virus infection. Sera from 627 students entering Colleges of Education between 1969 and 1972 were tested for hepatitis B surface antigen and antibody. One was found positive for antigen, none for antibody. Six for 15 positive Hepanosticon results and two positive Hepatest results occurred in sera which also gave positive heterophil antibody tests indicative of current or recent EB virus infection. One of these six sera was still positive in the Hepanosticon test after one absorption, and one of two Hepatests gave no positive reaction with the control cells. Eleven of 14 sera from cases of infectious mononucleosis gave positive Hepanosticon results and two were still positive after one absorption. Seven were positive in the Hepatest and only three of these were positive with the control cells. The control tests in the Hepanosticon and Hepatest do not clearly identify all false positives due to Paul Bunnell antibody. It is suggested that when a positive result in a passive haemagglutination test can be removed by absorption or if positive after absorption cannot be confirmed by other tests for hepatitis Bs antigen, the patient from whom the serum specimen was taken should be investigated for indications of current EB virus infection.


Malignant fibrous histiocytoma with diffuse spinal nerve involvement. Two cases of malignant fibrous histiocytoma in which paraplegia was a major feature are presented. The histological features of the tumours are discussed with particular emphasis on the neural involvement. A brief review of the literature on the subject is included.


Hepatic lesions in patients treated with synthetic anabolic steriods. Hepatic abnormalities are described in three patients who received synthetic anabolic steroids. A child with Franconi's anaemia was treated for four years and at necropsy the liver showed generalized hyperplasia, hyperplastic nodules, and a benign hepatoma. Two adults received only three months' therapy with synthetic androgens; in one there was generalized hepatic hyperplasia and in the other widespread nodular hyperplasia. It is suggested that anabolic steroids may induce tumours through intermediate hyperplastic lesions, a sequence similar to that seen during tumour induction by carcinogens in experimental animals.


Chemotherapeutic compounds and Acanthamoebae from eye infections. The amoebicidal and amoebistatic action in vitro of 24 compounds was tested on two strains of Acanthamoeba, A. polyphaga and A. castellanii, isolated from eye infections in this country. For comparison, the Ryan strain of A. castellanii and Naegleria gruberi L-1 were also examined. Nine compounds showed sufficient activity to merit further consideration, ie, acriflavine, proflavine, hydroxystilbamidine isethionate, paromomycin, miconazole, amphoterin, neomycin, polymyxin, and the last two combined in Neosporin.


Measurement of heterophil antibody and antibodies to EB viral capsid antigen IgG and IgM in suspected cases of infectious mononucleosis. The EBV IgG titres in acute and convalescent specimens from 97 cases of infectious mononucleosis were compared with titres from acute and convalescent sera from 96 students with illnesses resembling infectious mononucleosis but without heterophil antibody, EB IgM or EB IgG seroconversion; and also with titres from 91 healthy students known to have had EB IgG antibody for at least six months. These titres were related to the titre of the Research Standard A.66/235 for infectious mononucleosis serum prepared by the National Institute for Biological Standards and Control. Serial sera were tested for heterophil antibody and EBVCA specific IgG and IgM from 61 university students with infectious mononucleosis. The period of persistence of heterophil antibody and EBV IgM after illness was outlined from the results of the tests. Single sera from 406 patients in hospital or general practice sent to the diagnostic laboratory for heterophil antibody tests were also tested for EBV antibodies without prior knowledge of the heterophil antibody result. The close agreement between heterophil antibody and EBV IgM results is shown. False positive EB IgM results were correlated with the presence of rheumatoid factor.


Cell-mediated immunity to herpes simplex virus in man. V. Antibody-mediated cell-dependent immune lysis of herpes virus-infected target cells. Thirty-four patients, subject to recurrent herpes labialis, have been studied. They have all been shown to have high serum levels of an antibody to HSV1. This antibody has the property of sensitizing HSV1-infected target cells to lysis by nonimmune effector lymphocytes. Of 23 subjects who gave no history of herpes labialis, only four had antibody demonstrable by this technique. The level of antibody remains essentially unchanged despite recrudescenes of herpes labialis in the susceptible subjects. Effector cell activity was present in all of these and other subjects tested except for two who were suffering from chronic lymphatic leukemia. We have positive evidence that the effector cells in this system are neither T cells nor macrophages. Additional evidence suggests that the effector cells may be "null" cells.


Decreased adrenergic responses in lymphocytes and granulocytes in atopic eczema. The physiologic and cyclic adenosine monophosphate (cAMP) response to beta adrenergic stimulation in lymphocytes and granulocytes was examined in atopic eczema. These cells were isolated by Ficoll-Hypaque gradient from 10 patients with atopic eczema, and their responses were compared to 10 normal subjects. In eczema, basal concentrations of cAMP were normal in both lymphocytes and granulocytes. Lymphocyte cAMP response in eczema was decreased both to epinephrine (10-5 M) and to isoproterenol (10-5 M) but normal to prostaglandin E1 (PGE1). It was also noted that the glycogenolysis response to isoproterenol was significantly less at 10-5 M in eczema, but the fall in glycogen was normal with PGE (10-5 M and 10-7 M). The inhibition of lysosomal enzyme release from granulocytes after zymosan stimulation was significantly less (p less than 0.01) in eczema with all concentrations of isoproterenol tested. There was also a decrease in cyclic AMP response to isoproterenol in the polymorphonuclear leukocytes. PGE1 inhibited lysosomal enzyme release and stimulated cAMP normally. In eczema, both lymphocytes and polymorphonuclear leukocytes have a decreased beta adrenergic response.


Effect of heat on foot-and-mouth disease virus (FMDV) in the components of milk from FMDV-infected cows. Foot-and-mouth disease virus (FMDV) survived in skim milk, cream and the pelleted cellular debris components of milk obtained from FMDV-infected cows after pasteurization at 72 degrees C for 0-25 min. Virus was recovered from whole milk of infected cows after that milk was heated at 72 degrees C. for 5 min. and from the skim milk component after it was heated at the same temperature for 2 min. Evaporation of the whole milk samples after they were heated at 72 degrees C. for 3 min. did not inactivate the virus, but evaporation of infected skim milk samples after they were heated at 72 degrees C. for 0-5 min. did inactivate the virus. FMDV survived in the cream component after it was heated at 93 degrees C. for 0-25 min.


Prevalent serotypes of Bordetella pertussis in non-vaccinated communities. In many countries, the prevalent serotypes of Bordetella pertussis have changed from a mixture of types 1,2,3 and 1,2 (organisms possessing antigen 2) to a predominance of type 1,3. The timing of the change in different countries is shown to be related to the introduction of mass-vaccination with material rich in antigens 1 and 2 but weak in, or devoid of, antigen 3. In several parts of the world, there have been outbreaks of type 1,3 infection in fully vaccinated children. Non-vaccinated communities in various parts of the world still show the pattern of serotypes which existed elsewhere before mass-vaccination. In order to avoid the disappointments experienced in the past, it is essential that pertussis vaccine for use in previously non-vaccinated communities, like that for any other country, should be rich in each of the three antigens.


The skin flora of the hemiplegic hand.

The skin flora of the hemiplegic hand. More organisms were found on the paralysed hands of hemiplegic patients than on their unaffected ones. However, both showed considerably higher bacterial counts than did geriatric patients without paralysis. An infection rate for Cl. welchi of 15% was found in the paralysed hands.


Age-specific incidence of neutralization antibodies of Herpes simplex virus. Sera of 1255 individuals from Novi Sad, varying in age from less than 1 month to 69 years, have been tested for neutralization antibodies to Herpes implex virus type 1. The eight newborns tested and 97% of the 507 adults were positive, with titres ranging from 1/4 to 1/256. The titres in newborns were significantly lower than the titres in adults. After birth the maternal antibodies declined rapidly and 94% of infants at the age of greater than 6 months and less than 2 years were negative. After the first year infants in Novi Sad start to acquire herpes-neutralizing antibodies actively, reaching a 50% incidence of positives between the 2nd and 3rd year of age. Age-specific incidence rates of herpes positives found in Novi Sad have been compared with those reported from Edinburgh, Freiburg i. Br. and Louisiana. Possible influences of several circumstances upon the incidence rate of positives detected by the neutralization test are discussed.


Pathogenesis of foot-and-mouth disease: the lung as an additional portal of entry of the virus. Donor cattle infected with foot and mouth disease (FMD) virus subtype O1 were used to expose experimental cattle. The pharyngeal virus growth and viraemia patterns after contact exposure were quite different from those obtained after intranasal inoculation and suggested that the lower respiratory tract might provide an additional portal of entry for the virus. A tracheotomy was performed on experimental cattle to let the respiration bypass the pharynx, followed by exposure to FMD virus by different routes. The results confirmed that FMD virus can enter the bloodstream via the lung, followed by haematogenic infection of the pharynx and other replication sites simultaneously. These observations led to further experiments in which the intravenous route of infection was used to study the interaction of virus growth in the phayrnx, in other sites, and in viraemia.


Pathogenesis of foot-and-mouth disease: clearance of the virus from the circulation of cattle and goats during experimental viraemia. Viraemia is an important aspect of the pathogenesis of infectious diseases, but the mechanisms of entry and removal of virus from the vascular system particularly in natural hosts are poorly understood. The results of this study showed that the clearance of foot and mouth disease virus (FMDV) from the circulation of cattle and goats followed the general rules for the clearance of inert particulate materials and other viruses from the circulation. High doses of infused FMDV were cleared less efficiently than low doses, probably as a result of a depletion of the reticulo-endothelial system by the higher doses. FMDV was cleared from the circulation of cattle at a considerably slower rate than from the circulation of goats, but in both species significant individual variation in clearance was observed. These results could explain individual as well as species variations relative to the onset and duration of viraemia.


The assessment in sheep of an inactivated vaccine of parainfluenza 3 virus incorporating double stranded RNA (BRL 5907) as adjuvant. The serological responses of conventionally reared sheep were compared after vaccination with inactivated parainfluenza 3 (PI3) virus incorporated in three different adjuvants. Inactivated PI3 virus with the double-stranded RNA, BRL 5907 in an oil emulsion was shown to stimulate higher serum antibody titres over the first 5 weeks after vaccination than virus with and without BCG emulsified in oil. The ability of this vaccine to protect specific pathogen-free lambs against challenge with PI3 virus was examined in a second experiment. In this experiment the vaccine stimulated virus neutralizing and haemagglutination inhibiting antibodies in the serum. After intranasal and intratracheal inoculation with PI3 virus at challenge, vaccinated lambs showed no clinical illness and virus isolation was confined, except in one lamb, to the first two days. In contrast, unvaccinated lambs developed respiratory disease and virus was isolated daily for 7 days after challenge.


Natural infection with influenza A (H3N2). The development, persistance and effect of antibodies to the surface antigens. A technique for estimating antibodies to the neuraminidase antigens of influenza A is described. The antibody response to the haemagglutinin and neuraminidase antigens of influenza A (H3N2) was studied in a boys' public school over the four-year period 1970--4. During this time there were two outbreaks of influenza A and the effect of antibody on the result of natural challenge was investigated. No boy who had homotypic neuraminidase antibody had clinical influenza.


Cytomegalovirus excretion in gnotobiotic pigs. Germ-free piglets were infected intranasally with porcine cytomegalovirus (PCMV) at 1 day (group A) or 3 weeks of age (group B). Viraemia and virus excretion by the nasal, pharyngeal and conjunctival routes was studied up to the time of death or to 12 weeks. Virus was also sought in tissues at death or at slaughter, as well as in a few urine samples. Viraemia was detected in group A between days 5 and 19 after infection and in group B between days 14 and 16 inclusive. The chief route of virus excretion was the nasal mucosa, followed by the pharynx and conjunctiva; the maximal duration of excretion by these routes was 32, 25 and 14 days for pigs of group A and 9, 7 and 4 days for group B. The quantity of virus was also greater in the former group, of which died of generalized PCMV infection. A viruria was demonstrated in 2 animals. Antibody detectable in indirect immunofluorescence (IIF) tests appeared towards the end of the third week, reaching maximal titres at 5 to 7 weeks after infection. The mean peak titre of antibody in group B was lower than in group A. Corticosteroid treatment at days 56--62 after infection resulted in some recrudescence of virus excretion, accompanied in group B by about a twofold increase in IIF antibody. PCMV was isolated in cultures of lung macrophages from 4 of 7 animals killed at about 12 weeks after inoculation.


Hypergammaglobulinemia in chickens congenitally infected with an avian leukosis virus. Significantly elevated (2- to 5-fold higher than controls) serum levels of IgG were found in chickens congenitally infected with F42 strain of avian leukosis (ALV-F42) a subgroup A avian leukosis virus (ALV). A further increase in IgG levels in congenitally infected birds was found to be induced by injection of influenza virus in complete Freund's adjuvant(CFA). Serum immunoglobulin M (IgM) levels were not significantly elevated in ALV congenitally infected chickens except in those animals that had been injected with influenza virus in CFA. Hypergammaglobulinemia in ALV infected birds resulted only after congenital infection and not after infection of immunologically competent birds. Therefore this phenomenon appeared to have striking parallels with other persistent or chronic viral infections that have been previously described in mammals.


Depression of macrophage function by a factor produced by neoplasms: a merchanism for abrogation of immune surveillance. The possibility that macrophages mediate surveillance against the development of neoplasms has been reciving increasing support. The acquisition, by neoplastic cells, of the capacity to subvert macrophage function may be an important mechanism by which they escape destruction by the host and become established tumors. Indeed, animals implanted with syngeneic neoplasms developed depressed macrophage migratory ability in vivo and chemotactic responsiveness in virto. It therefore seemed plausible that neoplasms might be capable of producing inhibitors of macrophage function. The present report describes the identification of such a low molecular weight (6,000 to 10,000), heat-stable inhibitor of murine macrophage accumulation in vivo and chemotaxis in vitro. The inhibitor of macrophages was present in four different murine neoplasms, but not present in normal liver, spleen, or inflammatory exudate cells and did not affect PMN chemotaxis in vitro. When given with low numbers of neoplastic cells, the inhibitor increased both the frequency of tumor development and rate of tumor growth. By producing inhibitors of macrophage function, neoplasms may escape initial host surveillance mechanisms.


Characterization of Ia8 antigen, thy-1.2 antigen, complement receptors, and virus production in a group of murine virus-induced leukemia cell lines. Ia8, a cell membrane antigen controlled by gene(s) located in the I region of the H-2 complex, was found on 9 of 26 murine leukemia cell lines. Iaddition, 3 of the 9 Ia8-bearing lines had a membrane receptor for antigen-antibody-complement complexes. Six of 26 lines bore the Thy-1.2 antigen. Ia8 and Thy-1.2 antigens were mutually exclusive on the cell lines studied. The strain of virus used to induce the leukemia, the H-2 type of the cells, and the techniques of leukemia cell propagation all appeared to influence the antigenic characteristics of the cell lines obtained. Production of infectious murine leukemia virus in vitro and expression of leukemia virus-induced membrane antigens did not appear to correlate with the presence of I8 or Thy-1.2 antigens or with the H-2 type of the cells.


Independent effects of IgG and complement upon human polymorphonuclear leukocyte function. Particle ingestion by polymorphonuclear leukocytes (PMN) is promoted by cell surface recognition and binding of fragments of the third component of complement (C3) and Fc regions of certain immunoglobulin (IgG) molecules. In order to determine the influence of these specific ligandsurface membrane interactions upon other PMN functions, we have employed nonphagocytosable particles (serum-treated Sepharose beads) coated with fragments of C3 and/or IgG, and have investigated whether these provide a sufficient stimulus for the metabolic changes and degranulation that ordinarly accompany phagocytosis by PMN. Sepharose 4B activates complement in fresh normal serum and consequently is coated with fragments of C3 (confirmed by immunoelectrophoretic evidence of factor B and C3 conversion and by immunofluorescence). Adsorbed IgG could be removed from serum-treated Sepharose by boiling in 2 M NaCl without significantly influencing bound complement. We have found that normal human PMN recognize and adhere to Sepharose beads coated with fragments of C3 and consequently are stimulated to increase their oxidative metabolism (measured as superoxide anion generation). This PMN response occurred in the absence of IgG but could be amplified if this immunoglobulin was also present on the bead surfaces. Both adherence and metabolic stimulation could be blocked by treatment of the beads with F(ab)2 anti-C3. In contrast to metabolic stimulation, degranulation (selective extracellular release of lysosomal constituents) was observed only when PMN encountered both C3 fragments and IgG on the beads. This response could be blocked by treating beads with either F(ab)2 anti-C3 or F(ab)2 anti-IgG. These results indicate that cell surface stimulation of PMN is not an "all or none" phenomenon and that certain vital functions of these cells may be mediated or modulated independently by immunoglobulins and complement.


Lysis of tumor cells by antibody and complement. VII. Complement-dependent 86Rb release--a nonlethal event? The guinea pig hepatoma (line-1) treated with anti-Forssman antibody (TA) and GPC sequentially released 86Rb, 14C from 14C aminoidobutyric acid and failed to exclude trypan blue. Incubation of TA with fluid phase GPC for 1 min caused maximal 86Rb release; however, if the GPC was removed at this time, the cells were not subsequently killed. Using a number of naturally occurring human sera deficient in a complement component we have shown 86Rb release requires the binding of the complement components 1 through 8, but there was no absolute requirement for C9. Irreversible damage to the cell as measured by 14C AIB release or uptake of trypan blue required the complete sequence of complete sequence of complement components. These observations indicate that 86Rb release is not a relible indicator cytotoxicity.


Characterization of Ia antigens in mouse serum. Sera obtained from normal B10.BR mice were shown to inhibit selectively a specific anti-Ia alloantiserum. Partial purification of the Ia antigenic activity was accomplished by isolation of the high density lipoproteins from these sera by fractional precipitation with sodium phosphotungstate and MgCL2. Both H-2.23 and Iak antigens present in this high density lipoprotein fraction were completely adsorbed by rabbit anit-rat beta2-microglobulin immunoadsorbents, whereas specific anti-H-2.23 immunoadsorbents removed only the H-2 activity. These data deomnstrate that Ia antigens, like H-2 antigens in the sera of B10.BR mice are associated with high density lipoproteins and further suggest that both H-2 and Ia antigens are associated with a beta2-microglobulin-like molecule.


Effect of neutral red and light on Herpesvirus hominis type 1 in cell culture. Various concentrations of neutral red were added to monolayers of muscle-skin fibroblasts after adsorption of Herpesvirus hominis type 1. The concentration necessary to reduce plaque counts was found to be 10(-5.5) M. At the same time, the minimal toxic concentration of neutral red for muscle-skin fibroblasts was determined by the concentration that reduced the plaques of a challenge virus, vesicular stomatitis virus, that was applied after treatment with neutral red and light. The minimal toxic concentration was found to be 10(-5) M. Thus, the effective concentration of neutral red for H. hominis in tissue culture appears to be only slightly less than the minimal toxic concentration. The concentrations used for clinical trials in humans have been 10(3)-10(4) times this amount. Any observed efficacy of such treatment may be a reflection of cell toxicity.


Inter- and intralaboratory variability in antibiotic susceptibility tests with Pseudomonas aeruginosa and Enterobacteriaceae. A two-phase collaboratice study was designed for definition of the extent to which results of disk diffusion tests can be reproduced. Two currently recommended techniques, the Kirby-Bauer method and the agar overlay method, were used. Of special concern were tests of Pseudomonas aeruginosa with moderate susceptibility to carbenicillin; gentamicin, tobramycin, polymyxin B, and colistin were also tested with P. aeruginosa as well as with Klebsiella pneumoniae, Escherichia coli, and Proteus species. The agar overlay method tended to give zones that were somewhat smaller than those given by the Kirby-Bauer method. Both disk methods demonstrated an excellent degree of reproducibility, being at least as reproducible as the standard agar dilution technique for measurement of minimal inhibitory concentrations. Slight changes in carbenicillin disk potency affected the zones of inhibition around P. aeruginosa more than those around susceptible E. coli. This fact suggests that a standard strain of P. aeruginosa would be advantageous for quality control of carbenicillin disk tests.


Differential sensitivity of herpes simplex virus types 1 and 2 to human interferon: antiviral effects of interferon plus 9-beta-D-arabinofuranosyladenine. With use of a standard assay for antiviral compounds, in which the compound to be tested is added after absorption of virus, the minimal inhibitory concentration (MIC) of human interferon for several strain of herpes simplex virus type 2 (HSV-2) is five to 10 times greater than it is for two strains of herpes simplex virus type 1 (HSV-1). This differential susceptibility of HSV types to interferon is found whether tests are done with a liquid overlay and microtiter plates or with agarose overlays and appears to be a distinguishing biological marker. When the MIC of interferon is tested by microtiter or agarose methods and interferon is allowed to incubate for 24 hr before virus is added, values for HSV-1 and HSV-2 are similar and much smaller in magnitude. These results support earlier data indicating that adenine arabinoside and interferon are synergistic against herpes simplex virus type 1 in vitro and indicate that these agents are additive but not synergistic against herpes simplex virus type 2.


Defective initiation of the metabolic stimulation in phagocytizing granulocytes: a new congenital defect. Two patients suffering from recurrent bacterial infections were studied: a boy and a girl from one family, children of apparently healthy parents. The granulocytes of these patients were capable of normal ingestion of latex particles and DNA-anti-DNA immune complexes. When the metabolic changes in these granulocytes during phagocytosis of latex particles were studied, however, no stimulation of oxygen consumption, superoxide production, or hexose monophosphate shunt activity could be observed. Moreover, zymosan particles were not iodinated. These findings are comparable to those found in chronic granulomatous disease. In sharp contrast to the observations in this latter disease, however, a completely normal stimulation of cell metabolism was found after phagocytosis of IgG-coated latex particles or IgG aggregates. Since latex and IgG-coated latex were equally well ingested, this means that the absence of metabolic stimulation after uptake of tatexf metabolic stimulation after uptake of latex must be due to a defect in the triggering of the oxidative burst. As far as we know, this is the first time that a defect in the triggering of the metabolic stimulation during phagocytosis could be demonstrated. Moreover, these finding suggest that adherence and subsequent ingestion of particles are in themselves not sufficient for the metabolic stimulation of granulocytes.


Abnormal pattern of bactericidal activity of neutrophils deficient in granules, myeloperoxidase, and alkaline phosphatase. The morphology, cytochemistry, metabolism, and bactericidal function of neutrophilic polymorphonuclear leukocytes (PMN) from a patient with subacute myelogenous leukemia were evaluated. The patient's mature PMN were deficient in granules and staining reactivity for myeloperoxidase (MPO) and alkaline phosphatase (LAP). These cells killed Staphylococcus aureus in an abnormal pattern when they were challenged with various increasing multiples of bacteria per neutrophil. At a low ratio of challenge (1.25 bacteria per neutrophil) the MPO-LAP-deficient PMN killed only 18 +/- 6 per cent (mean +/- 1 S.D.) (normal, 79 +/- 7) of the initial bacterial inoculum. As the PMN were challenged with higher ratios of bacteria per cell, the bactericidal effectiveness of the hypogranular PMN improved. At a 50:1 ratio the patient's cells killed within the normal range (28 +/- 10 per cent vs. normal of 48 +/- [mean +/- 1 S.D.]). Although rates of glucose oxidation and oxygen consumption by patient or control PMN stimulated with comparable ratios of heat-killed bacteria were the same, only minimal metabolic enhancement was produced in the MPO-LAP-deficient PMN by lower ratios. In contrast, higher ratios produced a marked increase in both of these metabolic activities indicating a major metabolic response to multiple ingestions by the patient's PMN. These observations may reflect the activation of compensatory microbicidal mechanisms available to the MPO-LAP-deficient PMN only when challenged by large multiples of bacteria.


Purification and characterization of Leydig cells from rat testes. An LH-responsive Leydig cell preparation (containing 6+/-2% Leydig cells) was obtained by collagenase treatment of rat testis. Centrifugation of this cell preparation through a 13% Ficoll solution for 10 min at 1500 g resulted in a four times purification of the Leydig cells, with a concomitant increases in steroidogenic activity. Addition of 0-2% albumin to the 13% Ficoll solution, adjusted to 280 mosmol/l, resulted in a further twofold purification of the Leydig cells paralleled by a twofold increase in steroidogenic activity. Centrifugation of these Ficoll-albumin-purified Leydig cells through a 6% dextran solution for 2 min at 100 g resulted in a further 1-7 times purification of the Leydig cells. A combination of the two centrifugation steps resulted in a 12-5 times purification of Leydig cells compared with the original crude cell suspension, while an increase in steroidogenic activity of 22-5 times was obtained. This final cell preparation contained 59 +/- 17% Leydig cells (mean +/- S.D., n = 6). The recovery of Leydig cells was 29%. Collagenase treatment of testes deficient in spermatogenesis resulted in a cell preparation with the same steroidogenic activity as Ficoll-purified cells from normal testes. Centrifugation of these cells through a 13% Ficoll solution gave only a limited increase in the steroidogenic activity. Isopycnic centrifugation of the crude cell preparation on a discontinous Ficoll metrizoate gradient resulted in two discrete peaks of Leydig cells, one peak at a density of 1-039-1-055 g/ml and one at a density of 1-068-1-088 g/ml. Both types of cells produced testosterone. In the presence of LH, cyclic AMP production in both types of Leydig cells increased, but testosterone production was only increased by LH in the "denser" Leydig cells and not in the "light" Leydig cells. No difference in sensitivity to LH could be observed between the Leydig cell preparations of different purity. Using a 60 min pre-incubation period the highest testosterone response was obtained with 100-1000 ng LH/ml. The same maximum testosterone response was obtained with 10-100 ng LH/ml when the pre-incubation period was omitted.


Stimulation of intra-uterine growth in rat by alpha-melanocyte-stimulating hormone. We have studied whether endogenous alpha-MSH has a function in stimulating intra-uterine growth in the rat. The approach used was to determine whether or not this hormone is present during the intra-uterine growth spurt, and if binding of endogenous foetal alpha-MSH by antibodies would inhibit this growth. Antibodies against alpha-MSH or ACTH 1-24, either purified or non-purified, induced immunofluorescence in the intermediate lobe of adult male control rats. Using purified anti-alpha-MSH, fluorescence appeared in the foetal intermediate lobe on day 18 of pregnancy, the day that biologically active MSH was first seen. A negative correlation was observed between the pituitary MSH content and foetal body weight only on day 19 of pregnancy. Injection of purified anti-alpha-MSH induced a drop in foetal body weight, but no effect on placental weight was observed. Purified anti-acth 1-24 had no effect upon body weight but caused an increase in placental weight. These results support our previous findings and indicate that endogenous MSH has a function in the stimulation of foetal growth.


Glycerol release in vitro from adipose tissue of obese (ob/ob) mice treated with thyroid hormones. Glycerol release from adipose tissue of adult genetically obese (ob/ob) mice was greater than that from adipose tissue of lean litter-mates. However, a higher concentration of adrenaline was required to stimulate lipolysis in ob/ob adipose tissue. Since oxygen consumption and the uptake of radioactive iodine by the thyroid were reduced and the biological half-life of thyroidal radio-iodine was lengthened, it is suggested that ob/ob mice are hypothyroid. The poor sensitivity of ob/ob adipose tissue to low doses of adrenaline was improved by treatment with either tri-iodothyronine or thyroxine. The lipolytic responses to both dibutyryl 3':5'-cyclic adenosine monophosphate and aminophylline were enhanced in thyroid hormone-treated lean and ob/ob mice. A higher dose of tri-iodothyronine was required to enhance lipolysis in ob/ob mice than in lean controls. The possibility that hypothyroidism is responsible for the change in response of ob/ob adipose tissue to lipolytic hormones is discussed.


Effect of bilateral adrenalectomy and corticosteroid therapy on the secretion of corticotrophin-releasing factor activity from the hypothalamus of the rat in vitro. The rat hypothalamus in vitro preparation was used to investigate the effect of bilateral adrenalectomy, with and without replacement therapy, on the release of corticotrophin-releasing factor(CRF). Corticotrophin-releasing factor was estimated using 48 h basal hypothalamic lesioned assay rats and corticosterone production of excised adrenals was used as the end point. Bilateral adrenalectomy resulted in depletion of hypothalamic CRF content within the first 2 h after the operation but this effect was prevented by replacement therapy with corticosterone. Thereafter, the hypothalamic CRF content returned to values not significantly different from the intact control level. Bilateral adrenalectomy caused an increase in both basal and acetylcholine-induced release of CRF and it is suggested that corticosteroids exert a negative feedback effect on the hypothalamus.


Pituitary-adrenal function in thyroparathyroidectomized male and female rats. The influence of thyroid hormones on pituitary-adrenal function was assessed by studying several aspects of adrenocortical function approximately 30 days after thyroparathyroidectomy (TPTx). Both male and female rats showed evidence of rhythmic adrenocortical activity; peak plasma corticosterone levels occurred just before the dark phase of the lighting schedule. Only the amplitude of the rhythm appeared altered by TPTx. Peak plasma corticosterone levels in TPTx male and female animals were less (P less than 0.05) than corresponding levels in intact control rats. Both sexes showed significant responses to stress, but the morning stress response in TPTx females was less (P less than 0.01) than the stress response in intact controls. Concomitant with the reduced stress response, the adrenocortical response to exogenous ACTH was reduced in TPTx female rats.


Mouse mammary tumor virus in hybrids from strains C57BL and GR: breeding test of backcross segregants. F1 and F2 and first backcross hybrids and second backcross families of the high mammary tumor incidence strain GR and the low incidence strain C57BL were examined for the segregation of mouse mammary tumor viral (MMTV) expression. Although GR has been reported to transmit MMTV as a single dominant gene, several lines of evidence suggest there are multiple genetic factors that influence MMTV expression, MMTV expression as measured by double antibody radioimmunoassay for MMTV p14 segregated in 106 first backcross progeny at a 60:40 ratio, intermediate between what would be expected for either a single or two gene hypothesis. In female second backcross progeny of either male or female first backcross, a heterogeneous pattern of expression was noted that does not fit any simple Mendalian pattern. From an analysis of serial lactations of first backcross and second backcross families, it appears that all hybrid females contain MMTV proviral information that may be expressed either at late lactations or in a variable proportion of progeny mice. These combined results are most consistent with a vertically transmitted genome regulated by multiple factors in these crosses.


Normal rabbit alveolar macrophages. I. The phagocytosis of tubular myelin. Normal rabbit alveolar macrophages are engorged with large, dense inclusions which contain whorls of myelin figures, suggesting an exogenous source of polar lipids in their diet. One contributory source of such lipids is surfactant, since macrophages were seen ingesting tubular myelin and vacuoles containing remnants of it were found in the cytoplasm. Thus, as indicated previously in kinetic studies, it appears that alveolar macrophages participate in the turnover of surfactant. However, the relative importance of the macrophage in comparison to other pathways of surfactant removal remains to be determined. It is also noteworthy that although tubular myelin and myelin figures were abundant in the fixative used to wash out the lungs, bacteria were not found in it or in the macrophages. Thus, removal of obsolete surfactant may prove to be one of the mojor endocytic functions of alveolar macrophages.


Normal rabbit alveolar macrophages. II. Their primary and secondary lysosomes as revealed by electron microscopy and cytochemistry. In this investigation, vacuoles containing tubular myelin proved to be digestive compartments with cytochemical reactivity for acid phosphatase and arylsulfatase. These cytochemical markers identify the secondary lysosomes, known to contain enzymes capable of hydrolyzing phospholipids like surfactant. Therefore, it appears that alveolar macrophages possess the enzymatic machinery for the degradation of the tubular myelin found in their digestive vacuoles. Although it thus appears evident that alveolar macrophages participate in the turnover of surfactant, the quantitative significance of this route of disposal is undetermined. This investigation has also established that acid hydrolases, so prominently displayed in the secondary lysosomes, are also localized in the rough endoplasmic reticulum and in Golgi-endoplasmic reticulum-lysosomes (GERL). Moreover, small vesicles which are produced from GERL serve as primary lysosomes in transporting digestive enzymes to the vacuoles.


The effects of cytochalasin and colchicine on interferon production. The present study was undertaken to investigate the mechanism of interferon production by mouse spleen cells co-cultivated with BHK-HVJ cells, i.e. baby hamster kidney cells persistently infected with the HVJ (or Sendai) strain of para-influenza I virus. Cytochalasin B appears to inhibit an early stage in the process and colchicine a relatively late stage. It is suggested that microfilaments and microtubules may play in important role at an initial stage of interferon production in this system.


Synthesis of Sendai virus polypeptides by a cell-free extract from wheat germ. The '18S' RNA population from Sendai virus-infected cells efficiently directed the synthesis of three virus-specific polypeptides P, NP and M, in wheat germ cell-free extracts. In agreement with previous results obtained with a reticulocyte extract, there was little or no production of virus glycopolypeptides. Analyses of tryptic peptides revealed close correspondence between the primary structures of NP and M made in vitro and the authentic virus polypeptides.


A partial denaturation map of herpes simplex virus type 1 DNA: evidence for inversions of the unique DNA regions. Partial denaturation maps of 30 HSV-I DNA molecules have been obtained using a procedure designed to avoid possible hydrolysis of the DNA at alkalilabile bonds. From the denaturation pattern of the long unique DNA region these molecules were divided into two groups comprised of 16 and 14 molecules. Histogram plots relating the precentage denaturation to position on the DNA for these two groups were aligned in a manner appropriate to the HSV-I genome model. It was apparent that these groups had the orientation of the long region inverted with respect to each other. Similarly, from the denaturation maps of the short unique region, the molecules were divided into two groups each comprising 15 molecules. Alignment of the histogram plots of these groups indicated that the orientation of the short region was inverted in one group relative to the other. These partial denaturation data confirm the presence of four HSV-I genome arrangements resulting from the possible combinations of inversions of the two unique DNA regions.


Plaque formation with influenza viruses in dog kidney cells. Infectious influenza and parainfluenza viruses were produced as plaql passages of egg-grown viruses in MDCK monolayers. Virus titres of 10(6) to 10(11) p.f.u./ml were obtained for several A and B strains of influenza and parainfluenza viruses.


Characterization of pig rotavirus RNA.

Characterization of pig rotavirus RNA. Pig rotavirus was purified from faeces. The RNA from this virus was resistant to pancreatic ribonuclease, indicating that it is double-stranded. When electrophoresed on polyacrylamide-agarose gels, pig rotavirus RNA migrated as 9 bands comprised of 11 or 12 RNA segments with a total mol. wt. of approx. 11 X 10(6). Co-electrophoresis experiments revealed that the RNAs from the pig virus and two isolates of the calf rotavirus were indistinguishable.


Virus-like particles in bovine sera for tissue culture. Virus-like particles were found in nine different bovine sera for tissue culture from commercial suppliers. These particles were spherical with an overall diam. between 70 and 95 nm. After negative staining, surface projections of about 11 to 12 nm were clearly seen. One of the nine sera was positive in an Ouchterlony test with antisera against bovine viral diarrhoea virus.


Concanavalin A agglutinability of a temperature-sensitive mutant of Rous sarcoma virus. Concanavalin A (Con A) concentration dependence of agglutinability of a mutant of Rous sarcoma virus which is temperature-sensitive for transformation was examined. Con A agglutinability of the virion was quantitatively expressed by measuring radioactivities of 60 to 70 S RNA extracted from Con A-agglutinated material. The mutant grown at a permissive temperature (35 degrees C) agglutinated at a significantly lower concentration of Con A, compared with that at a non-permissive temperature (40 degrees C).


Analysis of the RNA species isolated from defective particles of vesicular stomatitis virus. Serial high multiplicity passage of a cloned stock of vesicular stomatitis virus was found to generate defective interfering particles containing three size classes of RNA, with sedimentaiton coefficients of 31 S, 23 S and 19 S. The 31 S and 23 S RNA species were found to be complementary to both the 12 to 18 S and 31 S size classes of VSV mRNAs. The 19 S class of RNA was found to be partially base-paired. All three RNA species were found to contain ppAp at their 5' termini.


Effects of interferon on cell and virus growth in transformed human cell lines. The anticellular and antiviral effects of human leukocyte interferons were studied in vitro in the transformed human embryonic cell lines. RSa and RSb. The growth of these cells was inhibited and they began to deteriorate about 48 h after treatment with 500 units/ml of interferon. When interferon was washed out within 48 h, their growth recovered gradually. The effects of interferon on cell growth depended on the amount of interferon added per cell. A subline, named IFr, was isolated which grows in the presence of 2000 units/ml of interferon, whereas growth of vesicular stomatitis virus in these cells is suppressed by 10 units/ml of interferon, just as in the parent cells. The anticellular and antiviral effects of interferon on IFr cells are discussed in relation to cell surface receptors.


Factors involved in the expression of cowpox virus-specific antigen in Sendai virus carrier cells. Formation of cowpox virus-specific cell surface antigen (CPVS-ag) HVJ (Sendai virus) carrier cells compared to that in parent cells. Temperature shifts from 32 to 35 to 37 degrees C for these carrier cultures reduced this enhancing activity, making them equivalent to parent cells. The eclips phase and one-step growth of CPV in the carrier cells were shorter than in normal cells. Extracts of carrier cells exhibited a stimulating activity causing the temporary rise and subsequent lowering of CPV infectivity in their reactions with CPV in vitro, suggesting a cellular acceleration of CPV uncoating. The S-ag forming ability of these carrier cells did not decrease as much as that of parent cellsin the presence of actinomycin D or puromycin. The results indicate that persistent infection with HVJ, especially the temperature-sensitive variant, promotes the first step of intracellular growth of CPV, resulting in enhanced formation of S-ag.


The abdominal ganglion of Aplysia brasiliana: a comparative morphological and electrophysiological study, with notes on A. dactylomela. The ultrastructure and electrophysiological properties of neurons in the abdominal (visceral) ganglion of the marine opisthobranch gastropod Aplysia brasiliana have been investigated to determine whether this preparation compares favorably with the well studied A. californica for neurobiological research. In general, the topography, morphology and physiological characteristics, including synaptic connections, of neurons in this ganglion are quite similar to those of A. californica. There is close correspondence between the two animals in terms of each of the identified cells or neuronal clusters in the ganglion, including the presence of the cell L10 (interneuron I) in A. brasiliana which makes synaptic connections comparable with those in A. californica. New follower cells of this interneuron have been found in A. brasiliana. This species offers some advantages in that the connective tissue surrounding the ganglion is thinner and more transparent, making cell identification and penetration easier. A. brasiliana appears to exhibit the behaviors of A. californica that have been used in previous functional analyses of neural circuits. In addition, this species swims and exhibits a "burrowing" activity less commonly seen in A. californica. The rich repertoire of behaviors and accessibility of large identifiable and functionally interconnected neurons makes this species of Aplysia an excellent model preparation for future neurobiological studies. Similar, less thorough, investigations of the abdominal ganglion of A. dactylomela indicate that this species is also very similar to A. californica in terms of the identified cells in the abdominal ganglion.


The identification of two inhibitory cells in each segmental ganglion of the leech and studies on the ionic mechanism of the inhibitory junctional potentials produced by these cells. The present study identifies a pair of inhibitory cells that are located on each anterolateral margin of a leech segmental ganglion. These cells, which we label as cells 119, are electrically interconnected. These cells give rise to inhibitory junctional potentials (ijp's) in contralteral longitudinal body wall muscle cells. The latencies of the ijp's following spikes in cell 119 are variable. The ijp's are caused by transient increases in premeability to the Cl- ion. Previous studies demonstrated that 5-HT causes a hyperpolarization of body wall muscle cells by increasing the permeability of muscle membrane to the Cl- ion. Accordingly, 5-HT was searched for in the 119 cell bodies, but autoradiography, fine structure, and gas chromatography-mass spectrometry gave no indication that 5-HT was present in these cells. However, the variable latencies of the ijp's may indicate that there is a neuron interposed between cell 119 and the muscle cells. If this is the case, then the interposed neuron should be analyzed for 5-HT. Further experiments to locate the terminals of cells 119 and the cell bodies of the presumed interposed neurons are thus desirable.


Demyelinative myelopathy in mice induced by the DA virus. An attenuated tissue culture adapted strain of DA virus, an agent related to the Theiler murine encephalomyelitis viruses (TMEV), was used to induce a chronic myelopathy in mice. Spastic paraparesis first appeared 5 months after weanings were inoculated intracerebrally with the virus. None died as a direct result of the infection, and none improved once paretic. The major pathological change in these mice was demyelination of thoracic segments of spinal cord. No clinical illness or demyelinative pathology were detected during the first 4 months after inoculation. Encephalitic virus was present in brain and spinal cord as late as 10 months after inoculation. No neutralization antibody activity to DA virus was present in sera from 10 patients with amyotrophic lateral sclerosis, 10 with multiple sclerosis, or in 10 controls.


Lipid abnormalities in hereditary neuropathy. Part I. Serum non-polar lipids. The non-polar lipids from sera of 54 patients, with various types of hereditary motor and sensory neuropathies, and from 72 healthy subjects were evaluated. A small but highly significant decrease in the percentage of linoleate to total fatty acids in both cholesteryl ester and triglyceride fractions was found in the sera of the neuropathy patients, except in those who had dominantly inherited sensory neuropathy (HSN-I) and who had spinocerebellar degeneration with retinitis pigmentosa and other features (SpC+). A significant decrease of serum lecithin-cholesterol acyltransferase activity was also found in those patients with hereditary motor and sensory neuropathies, Type I and Type II (two types of peroneal muscular atrophy). The biochemical basis of these abnormalities is not apparent. The biochemical abnormalities reported here have been found in several neurologic disorders and hence are unlikely to be disease-specific.


Spinal changes in the neuropathy of thallium poisoning. A case with neuropathological studies. Thallium poisoning in a man of 32 years resulted in typical peripheral neuropathy, skin rash and alopecia. Post-mortem studies showed widespread loss of peripheral nerve fibres and chromatolysis in spinal neurones. The distal nature of the nerve fibre degeneration in thallium poisoning and the sensitivity of long fibres is emphasized in this case.


Neurochemical findings in adreno-leukodystrophy. The results of neurochemical examination of brain tissue derived from 2 cases of adreno-leukodystrophy have been presented. Both white and grey matter contained suadanophilic material. Although free fatty acid was also present, cholesteryl ester accounted for the bulk of the sudanophilic material. Total cholesterol, galactolipid and phospholipid content was reduced in both white and grey matter. The lipid loss was particularly severe from white matter indicative of considerable demyelination. Cholesterol was found to be the only major sterol present in white or grey matter. Subcellular fractionation of the diseased white matter resulted in myelin and two related fractions, one of which was very fatty and was rich in steryl ester. Morphological examination of myelin indicated loosely-packed lamellae. All of 3 fractions had adenosine 2',6'-cyclic nucleotide-3'-phosphohydrolase activity. Myein and the fraction not rich in cholesteryl ester had discernable basic protein bands when examined by polyacrylamide gel electrophoresis. Analysis of the fatty acid composition of choline and ethanolamine glycerophospholipids indicated a general increase of saturated fatty acids, relative to control values and a decrease in long-chain fatty acids. Examination of sphingomyelin fatty acids also demonstrated a loss of long-chain fatty acids. The fatty acid composition of the cholesteryl esters from white and grey matter differed. The findings indicate generalized damage to the brain, both of white and grey matter, with the damage to the white matter being much more severe. No abnormal sterol or other lipid was isolated.


Concentric laminated bodies. Ultrastructural demonstration of muscle fiber type specificity. Concentric laminated bodies were identified in the skeletal muscle of 3 children affected by muscle weakness and hypotonia with probable cerebral involvement. The mean Z-line and M-line widths from each of 11 muscle fibers containing the concentric laminated bodies were calculated. The mean Z-line widths were 61-81 nm and the mean M-line widths from 7 out of 11 fibers were 54-65 nm. According to our present system of fiber typing, the majority of these fibers would be classified as subtype IIB. The possible pathogenesis and fiber type specificity of the concentric laminated body is discussed.


Peripheral neuropathy complicating primary hyperoxaluria. A patient with chronic renal disease due to primary hyperoxaluria developed a rapidly progressing motor neuropathy with marked impairment of nerve conduction. Pathological studies demonstrated the presence of both axonal degeneration and segmental demyelination, together with the presence of oxalate crystals within axons. It is suggested that the development of peripheral neuropathy complicating hyperoxaluria is a consequence of the increased life-span mad possible by haemodialysis.


Recessive hereditary sensory neuropathy. Clinical features in 2 cases of a recessive form of hereditary sensory neuropathy and the light and electron microscopy of sural nerve biopsy in 1 of them are described. The patients showed symptoms typical of this form of the disease; it should be stressed however that the loss of cutaneous sensation appeared to be limited to the distal parts of the lower extremities and involved all modalities of cutaneous sensation. Histological examination of sural nerve revealed a marked reduction in the number of myelinated fibres due to Wallerian-like axonal degeneration, of which various stages were represented. In addition, segmental demyelination, probably secondary to axonal changes, was seen. The unmyelinated fibres were also involved but to a lesser degree than the myelinated fibres. The observations indicate a progressive nature of the pathological process.


Alteration of the maximal activity of the gluconeogenetic enzyme fructose-1,6-diphosphatase of skeletal muscle by cross-reinnervation. A histochemical and biochemical investigation of fatiguability-related aspects. Fatiguability is a determining characteristic of different muscle fibre types. An important aspect, indirectly related to fatiguability, gluconeogenesis, was investigated by observing fructose-1,6-diphosphatase (FDP) activity in experimental models prepared in rabbits by cross-reinnervation of the fatigue-resistant m.soleus (postural muscle) and the highly fatiguable m.flexor digitorum longus (fast muscle -- m.fdl). The resultant reprogramming of the m. soleus was associated with greatly intensified FDP activity. Changes in the m.fdl 6-9 months after cross-reinnervation indicated a shift in the opposite direction. The study adds some data on the much neglected state of fatiguability to the otherwise much explored field of alien reinnervation.


[Recurrent peripheral nerve palsies and concomitant chronic neuropathy. Neuromusclar electrophysiologic study and neuropathologic examination: apropos of 5 cases]. The authors report 5 personal cases (3 in the same family) of patients suffering from recurrent peripheral nerve palsies usually induced by pressure and review the relevant literature. There is evidence in such cases of a mild, chronic, diffuse, subclinical neuropathy which can be recognized by electrophysiological examination. Examination of peripheral nerve biopsy material in such cases shows evidence of both axonal and Schwann cell abnormalities with evidence of neurogenic atrophy of skeletal muscles. This chronic subclinical neuropathy which is often familial can be distinguished from other chronic primary neuropathies with or without myelin sheath proliferation and hypertrophy.


Cerebellar glioblastoma in childhood. Case report. Ten childhood cerebellar glioblastomas have been reported previously in the world literature. The authors report a case arising de novo in a 9-year-old child and stress the rarity of this tumor.


myo-Inositol metabolism in the neonatal and developing rat fed a myo-inositol-free diet. Neonatal rats of the Holtzman strain, 6 days of age, were fed a myo-inositol restricted liquid formula by gastric intubation for 10 days, after which they were fed a purified myo-inositol-free diet until they were 72 days old. No differences in weight gain were observed between myo-inositol/100 ml of formula or 150 mg myo-inositol/100 g diet. Most tissues examined from rats fed the myo-inositol deprived formula and diet had lower free myo-inositol levels than the controls with the exception of the liver. Despite reduced free and lipid-bound myo-inositol in the liver, there was no evidence of fatty liver in the young rats at any age. The cerebrum and cerebellum of myo-inositol deprived rats had normal myelination and mitochondriogenesis as judged by the levels of 2',3'-cyclic nucleotide-3'-phosphohydrolase (EC 3.1.4.1) and fumarase (EC 4.2.1.2) activity, respectively.


Neurochemical changes in Leigh's disease. A series of children with Leigh's disease had normal hepatic pyruvate carboxylase activity, increased cerebral thiamine diphosphate, and decreased cerebral thiamine triphosphate. These thiamine esters were normal in liver. The author suggests that the histologic changes of Leigh's disease, as well as the similar changes of Wernicke's disease, could be due to a deficiency of cerebral thiamine triphosphate.


Cytochemical localization of peroxidase activity in the mitochondria of Hymenolepis diminuta. A cytochemical study of mitochondria of Hymenolepis diminuta indicates the presence of a mitochondrial peroxidase. Utilizing a 3,3'-diaminobenzidine (DAB) medium at pH 9.7, the reaction product is localized in the intracristal space, and between the inner and outer membranes of the mitochondria. No inhibitory effects are exerted on the peroxidase reaction by cyanide, azide, or aminotriazole. In addition, the mitochondria appear to have an enzyme which is cytochemically similar to vertebrate cytochrome c-oxidase. The possible physiological significance of the peroxidase is discussed.


Oxidation of selected pteridine derivatives by mamalian liver xanthine oxidase and aldehyde oxidase. Considerable information is available concerning the oxidation of pteridine derivatives by bovine milk xanthine oxidase, but few investigations have been carried out on the oxidation of such compounds by mammalian liver xanthine oxidase and the related aldehyde oxidase. Xanthine oxidase, obtained from rat liver, oxidizes a variety of substituted amino- and hydroxypteridines in a manner identical to that previously observed for milk xanthine oxidase. For example, 2-aminopteridine and its 4- and 7-hydroxy derivatives were oxidized efficiently to 2-amino-4,7-dihydroxypteridine (isoxanthopterin) by the rat liver enzyme, and 4-aminopteridine and its 2- and 7-hydroxy derivatives were oxidized to 4-amino-2,7-dihydroxypteridine.4-Hydroxypteridine and the isomeric 2- and 7-hydroxypteridines were oxidized by rat liver xanthine oxidase to 2,4,7-trihydroxypteridine. Rabbit liver aldehyde oxidase, but not rat liver xanthine oxidase, was able to catalyze the oxidation in position 7 of 2,4-diaminopteridine and its 6-methyl and 6-hydroxymethyl derivatives. 2-Aminopteridine and 4-aminopteridine were both oxidized to the corresponding 7-hydroxy derivatives in the aldehyde oxidase system; 2-amino-4-hydroxypteridine appeared to be a minor product in the oxidation of 2-aminopteridine by rabbit liver aldehyde oxidase. Both aldehyde oxidase and xanthine oxidase were able to catalyze the oxidation of 2-amino-6,7-disubstituted pteridines to the corresponding 4-hydroxy derivatives; 4-hydroxy-6,7-disubstituted pteridines were oxidized in position 2 by both enzymes. 4-Amino-6,7-disubstituted pteridines were not oxidized by either enzyme. 2-Amino-4-methylpteridine was oxidized in position 7 by aldehyde oxidase but was not an effective substrate for xanthine oxidase; 2-hydroxypteridine and 7-hydroxypteridine were not oxidized to a detectably extent by aldehyde oxidase. All oxidations mediated by xanthine oxidase were strongly inhibited by allopurinol (4-hydroxypyrazolo[3,4-d]pyrimidine), and all oxidations mediated by aldehyde oxidase were inhibited by menadione (2-methyl-1,4-naphthoquinone). Rat liver xanthine oxidase and, to a lesser extent, rabbit liver aldehyde oxidase were inhibited by 4-chloro-6,7-dimethylpteridine; 2-amino-3-pyrazinecarboxylic acid inhibited xanthine oxidase but not aldehyde oxidase. The oxidations of 2- and 4-aminopteridines by aldehyde oxidase resulted in concomitant reduction of cytochrome c.


Isoproterenol-induced relaxation, phosphorylase activation and cylic adenosine monophosphate levels in the polarized and depolarized rat uterus. There is some evidence in the literature that catecholamines relax uterine and other types of smooth muscle by increasing tissue levels of cyclic adenosine monophosphate (cyclic AMP). In the present study, isoproterenol completely relaxed uterine strips obtained from estrogen-primed rats and also increased tissue levels of cyclic AMP and phosphorylase a. In uterine strips depolarized and put into contracture for 15 minutes by 127 mM K+, isoproterenol did not increase phosphorylase a or cyclic AMP but was still capable of producing relaxation. When uterine strips were exposed to the methoxy derivative of verapamil, D-600, a compound known to prevent the influx of calcium, the uterus relaxed completely without an increase in cyclic AMP. The addition of isoproterenol at this stage resulted in an increase in cyclic AMP similar to that noted in nondepolarized uterine strips. The addition of 127 mM K+ also resulted in time-dependent biochemical changes as well as contracture. Cyclic AMP was increased 3-fold after 2 minutes of K+ depolarization and phosphorylase a was increased as well. The increase in cyclic AMP was prevented by propranolol but propranolol did not affect the contracture response to K+. D-600 prevented contracture but did not affect the K+-induced increase in cyclic AMP. The data suggest that an increase in whole tissue levels of cyclic AMP are not necessary in order for isoproterenol to relax depolarized rat uterine strips. The data also suggest that intracellular calcium levels can affect the level of cyclic AMP in the rat uterus.


Regulation in the central norepinephrine neurotransmission induced in vivo by alpha adrenoceptor active drugs. The level of the two major norepinephrine metabolites, 3-methoxy-4-hydroxyphenylglycol (MOPEG) and 3,4-dihydroxyphenylglycol (DOPEG), was estimated in the central nervous system of rats to study receptor-mediated regulation of release in vivo as reflected in biochemical changes. The norepinephrine receptor stimulating drug clonidine (0.02-0.5 mg/kg) decreased the level of endogenous total MOPEG. The accumulation of 3H-MOPEG and 3H-DOPEG was decreased by clonidine (0.5 mg/kg) regardless of whether 3H-tyrosine or 3H-dopamine was used as precursor of 3H-norepinephrine. In contrast to clonidine, the two alpha adrenoceptor blocking drugs, phenoxybenzamine (20 mg/kg) and aceperone (20 mg/kg), induced an increase in endogenous total MOPEG and also an increase in 3H-MOPEG and 3H-DOPEG regardless of the precursor used. These results indicate that clonidine decreases the release of norepinephrine in vivo and that phenoxybenzamine and aceperone increase the release of norepinephrine. Clonidine inhibited completely the effect of phenoxy benzamine or aceperone on endogenous MOPEG. On the contrary, it was not possible to block completely the effect of small doses of clonidine by pretreatment with either phenoxybenzamine, yohimbine (2 mg/kg) or a high dose of aceperone. These results indicate that clonidine may act on a different target than the alpha adrenoceptor blocking drugs. In vitro experiments with occipital cortex synaptosomes did not indicate a direct effect of clonidine on tyrosine hydroxylation in noradrenergic nerve terminals.


Studies on the comparative pharmacology and selective toxicity of tricaine methanesulfonate: metabolism as a basis of the selective toxicity in poikilotherms. Tricaine methanesulfonate, administered at a dose of 150 mg/kg i.p., produced a flaccid paralysis and loss of the righting reflex in a number of poikilothermic species including the frog. Leopard frogs (Rana pipiens) given 150 mg/kg i.p. regained the righting reflex at 113 +/- 28 (S.D.) minutes after injection. A similar dose administered i.p. to mice produced no apparent pharmacological response. The biological half-life (T1/2) of tricaine in frogs (R. pipiens) was about 70 minutes at temperatures of 23 and 37.5 degrees C; at 7 degrees C the T1/2 was 309 minutes. In contrast, in the mouse the drug was metabolized so rapidly that 5 minutes after i.p. administration of 5 mg of tricaine methanesulfonate (250 mg/kg) none of the unchanged drug could be recovered from the animal. It was, however, recovered quantitatively as Bratton-Marshall reacting metabolites, including m-aminobenzoic acid. Incubations of tricaine with serum from bullfrogs, mice and humans indicated that the drug was metabolized to m-aminobenzoic acid with an apparent Km of 3 X 10(-3) M for the reaction. The Vmax for incubations of the drug with bullfrog and human serum was 40 nmol/min/ml of serum, whereas in mouse serum it was 93 nmol/min/ml of serum. In vitro studies with liver homogenates showed that mouse liver metabolized tricaine 39 times more rapidly than frog liver. We conclude, therefore, that the liver is the major site of tricaine hydrolysis in mammals and that the selective toxicity of tricaine for poikilotherms is a consequence of their slower rate of hepatic biotransformation of tricaine.


Studies on the narcotic receptor in the guinea-pig ileum. Studies were conducted on the development and loss of tolerance to morphine in the coaxially stimulated guinea-pig ileum. In ilea from guinea pigs made tolerant to morphine by the procedure of morphine-pellet implantation, the morphine-naloxone pA2 was decreased from 8.5 to 7.6, suggesting a qualitative rather than a quantitative change in the receptors. This change in the pA2 was in the opposite direction from that previously observed with analgesic receptors. Three hours after the administration of a single injection of morphine to the guinea pig, the ileum showed tolerance to morphine, which disappeared by 6 hours. With naloxone as the antagonist, the narcotic analgesics, morphine, methadone, etorphine and levorphanol, yielded higher pA2 values than the narcotic antagonist analgesics, nalorphine, pentazocine and cyclazocine, a result similar to that seen with the analgesic receptors. However, the interaction between naloxone and the narcotic antagonists in the ileum differed from that in the central nervous system when the slopes of the pAx plots were examined. Thus, although the interaction of analgesics with the ileal receptors appears to correlate with the acute effects of the drugs, caution must be exercised to use the ileal receptors as models of analgesic receptors for the study of chronic narcotic effects, i.e., tolerance and dependence.


The effect of cyclic nucleotides on the release of 3H-dopamine from rat striatal slices. Slices (1.0 mm thick and 0.4 cm in diameter) obtained from rat neostriatum were preincubated with 3H-dopamine (3H-DA; 0.8 mugM) for 30 minutes and then superfused at a rate of 1.0 ml/min in glass chambers (volume, 2 ml) for varying lenghts of time prior to  electrical stimulation. Superfusate effluents were continuously collected and analyzed for 3H-DA and 3H-metabolites by liquid scintillation spectrometry after separation by alumina and Dowex 50 column chromatography. 3H-DA represented only a small proportion of the spontaneous overflow of radioactivity (9.8%) but represented the largest portion recovered in the tissue (55%). After electrical stimulation of the slices (biphasic square-wave pulses, 50 pulses/sec; 1.1-msec duration; supramaximal voltage), there was a marked increase in 3H-DA and all 3H-fractions. The percent increase of 3H-DA, 3H-O-methylated + 3H-O-methylated deaminated, 3H-dihydroxyphenylacetic acid and 3H-dihydroxylphenylethanol was 549, 232, 215 AND 246%, respectively. The addition to the superfusion medium of dibutyryl adenosine 3:5-monophosphate (DBcAMP) in the presence or absence of phosphodiesterase inhibitors (3-isobutyl-1-methylxanthine, 0.5 mM, or aminophylline, 5 mM) and cyclic adenosine monophosphoric acid in the presence but not the absence of the phosphodiesterase inhibitors potentiated the electrically induced release of 3H-DA. Neither cyclic nucleotide had any effect on the spontaneous overflow of 3H-DA or metabolites, the metabolism of 3H-DA or the uptake of 3H-DA into dopaminergic neurons. Guano sine 3:5-monophosphate was without effect on the electrically induced release of 3H-DA. The ED50 for the DBcAMP potentiation of hte electrically induced release of 3H-DA was 6 mugM. These results extend to the central nervous system the possibility that adenosine 3-5-monophosphate may play a role in the regulation or modulation of monoaminergic synaptic transmission.


Adenosine inhibition of isolated rabbit ileum and antagonism by theophylline. The spontaneously contracting isolated rabbit ileum was used to study adenosine stimulated receptors. The inhibitory effects of adenosine were not reduced by pretreating the rabbits with either reserpine of 6-hydroxydopamine which were used to eliminate adrenergic function. Similarly the addition of tetrodotoxin to the muscle bath had no effect on the ability of adenosine to produce its inhibitory response. Of the compounds tested for agonistic activity, adenosine and ATP were the most potent (ED50  6 X 10(-7) M). The inhibition by adenosine was antagonized by both theophylline and caffeine in a surmountable manner. Theophylline analogs with charged substituents in position 7 were without antagonist activity. The results suggest that 1) receptors for adenosine or adenine nucleotides are located on the smooth muscle cells of rabbit ileum, 2) receptor stimulation requires and intact adenosine moiety and 3) methylxanthines exert their antagonistic effects by acting as competitive antagonists.


Destruction of sympathetic nerve terminals by 6-hydroxydopamine: protection by 1-phenyl-3-(2-thiazolyl)-2-thiourea, diethyldithiocarbamate, methimazole, cysteamine, ethanol and n-butanol. 1-Phenyl-3-(2-thiazolyl)-2-thiourea (PTTU) administered i.p. to mice prevented the neurodegenerative actions of subsequently injected (1 hour later) 6-hydroxydopamine (6-OHDA) or 6-aminodopamine on peripheral adrenergic nerve terminals. Destruction of nerve terminals was studied in vitro in the left atrium by measuring the accumulation of 3H-norepinephrine (3H-HE), and in the iris by both 3H-NE accumulation and fluorescence microscopy methods. Strong protection was observed at 4, 24 and 72 hours after 6-OHDA. The degree of protection was dose-dependent and showed step-wise decrements for concentrations of PTTU below 200 mg/kg (viz., 100, 50 and 20 mg/kg) or concentrations of 6-OHDA-HBr above 7.5 mg/kg (viz., 10, 20 and 50 mg/kg). Protection also fell off at time intervals greater than 1 hour after administration of PTTU (viz., 3 and 5 hours). The appearance (fluorescence microscopy) of the nerve plexus of fully protected mice and the remaining plexus in partially protected mice was essentially normal at 24 hours, except for infrequent large swellings. PTTU proved to be a very effective scavenger of hydroxyl radicals; the formation and scavenging of these radicals was studied by gas chromatography in a system in which the hydroxyl radicals (which were generated during the autoxidation of 6-aminodopamine) gave rise to ethylene, a hydrocarbon gas. Other hydroxyl radical scavengers, namely diethyldithiocarbamate and methimazole, exhibited a protective action on sympathetic nerves in the left atrium; PTTU, diethyldithiocarbamate and methimazole are also recognized as copper chelating compounds. Ethanol, n-butanol and cysteamine, which are well known hydroxyl radical scavengers, also exhibited some degree of protection against 6-OHDA. Blockade of transport of 6-OHDA into sympathetic nerves was ruled out as a protective mechanism by the observation that none of the protective compounds inhibited the accumulation of tritium by the left atrium when 3H-NE was injected in place of 6-OHDA. The mechanism of action for these protective agents has not been definitively established, but scavenging of cytotoxic hydroxyl radicals within neurons may play a significant role.


Effect of Schistosoma mansoni infection on the hepatic drug-metabolizing capacity of mice. Since infections with Schistosoma mansoni cause marked histopathological changes in the liver of the host, the effect of this infection on the hepatic drug-metabolizing function was investigated. Severity of Schistosomiasis was determined by worm counts, duration of infection, egg counts and liver weight increases. To overcome difficulties in homogenizing the livers of infected animals, preincubation of the squashed tissues with collagenase and hyaluronidase was used to prepare homogenates. Key component enzyme activities of the hepatic microsomal drug-metabolizing enzyme system (NADPH-cytochrome c reductase and cytochrome P-450) as well as the representative drug-metabolism activities (aminopyrine N-demethylase, aniline hydroxylase, and benzpyrene hydroxylase) were measured for the whole liver and found to be markedly reduced. However, the measurement of microsomal marker enzyme activities (cytochrome b5 and glucose-6-phosphatase) showed significant elevation. To obtain more precise information about the effect of the schistosome infection on the hepatic drug-metabolizing enzyme system, the total activities of microsomal drug-metabolizing enzymes were related to the total microsomal marker enzyme activities in the homogenate.


Stimulus-secretion coupling: role of cyclic AMP, cyclic GMP and calcium in mediating enzyme (kallikrein) secretion in the submandibular gland. 1. The role of adenosine 3':5'-phosphate (cyclic AMP) and guanosine 3':5'-phosphate (cyclic GMP) as second messengers for the enzyme secretory response evoked by the autonomic neurotransmitters, noradrenaline and acetylcholine, is examined in this in vitro study on the guinea-pig submandibular gland. 2. Noradrenaline increased enzyme (kallikrein) secretion. The initial stimulation of enzyme release appeared to be dose-dependent. The time course of cumulative kallikrein secretion revealed a complex pattern. Isoprenaline and phenylephrine were almost as potent as noradrenaline in releasing kallikrein. Both propranolol and phentolamine were required to fully inhibit the noradrenaline-stimulated enzyme secretion. 3. The cumulative secretion of kallikrein evoked by acetylcholine was dose-dependent. The onset of secretion showed a significantly greater time-lag than that observed with noradrenaline. Atropine effectively blocked the release of kallikrein by acetylcholine. 4. Dibutyryl cyclic AMP stimulated enzyme secretion. Dibutyryl cyclic GMP caused an initial increase which was not maintained. 5. The cyclic nucleotide phosphodiesterase inhibitors, theophylline and papaverine, increased basal kallikrein secretion. The action of the cyclic phosphodiesterase inhibitors on the secretory response to noradrenaline, acetylcholine, dibutyryl cyclic AMP and dibutyryl cyclic GMP was complex. In general, the increase in enzyme release produced by the secretagogues was additively enhanced by both inhibitors. 6. Omission of calcium inhibited both acetylcholine and dibutyryl cyclic GMP stimulated kallikrein release, but to a lesser degree than that of noradrenaline and dibutyryl cyclic AMP. High concentrations of extracellular calcium (10 mM) appeared to enhance the action of acetylcholine. 7. Noradrenaline produced a rise in the intracellular level of cyclic AMP. The increase preceded the stimulated secretion of kallikrein. Of the various adrenergic agonists, noradrenaline and isoprenaline were the most potent, whereas phenylephrine was significantly less effective in raising basal cyclic AMP values. Acetylcholine was without effect, even in the presence of a cyclic phosphodiesterase inhibitor. 8. Acetylcholine and noradrenaline raised intracellular levels of cyclic GMP only when the tissue incubations were performed in the presence of a cyclic phosphodiesterase inhibitor. The increase in cyclic GMP produced by acetylcholine preceded enzyme secretion. 9. Morphological data substantiated the finding that the in vitro release of kallikrein evoked by the secretagogues was associated with the depletion of secretory granules and vacuolations in acinar cells of the gland slices. 10. The molecular mechanisms which control enzyme secretion in the exocrine submandibular gland are discussed. Models are presented for the role of transmitter-specific cyclic nucleotides and calcium in stimulus-secretion coupling.


Endogenous prostaglandins, adenosine 3':5'-monophosphate and sodium transport across isolated frog skin. 1. Sodium transport across isolated frog skin, as measured by the short-circuit current, was decreased by acetylsalicylic acid, mefenamic acid, paracetamol and phenylbutazone. Indomethacin (6 X 10(-6) M) had a biphasic effect on the short-circuit current: a transient increase followed by a sustained decrease. 2. The release of prostaglandin-like material from the skin was reduced by acetylsalicylic acid and indomethacin. Paracetamol caused a significant reduction in the short-circuit current response of the skin to low doses of arachidonic acid, but the response to the highest dose tested was not significantly altered. 3. Indomethacin (6 X 10(-6) M) increased the sensitivity of the skin to applied prostaglandin E1. The other prostaglandin synthetase inhibitors did not have this effect. Indomethacin (6 X 10(-6) M) also enhanced the effect of antidiuretic hormone on the short-circuit current. 4. Indomethacin (30 X 10(-6) M) increased the short-circuit current and diminished the response to applied prostaglandin E1. 5. In sulphate Ringer, theophylline increased the short-circuit current and diminished the response to prostaglandin E1. 6. Prostaglandin E1 increased the levels of cyclic AMP in frog skin and these increases preceded the increases in short-circuit current. There was a seasonal variation in the level of cyclic AMP in the skin: the levels in winter exceeded those in summer. There was also a seasonal variation in the cyclic AMP response to prostaglandin E1: the winter response was greater than that in summer. 7. Indomethacin (6 X 10(-6) M) had a biphasic effect on cyclic AMP levels in the skin, an initial increase followed by a decrease. Indomethacin also potentiated prostaglandin E1 stimulated cyclic AMP accumulation. 8. Theophylline increased cyclic AMP levels in the skin and potentiated prostaglandin E1 stimulated cyclic AMP accumulation. 9. Pre-treatment of the skin with theophylline reversed the effects of cyclic AMP on the short-circuit current and open-circuit potential. 10. It is concluded that endogenous prostaglandins help to maintain sodium transport across isolated frog skin and that the effects of E-type prostaglandins on the short-circuit current are mediated by increased cyclic AMP levels. The transient increase in short-circuit current and the increased skin sensitivity caused by indomethacin (6 X 10(-6) M) are attributed to inhibition of phosphodiesterase activity. The failure of theophylline to potentiate the short-circuit current response of the skin to prostaglandin E1 is attributed to alteration of cyclic AMP action on the skin by theophylline.


alpha-Adrenergic, beta-adrenergic and cholinergic mechanisms for amylase secretion by rat parotid gland in vitro. 1. Rat parotid gland slices, incubated in a balanced, buffered salt solution, were found to be physiologically stable for up to 2 hr with respect to O2 consumption, water content, extracellular space and cation content. 2. The slices could be stimulated to secrete amylase by activation of alpha-adrenergic, beta-adrenergic or muscarinic cholinergic receptors. 3. The secretion elicited through all three receptors appeared to involve exocytosis as revealed by electron microscopy. 4. The beta-agonist, isoprenaline, increased tissue content of cyclic adenosine 3',5'-monophosphate (cyclic AMP); alpha-adrenergic and cholinergic agents had no effect on the level of this cyclic nucleotide. 5. Secretion via cholinergic or alpha-adrenergic mechanisms required extra-cellular calcium; the beta-adrenergic mechanism did not. 6. It was concluded that stimulation of rat parotid cells activates distinctly separate pathways leading ultimately to exocytosis, one pathway involving cyclic AMP, and the other, external Ca2+ ion.


An electrophysiological study of the sacral parasympathetic pathway to the colon of the cat. 1. Electrophysiological techniques were used to study the sacral parasympathetic pathway to the colon of the cat. 2. Electrical stimulation of the sacral ventral roots or the pelvic nerve elicited contractions of the colon and firing in nerve filaments on the serosal surface of the colon. Both responses were markedly reduced by the administration of ganglionic blocking agents. It is concluded that sacral preganglionic fibres to the colon make synaptic contacts with extramural ganglion cells. These cells were identified histologically in small ganglia on the serosal surface of the distal colon and rectum. 3. Transmission in extramural colonic ganglia was cholinergic and mediated by nicotinic receptors. Colonic ganglia did not exhibit large recruiting responses during repetitive (1-4 c/s) preganglionic nerve stimulation or an adrenergic inhibitory mechanism, both of which have been identified in bladder parasympathetic ganglia. It is concluded that colonic ganglia unlike bladder function primarily as simple relay stations and have little potential for modulating the neral activity arising in the central nervus system. 4. The preganglionic input to colonic ganglia was mediated by C fibres with maximal conduction velocities ranging from 0-5 to 1-4 m/sec. Bladder ganglia, on the other hand, received a preganglionic input composed of B fibres with maximal conduction velocities ranging from 8 to 10 m/sec. The possible physiological significance of different types of preganglionic fibres in the sacral outflow is discussed.


Brain stem afferents to the rat medial septum. 1. Activity of neurones in the medial septal nucleus and the diagonal band was recorded from urethane anaesthesized rats. Responses of the cells to electrical stimulation of the raphe nuclei and nucleus locus coeruleus (LC) were measured. 2. LC stimulation caused a long latency, 30-100 msec, and long duration, 100-300 msec cessation of spontaneous activity of most recorded neurones. When bursting-type neurones were recorded, the stimulation occasionally caused a synchronized repetitive bursting firing pattern. 3. Pre-treatment with drugs which interfere with catecholamine neurotransmission, i.e. reserpine and 6OHDA, prevented the appearance of cellular responses to LC stimulation. 4. Stimulation of the dorsal or the median raphe nuclei generated more complex and less clear-cut responses. These included several types of long (20-50 msec) and short (2-5 msec) latency responses. These responses were also accompanied in some cells by synchronized repetitive bursting. 5. Interference with serotonin neurotransmission with PPCA or reserpine reduced the detection of long latency responses. 6. Short latency responses accompanied by evoked field potentials were recorded also after stimulation of dorsal tegmental nucleus. 7. Rates of spontaneous firing cells were augmented after monoamine neurotransmission interruption whereas after fornix lesion, when there is supposedly an increased monoamine innervation of the septum, cells fire at lower rates than normal. 8. It is suggested that noradrenaline and serotonin may serve as neurotransmitters in the medial-septum-diagonal band areas.


Folk beliefs of the Chinese and implications to psychiatric nursing. Each culture has its own ways of coping with problems. The ancient peoples had developed various devices based on their settings. Those devices are not scientific from the modern man's point of view, however, they have been used over and over through thousands of years. The fact that the world populations have been expanding and growing in tremendous speed, and societies have become more and more complex, people in the world have been confronted with more problems than in the ancient days. Perhaps it is time for the modern man to look back to some of the traditionally used practices in order to draw some implications so that the modern practices can be modified and improved. Through such processes of re-evaluating the traditional practices and modifying modern practices, man will be greatly benefited from the past and the present.


Preparing students for physical restraint. While not encouraging the use of physical restrains on mental patients, the author presents a statement of its continuing use and underscores the need for preparing students for a group of traditional procedures that in some ways are increasingly out of step with our times and aesthetically offensive. Two specific clinical indications for restraint are discussed.


Nursing care in a combined adolescent medical-psychiatric unit. In spite of the problems we have encountered in combining adolescent medical and psychiatric patients, we have found this to be a workable model. In summary, several areas are important in making this model functional: staff interest in both medical and psychiatric patients, in-service programs particularly in psychiatry, and open communication between health care givers. Certainly one of the most positive aspects of this unit is that it provides an environment which allows for easier adjustment to the community after hospitalization. Avoidance of such terms as "mental hospital," "crazy person" is less traumatic to the adolescent patient and his family.


Nursing consultation: a clinical specialty. In summary, the Mental Health Nurse Consultant is committed to the belief that nursing staff are competent to implement appropriate mental health care. Because of this the consultant utilizes both resource and process consultation to assist staff in determining care plans and to support a realistic method for delivery. The Mental Health Nurse Consultant interviews clients in order to assess dynamics of behavior and uses this information for care plan development. The direct interview also enables the consultant to investigate client perceptions of Psychiatric Mental Health Intervention and support the utilization of direct involvement by a member of the Consultation and Liaison Team. The Mental Health Nurse Consultant acts as a facilitator for ongoing utilization of existing resources through resource and process consultation to members of the Department of Nursing. Thus, consultation as a clinical specialty offers an additional opportunity for nurses to utilize individual skills and refocus the tasks of nursing within the general hospital.


Aspects of gastrointestinal tumors during the reproductive years. During the reproductive years, tumors of the digestive organs are uncommon in women in the U.S.A. Nevertheless, two predisposing factors have been identified and are discussed in this article. The first of them concerns the benign and malignant tumors of the liver thought to be associated with the long-term use of oral contraceptives. The second is the increased incidence of colon cancer in patients with longstanding, chronic ulcerative colitis. Measures aimed at prevention and early detection are reviewed and discussed.


Aberrant serum enzyme patterns in dermatomyositis associated with hepatoma. A Caucasian male developed florid dermatomyositis documented by serum enzyme elevation, electromyography, and histology of skin and muscle. Serum enzymes, including creatine phosphokinase (CPK), aldolase, glutamic oxaloacetic transaminase (SGOT), and lactic dehydrogenase (LDH), decreased initially during high dose systemic corticosteroid therapy, although profound muscle weakness persisted. Subsequent elevation of serum LDH and SGOT levels during treatment provided a clue to underlying neoplasia. Primary hepatoma with widespread metastases was found at necropsy.


Bis-basic-substituted polycyclic aromatic compounds. A new class of antiviral agents. 7. Bisalkamine esters of 9-oxoxanthene-2,7-dicarboxylic acid, 3,6-bis-basic ethers of xanthen-9-one, and 2,7-bis(aminoacyl)xanthen-9-ones-xanthenes, and -thioxanthenes. 3,6-Bis[2-(dimethylamino)ethoxy]-9H-xanthen-9-one dihydrochloride (4, RMI 10874DA) and 1,1'-(9H-xanthene 2,7-diyl)bis[2-(dimethylamino)ethanone] dihydrochloride (16, RMI 11513DA) were found to prolong survival of mice infected with lethal challenges of encephalomyocarditis (EMC) virus. They were effective by oral as well as subcutaneous administration and showed broad-spectrum antiviral activity. They were selected for preclinical evaluation from the five series of compounds named in the title that were synthesized in analogy to tilorone and related fluorenone derivatives, described earlier. In addition to 4 and 16, compounds 11, 12, 17, and 18 showed high antiviral activity on oral as well as subcutaneous administration. High antiviral activity on subcutaneous admistration was found in the bisalkamine esters 1,2, and 14, the bis(aminoacyl)xanthenes 23 and 26, the bis(aminoalkylene)xanthene 31, the bis(aminoacyl)thioxanthenes 34-40, and the bis-basic ethers of 9-benzylide-nexanthenes 41 and 42. Structure-activity relationships showed a decrease of oral activity with increased length of side chains and increased molecular weight of dialkylamino substituents of 3,6-bis-basic ethers of xanthen-9-one and of 2,7-bis(aminoacyl)xanthenes and-xanthen-9-ones. At least one carbonyl or alkenyl function in conjugation to the xanthene nucleus either at the 9 position of the nucleus or in the side chains is required for high antiviral activity.