N-Aralkyl substitution increases the affinity of adrenergic drugs for the alpha-adrenoceptor in rat liver.

1 The alpha-adrenoceptor of rat liver plasma membranes was studied by use of the specific alpha-antagonist [3H]-dihydroergocryptine ([3H]-DHEC). Catecholamines and adrenergic compounds displayed an order of affinity that is typical of an alpha-receptor. Nevertheless, protokylol, a potent beta-adrenoceptor agonist, exhibited a higher affinity than that of adrenaline for alpha-sites. This result might be due to its bulky substituent on the amino group. 2 Further displacement experiments between [3H]-DHEC and four pairs of drugs differently substituted on the amino group (isoprenaline vs Cc-25, orciprenaline vs fenoterol, AH 3474 vs labetalol, pindolol vs hydroxybenzylpindolol) provided evidence that N-alkyl substitution decreased the affinity for alpha-sites (20 micromolar less than KD less than 200 micromolar), whereas an N-aralkyl one increased the affinity (0.17 micromloar less than KD less than 4.6 micromolar). 3 It is concluded that a substitution on the amino group by a bulky, hydrophobic moiety enhances the affinity of drugs for the alpha-adrenoceptors.


Calcium requirements for electrically-induced release of an endogenous opiate receptor ligand from the guinea-pig ileum.

Calcium requirements for electrically-induced release of an endogenous opiate receptor ligand in the myenteric plexus-longitudinal muscle strip of the guinea-pig ileum were studied. The naloxone-reversible depression of the electrically evoked contraction caused by stimulation at 10 Hz in normal Krebs solution was markedly reduced by decreasing the calcium concentration in the solution. The depression was greatly diminished by increasing the magnesium concentration in the solution. These results show that the electrically-induced release of an opiate-like material requires calcium ions.


A comparison of the facilitatory actions of 4-aminopyridine methiodide and 4-aminopyridine on neuromuscular transmission.

1 4-Aminopyridine methiodide (4-APMI), a quaternary analogue of aminopyridine (4-AP), was tested for neuromuscular facilitatory actions on the chick biventer cervicis and frog sartorius nerve-muscle preparations. 2 In the chick, 4-APMI (10(-4) to 10(-2) M) augmented indirectly elicited twitches and reversed tubocurarine-induced neuromuscular block. Reversal of tubocurarine block was observed after treatment of the muscle with an anticholinesterase. 4-APMI did not itself produce contracture but augmented responses to added acetylcholine. 3 4-APMI (10(-4) M) prolonged the time courses of endplate potentials (e.p.ps) and miniature endplate potentials (m.e.p.ps) in the frog. 4 4-APMI (10(-4) M) increased e.p.p. quantal content. 4-AP was about 100 times more active than 4-APMI in increasing quantal content. Both compounds prolonged muscle action potentials at similar concentrations. 5 4-APMI (10(-3) to 3 X 10(-3) M) possessed anticholinesterase activity in homogenates of chick biventer cervicis muscle. 6 It is concluded that 4-APMI increases evoked acetylcholine release and also possesses a weak anticholinesterase action. The greater action of 4-AP on quantal content is probably due to an intracellular action, possibly involving the release of calcium ions.


Cyclic adenosine 3', 5'-monophosphate and the mechanism of action of three common anti-inflammatory drugs.

1 The effects of indomethacin, dexamethasone and colchicine on cyclic adenosine 3', 5'-monophosphate (cyclic AMP) concentration in leucocytes during a crystal-induced pleurisy in rats were studied. 2 Each of the drugs significantly increased leucocyte cyclic AMP content within 3 h of the injection of crystals. 3 By 6 h, leucocyte cyclic AMP levels returning toward control levels and could not be sustained at the higher level by an additional administration of the respective anti-inflammatory drug.


alpha And beta-adrenoceptors in the detrusor muscle and bladder base of the pig and beta-adrenoceptors in the detrusor muscle of man.

1 The presence and type of adrenoceptors in the smooth muscle of the pig and human urinary bladder was assessed on the basis of the relative potency of alpha- and beta-adrenoceptor agonists and antagonists.2 In isolated, carbachol-contracted bladder strips from the pig detrusor muscle the relaxing potency of isoprenaline was four times that of salbutamol and ritodrine and thirty times that of noradrenaline.3 Propranolol caused a parallel shift to the right of the noradrenaline dose-response curve which was not changed by phentolamine.4 Propranolol and butoxamine showed, in contrast to practolol, a dose-dependent antagonism of the response to isoprenaline. A pA(2) value of 9.2 +/- 0.2 and 6.8 +/- 0.2 (mean +/- s.e. mean) for the first two antagonists was calculated.5 In the bladder base of the pig, propranolol caused a parallel shift to the right and phentolamine a shift to the left of the dose-response curve to noradrenaline.6 In the human detrusor muscle the potency and maximum effect of isoprenaline and salbutamol were less than those in the pig detrusor muscle. The potency of isoprenaline was sixty times that of salbutamol.7 Whereas a parallel shift to the right of the dose-response curve to isoprenaline was obtained with propranolol, no antagonism was obtained with butoxamine or practolol.8 The results are interpreted as indicating the presence of beta(2)-adrenoceptors in the detrusor muscle of the pig and beta-adrenoceptors with neither beta(1)- nor beta(2)-characteristics in the detrusor muscle of man. An indication of the presence of alpha-adrenoceptors in the bladder base but not in the detrusor muscle of the pig was obtained.


Comparison of pre-junctional alpha-adrenoceptors at the neuromuscular junction with vascular post-junctional alpha-receptors in cat skeletal muscle.

1 Activation of pre-junctional alpha-adrenoceptors at the skeletal neuromuscular junction enhances acetylcholine release whereas activation of such receptors at autonomic nerve endings inhibits transmitter output. In the present study the characteristics of pre-junctional alpha-adrenoceptors at motor nerve terminals have been compared with post-junctional (vascular) alpha-adrenoceptors in the cat hind limb.2 Reversal of partial (+)-tubocurarine blockade of contractions of the tibialis anterior muscle was used to monitor pre-junctional activity and increases in hindlimb vascular resistance to assess post-junctional actions at alpha-adrenoceptors.3 Responses to intra-arterial injections of noradrenaline, adrenaline, phenylephrine, oxymetazoline, methoxamine and clonidine were monitored. Dose-response lines for all the compounds except clonidine were parallel. The latter agent produced only weak and inconsistent effects.4 Ratios of the doses of the agents required to produce pre- and post-junctional effects indicated that oxymetazoline and adrenaline possessed some preferential activity at post-junctional sites, whereas the remaining agents were non-selective in their actions. If dose-ratios with respect to noradrenaline were compared at the two sites none of the compounds possessed a marked degree of selectivity.5 In the presence of phentolamine or tolazoline, dose-response curves to the pre- and post-junctional effects of phenylephrine were shifted to a similar extent. Thymoxamine showed preferential activity as a pre-junctional alpha-receptor antagonist.6 In comparing the results of this study with those of other authors, it is apparent that there are marked differences in the characteristics of pre-junctional alpha-receptors at the skeletal neuromuscular junction and at autonomic nerve endings. The pre- and post-junctional alpha-receptors in skeletal muscle show less divergence.


Depolarization of neurones in the isolated olfactory cortex of the guinea-pig by gamma-aminobutyric acid.

1 Effects of gamma-aminobutyric acid (GABA) on single neurones in slices of guinea-pig olfactory cortex maintained in vitro were recorded with single intracellular microelectrodes. The average resting potential of 52 cells was -75 mV and apparent input resistance ranged from 20 to 200 MOmega.2 Superfusions of GABA over the slice invariably depolarized the neurones and reduced their input resistance. The minimum effective concentration was 50 to 200 muM.3 The reversal potential for the depolarization produced by 0.1 mM GABA (E(g)) was -66 +/- 2 mV. At concentrations >0.1 mM the reversal potential became progressively more positive (-55 to -50 mV).4 Reduction of external chloride, with isethionate as the substitute anion, increased the amplitude of the depolarization.5 GABA reduced the amplitude of the excitatory postsynaptic potential produced by lateral olfactory tract stimulation, and occluded or reversed the subsequent depolarizing recurrent inhibitory postsynaptic potential.6 Action potentials elicited by injection of depolarizing current or by focal antidromic stimulation were slowed and reduced in amplitude by GABA.7 The effects of GABA on membrane conductance (potency = 1) were duplicated by 3-aminopropanesulphonic acid (potency = 20), beta-alanine (0.5), beta-amino-n-butyric acid (0.5), glycine (0.3) and L-2,4-diaminobutyric acid (0.2). For a given conductance change, 3-aminopropanesulphonic acid, glycine and beta-alanine produced less depolarization than did GABA.8 It is concluded that the action of GABA on the neurones is compatible with a role in mediating recurrent postsynaptic inhibition.


Selective blockade by scopolamine of synaptic responses in cat's caudate nucleus and its modification by lesions of the substantia nigra.

Because it is commonly believed that acetylcholine is a synaptic transmitter in the caudate nucleus and that the reduction of striatal biogenic amines in Parkinson's disease leads to acetylcholine supersensitivity in the caudate nucleus, we investigated the effects of the muscarinic blocking agent scopolamine on synaptic responses of neurons in the intact feline caudate nucleus and in the caudate nucleus depleted of dopamine by long-standing nigrostriatal lesions. In the intact caudate nucleus, micro-iontophoretic application of scopolamine selectively blocked the neuronal responses to stimulation of the caudate nucleus near the recording site without affecting the responses to stimulation of the sensorimotor cortex or the substantia nigra in the same fashion. This suggests that acetylcholine is a synaptic transmitter of caudate interneurons. Responses to thalamic stimuli were also blocked by scopolamine, suggesting that acetylcholine may be a transmitter of thalamic afferents although the course of these afferents is unclear. In the dopamine-depleted caudate nucleus scopolamine was more effective than in the intact caudate nucleus blocking the neuronal responses to stimulation of the caudate nucleus. This greater blocking effect by scopolamine suggests an increased effect of endogenous acetylcholine in this response and supports previous observations of an increased excitatory effect of iontophoretic acetylcholine in the dopamine-depleted caudate nucleus. These results suggest that the acetylcholine supersensitivity which follows nigrostriatal degeneration may be due to increased effectiveness of synaptic transmission by cholinergic interneurons in the caudate nucleus.


Responses of single units in laminae 2 and 3 of cat spinal cord.

333 units were recorded in laminae 2 and 3 of lumbar cord in decerebrate cats. Recording of small-amplitude spikes was made possible by the use of platinum-surfaced tungsten microelectrodes, continuously variable filters and an analogue delay line display. Stimulation of the lateral Lissauer tract showed that a sample of the units sent axons into this tract. Using iron-plated electrodes, recording sites were marked and shown to be within laminae 2 and 3. Axons of peripheral afferent axons were excluded from the sample, as were long-range descending axons. By using one electrode placed close to a cell body in lamina 4 and a roving electrode in the dendritic region dorsal to the cell body, it was possible to show that the recorded units were not field spread of deeper cells. Of the units 94% had peripheral receptive fields, RF; 30% had small RFs less than 2 sq.cm, intermediate in size between RFs of peripheral axons and RFs of large cells in dorsal horn. These small RF cells occurred in clusters and their RFs constituted a fraction of the larger RF of nearby large cells. Of the units 56% responded to brush and touch, 19% to brush, touch and pressure, while 19% required pressure on skin to excite them. Latency of response to electrical stimulation showed that all cells were excited by myelinated afferents. While no cells were detected exclusively by C afferents, many may have been excited by both A and C afferents. Eighteen per cent of the cells showed a prolonged discharge lasting more than 5 sec after a single stimulus. Some of these long discharge cells continued firing for minutes. Another unusual class, 14% of all cells, habituated very powerfully to intermittant natural or electrical stimuli, and remained unresponsive for many seconds after responding to the first stimulus.


Opiate-like excitatory effects of steroid sulfates and calcium-complexing agents given cerebroventricularly.

Intracerebroventricular administration of 10--20 microgram of steroid-O-sulfates induced hypermotility, agitation, salivation, EEG abnormalities, stereotypies, wet dog shakes and seizures. Equivalent effects resulted from 30--200 microgram morphine sulfate (H2SO4 salt), 50 microgram EGTA or 300--400 microgram of sodium sulfate or phosphate, but not chloride, nitrate or acetate. Non-steroid sulfates, steroid glucuronides and steroid phosphates were inactive. Naloxone, previously found to antagonize the excitatory effects of androsterone sulfate, failed to antagonize those of cortisol sulfate, sodium sulfate or EGTA. These findings suggest a role for extracellular calcium ions and for sulfate derived from circulating steroids in central responses to opiates.


Analysis of the habituation-like changes in transmission in the temporodentate pathway of the rat.

Habituation-like decrements in extracellular measures of synaptic activation (population EPSP) and cell discharge (population spike) were analyzed in the dentate gyrus of the rat following repetitive low-frequency stimulation of the medial and lateral entorhinal cortex. Stimulation of either subdivision of the entorhinal projection system resulted in comparable habituation-like response decrements with similar stimulation regimens. However, habituating stimulation of one subdivision did not result in decreased responsiveness to stimulation of the other. Repetitive low-frequency stimulation or even a single pulse delivered to either subdivision did, however, result in a potentiation of granule cell discharge in response to stimulation of the other subdivision (a form of heterosynaptic potentiation). This heterosynaptic potentiation of granule cell discharge was not accompanied by any increase  in the extracellular EPSP. Comparisons of the relationship between the population EPSP and population spike before and during habituating stimulation revealed changes in cell discharge in response to the habituating stimulus which could not be accounted for by changes in synaptic activation alone. The results suggest that repetitive activation of the temporodentate pathway alters granule cell output as a result of two processes, a habituation-like decrement in synaptic activation, and a potentiation of granule cell discharge as a consequence of prior activation.


The effect of hypothalamic hemi-islands on compensatory adrenal growth.

Unilateral hypothalamic disconnections were made with a Halász knife in young male rats to determine the effects of these lesions on adrenal weight, and the response to unilateral adrenalectomy. Rats were unilaterally adrenalectomized or sham-adrenalectomized 5-7 days after the hypothalamic surgery. No lesion affected adrenal weight in sham-adrenalectomized rats. Compensatory adrenal growth 3 days after unilateral adrenalectomy is prevented by a unilateral hypothalamic hemi-island on the side ipsilateral, not contralateral, to the first removed adrenal (P less than 0.01). Portions of the ipsilateral hemi-island were examined to determine which interrupted the compensatory adrenal growth response. The anterior 90 degrees portion enhanced compensatory adrenal growth (P less than 0.01). The posterior 90 degrees portion only partially inhibited compensatory adrenal growth (P less than 0.05), while the posterolateral portion completely inhibited the response (P less than 0.01). Plasma ACTH and corticosterone levels at the time of sacrifice were unaffected by unilateral adrenalectomy, but were slightly elevated by the hypothalamic cuts. These studies provide further evidence that compensatory adrenal growth is mediated neurally.


Spontaneous phagocytosis of C-type synaptic terminals by spinal alpha-motoneurons in newborn kittens. An electron microscopic study.

The cell bodies and proximal dendrites of spinal alpha-motoneurons were studied electron microscopically with the aid of serial sections during the first postnatal week in the cat. The observations suggested that some of the synaptic terminals of the so-called C-type on the cell bodies and dendrites are phagocytosed by the motoneurons during the first few days after birth. This finding is discussed in relation to the earlier demonstrated postnatal loss of synaptic terminals on the motoneurons after birth and differences demonstrated between different functional types of spinal motoneurons with respect to the number and distribution of C-type terminals in the adult cat.


Postsynaptic potentials recorded from medullary neurones following stimulation of carotid sinus nerve.

Projections of the carotid sinus nerve (CSN) onto medullary neurones were studied with intracellular recording. Three types of postsynaptic potentials (EPSP, EP-IPSP and IPSP) were recorded by stimulation of the ipsilateral CSN. Of the total of 121 neurones, positions of 54 were identified by intracellular dye. The other 67 were positioned by extrapolation. They were distributed over 5 medullary nuclei: (1) nucleus of the solitary tract (NTS); (2) paramedian reticular nucleus (NPR); (3) perihypoglossal nucleus (PXII); (4) lateral tegmental field (FTL); and (5) nucleus ambiguus. Since penetration of microelectrodes and injection of dye into the NTS neurones was difficult, neurones of the other 4 nuclei were examined. The IPSPs were dominant in small NPR neurones, while the EPSPs were dominant in large neurones of the other 3 nuclei. Both the NA and PXII neurones showed forms of a motor type neurone, while the FTL neurones showed various forms. The EPSPs with onset latency as short as 2-4 msec were frequently recorded in different nuclei. This strongly suggests that the CSN projects monosynaptically onto different nuclei in the medulla.


Two pharmacologically distinct histamine receptors mediating membrane hyperpolarization on identified neurons of Aplysia californica.

Two distinct hyperpolarizing responses are produced when histamine is iontophoretically applied onto the somal membranes of identified neurons within the cerebral ganglion of Aplysia: a biphasic response consisting of a rapid component (less than 5 sec) usually superimposed upon a slowly developing component; or a monophasic slowly developing response 5-20 sec in duration. The reversal potential values for the fast (typically -65 mV) and the slow (typically -89 mV) responses, and their shift to new values when the external potassium or chloride concentrations were altered, revealed that the fast and slow potentials are produced predominantly by conductance increases to chloride and potassium ions, respectively. The effects of histamine H1- and H2-receptor agonists and antagonists were studied to characterize the pharmacological properties of histamine receptors mediating these two ionically dissimilar hyperpolarizing responses. The slow potassium-dependent hyperpolarization could be mimicked by several histamine analogues; the most potent tested were the H1-receptor agonist, 2-methylhistamine, and the H2-receptor agonist, 4-methylhistamine. Neither of these agents mimicked the fast chloride-dependent histamine response. The slow potassium-dependent responses induced by histamine or histamine agonists were completely and reversibly blocked by the H2-receptor antagonist, cimetidine. By contrast, the slow potassium-dependent hyperpolarizations produced by iontophoretically applied acetylcholine or by dopamine to the same neurons were unaffected by cimetidine. Other H1 and H2 antagonists tested were either ineffective, or only partially blocked the slow hyperpolarizations in a non-selective manner. The fast chloride-dependent hyperpolarizations were not selectively antagonized by any of the H1 or H2 reagents tested, although they were effectively suppressed by tubocurarine and strychnine. These data indicate that two pharmacologically distinct histamine receptors mediate potassium- and chloride-dependent hyperpolarizations in Aplysia neurons. Neither of these receptors, however, could be classified as strictly H1 or H2 according to criteria presently used in non-neuronal tissues. The selectivity and reversibility of cimetidine indicate that this particular antihistaminic could be a valuable pharmacological tool for defining putative histaminergic synapses in Aplysia and perhaps other nervous systems.


Divalent cation dependent phosphorylation of proteins in squid giant axon.

In vitro and in situ (after intracellular infusion) incubation of axoplasm from the squid giant axon with [gamma-32P]ATP produces a phosphorylation of primarily two proteins (of mol.wt. 200,000 and greater than 400,000). The phosphorylation of these proteins is stimulated by Mg2+, inhibited by Ca2+, and unaffected by 10(-7) to 10(-5) M cyclic nucleotides. The 200 kdalton and greater than 400 kdalton phosphorylated peaks appear to be neurofilament proteins, and phosphorylation of these peaks in situ is decreased by electrical stimulation of the axon.


Effects of vitamin D3 metabolites on bone cell calcium transport.

The vitamin D3 metabolite, 25-hydroxycholecalciferol, at concentrations of 0.01 to 10.0 microgram/ml, decreased calcium uptake by isolated bone cells. The effect occurred within 1 min after the simultaneous addition of metabolite and 45Ca. Lactic acid and ATP production by the cells was not affected. 24(R), 25-dihydroxycholecalciferol produced a similar decrease in calcium uptake. Vitamin D3 had no effect at concentrations from 0.01 to 10.0 micrograms/ml. No effect of 1,25-dihydroxycholecalciferol on calcium uptake was observed with concentrations from 0.1 to 100 ng/ml and various preincubation periods extending to 2 h. None of the agents had any effect on calcium efflux. The effects of 25-hydroxycholecalciferol and 24(R), 25-dihydroxycholecalciferol on calcium uptake were not seen in isolated fetal rat skin cell preparations.


Hydroxyl radical production by free and DNA-bound aminoquinone antibiotics and its role in DNA degradation. Electron spin resonance detection of hydroxyl radicals by spin trapping.

The reduced antitumor antibiotic mitomycin C in aqueous solution exposed to air gives a 36-line electron spin resonance spectrum of the semiquinone identified by computer simulation. Incubation of this radical with the spin trap N-tert-butyl-alpha-phenylnitrone (PBN) gives the PBN.OH nitroxide radical identified by independent generation. This nitroxide radical is also formed from similar treatment of a DNA to which mitomycin C is covalently attached. Incubation of the semiquinone from mitomycin C, mitomycin B, or streptonigrin (SN) with catalase or with superoxide dismutase inhibits the generation of OH, implying the intermediacy of H2O2 and O2 in its formation. The formation of the spin-trapped nitroxide radical is similarly inhibited by EDTA, suggesting the intermediacy of trace metal ions in the generation of hydroxyl radicals from SN. The results are consistent with the generation by the aminoquinone antibiotics in vivo of OH. already implicated in the degradation of DNA.


Case report: An outbreak of herpesvirus myeloencephalitis in vaccinated horses.

In the foaling season of 1977, five vaccinated horses in a Standardbred breeding stable were affected with herpesvirus myeloencephalitis. Respiratory and abortigenic forms also occurred in other individuals on the premises. Equine herpesvirus type 1 was isolated from the brain of one case of myeloencephalitis and from lungs of two aborted fetuses. Twelve of 16 horses demonstrated fourfold or greater increases in titres to equine herpesvirus type 1.


The role of genetic factors in the etiology of Wilms' tumor: two pairs of monozygous twins with congenital abnormalities (aniridia; hemihypertrophy) and discordance for Wilms' tumor.

Wilms' tumor was diagnosed in two children each of whom has an identical twin. In one of the pairs of twins the aniridia syndrome with psychomotor retardation was present in both children, but Wilms' tumor was found in only one. In the other twins hemihypertrophy as well as Wilms' tumor were identified in one child, whereas neither of these abnormalities was present in her twin sister. These findings lend support to a hypothesis that the development of Wilms' tumor requires the occurrence of two successive mutational events, one of which may be a germinal mutation. The presence of aniridia, hemihypertrophy, or other associated congenital abnormalities may aid in distinguishing between hereditary and sporadic forms of Wilms' tumor.


Atypical fibrous histiocytoma with myxoid stroma: a rare lesion arising from dura mater of the brain.

Intracranial fibrohistiocytic tumors are rare. This report is concerned with a 24-year-old white male who had an 8 month history of intermittent visual disturbance. Craniotomy revealed a large, completely intradural lesion on the floor of the left anterior cranial fossa without involvement of the leptomeninges or brain parenchyma. Gross and microscopic examination revealed that the fibrogenic portion of the tumor was composed of spindle shaped neoplastic cells arranged in storiform pattern, a hallmark of fibrohistiocytic tumor. In contrast to three previously reported cases of "fibrous xanthomas" involving the leptomeninges and superficial cortex of the brain, the present lesion is clearly originated from mesenchymal stem cell of the dura. The other distinct gross and microscopic feature was the presence of myxoid component which constituted about 40% of the entire lesion. The myxoid component had not been observed in previous 5 reported cases of intracranial fibrohistiocytic lesions. Since there was cellular pleomorphism with extremely rare mitosis, it was felt that the lesion be best designated as atypical fibrous histiocytoma with myxoid stroma.


Reactive glioma in intracranial sarcoma: a form of mixed sarcoma and glioma ("sarcoglioma"): report of eight cases.

The clinicopathologic features of eight new cases of combined intracranial sarcoma and glioma are described. This type of mixed cerebral tumor is histologically characterized by a peripheral distribution of the gliomatous elements in relation to a more centrally situated meningeal or intracerebral sarcoma, and by the frequent presence of gradual transitions from reactive to frankly neoplastic astrocytes. In six of the eight cases, the additional development of either infiltrating astrocytoma or frank glioblastoma in the adjacent brain was demonstrated; this was interpreted as a further expression of malignant glial reaction. It is suggested that these tumors be termed "sarcogliomas" to distinguish them from the type of mixed glioma and sarcoma that has recently been redesignated "gliosarcoma."


Metastatic carcinoma to the tongue: a report of two cases and a review of the literature.

Metastatic tumors of the oral cavity are uncommon. Most of them are located in the mandible while only a small percentage are found within the soft tissues. Two cases of metastatic bronchogenic carcinomas to the tongue are described. Review of the literature showed that cancer of the lung was the most common primary lesion that metastasized to the soft tissues of the oral cavity. The base of the tongue was the most prevalent site of metastases to soft tissue within the oral cavity.


Systemic leishmaniasis mimicking malignant histiocytosis.

A 22-year-old man with fever, hepato-splenomegaly and severe pancytopenia is described in whom the histologic features of the spleen, liver and lymph nodes were consistent with malignant histiocytosis. Characteristic Leishman-Donovan bodies were demonstrated on a bone marrow aspirate, and the diagnosis of visceral leishmaniasis was confirmed by culturing the flagellated forms of the pathogen, and by an excellent response to sodium stibogluconate. In view of the similarity in clinical and histologic appearance, visceral leishmaniasis must be considered in the differential diagnosis of malignant histiocytosis in geographic areas where leishmaniasis is still endemic.


Multiple pulmonary lesions in a patient treated with BCNU (1,3-bis(2-chloroethyl)-1-nitrosourea) for glioblastoma multiforme.

A patient with a glioblastoma multiforme who was treated with BCNU, developed rapidly progressive pulmonary disease. The pulmonary changes were studied pathologically and included atypical alveolar cell hyperplasia, hyaline membranes, and interstitial fibrosis. These lesions contributed to the patient's demise and may be related to the administration of BCNU.


Detection of humoral immune response to human brain tumors: specificity and reliability of microcytotoxicity assay.

A humoral microcytotoxicity assay has been used to study the immune response to human malignant astrocytic tumors. Two series are described. When the assay was performed under uniform, ideal conditions, the sera from 36 of 45 patients with these tumors (80%) produced significant cytotoxicity against allogeneic astrocytoma cells. Only 4 of 55 (7%) normal sera tested demonstrated cytotoxicity. The second series describes a five year study of over 400 sera with several sera assayed each week. The assay was found to vary in specificity and reliability over this five year period. The reasons for this variability are discussed and methods of standardizing the technique are proposed. Recent information concerning the mechanisms of the humoral microcytotoxicity assay offer promise of improved reliability and clinical utility.


Inhibition of sodium-potassium-activated adenosine 5'-triphosphatase and ion transport by adriamycin.

The antitumor antibiotic Adriamycin is a potent inhibitor of the sodium-potassium-activated adenosine triphosphatase of native heart microsomes. Adriamycin also inhibits potassium transport (although not sodium transport) in slices of kidney cortex. The effects on both the adenosine triphosphatase and ion transport are markedly reduced by Ca2+, probably by chelation of this metal by Adriamycin. These effects could provide a basis for explaining the Adriamycin cardiotoxicity as a digitalis-type toxicity.


Differences in the ability of human lymphoblastoid lines to exclude bromodeoxyuridine and in their sensitivity to methyl methanesulfonate and to incorporated [3H]thymidine.

Six Burkitt's lymphoma-derived lines were more resistant to methyl methanesulfonate than were six lymphoblastoid lines derived by in vitro transformation as measured by cloning ability in soft agar. The differences were due to cell response rather than to reactivity. Lymphoblastoid lines were also more sensitive to [3H]thymidine incorporated along with bromodeoxyuridine than were the lymphoma lines tested. All lines showed excision repair activity as measured by the benzoylated napthoylated DEAE cellulose method, although there were quantitative differences in the activity of different lines. Most lymphoblastoid lines synthesize DNA of lower density when incubated in analog-containing medium due to their ability to discriminate against bromodeoxyuridine and to select for thymidine in a mixture. All the lymphoma cell lines tested were unable to discriminate between thymidine and bromodeoxyuridine. The ability of lymphoma cells to produce large quantities of dense radioactive, doubly substituted DNA when incubated in medium with bromodeoxyuridine and thymidine is due to their inability to discriminate between nucleosides and to their resistance to incorporated [3H]thymidine.


Imidoester inhibition of lymphocyte DNA synthesis.

Imidoesters amidinate free amino groups and produce inter- and intramolecular covalent bonds. To determine whether imidoesters influenced lymphocyte transformation, human peripheral blood or calf lymph node lymphocytes were cultured with dimethyladipimate (DMA), a bifunctional (cross-linking) imidoester, or methyl acetimidate (MAC), a monofunctional (noncross-linking) imidoester. Both DMA and MAC decreased the rate of endogenous DNA synthesis in a dose-dependent fashion. In further work, lymphocytes were treated with Phaseolus vulgaris phytohemagglutinin, concanavalin A, or periodate. DMA (1 mM) decreased DNA synthesis in P. vulgaris phytohemagglutinin-stimulated human cells by 65%, Concanavalin A-stimulated cells by 98.2%, and periodate-stimulated cells by 85%. Similar results were obtained with 1 mM MAC. Inhibition by DMA was slightly greater than was the inhibition by MAC. Decreased DNA synthesis resulted if DMA was added to P. vulgaris phytohemagglutinin-stimulated human lymphocytes at initiation of culture (72%) or after 16 hr (75%); inhibition was less when DMA was added after 24 hr (43%) and was not apparent if added after 48 hr. Therefore, both monofunctional and bifunctional imidoesters inhibit endogenous and stimulated DNA synthesis in human and calf lymphocytes.


Inhibition of human lymphocyte mitogen and antigen response by a 15,000-dalton protein from feline leukemia virus.

Peripheral blood lymphocyte response of normal human subjects to mitogens and antigens was suppressed by a 15,000-dalton protein (p15) from a C-type feline leukemia virus. Four of six subjects were suppressed 70 to 96% when responding to concanavalin A or phytohemagglutinin in the presence of 5.0 microgram of p15. The three subjects who responded to streptokinase-streptodornase and Candida were suppressed 68 to 91% when cultured with 5.0 microgram of p15. The subviral protein did not appear to be cytotoxic at doses reported. Lymphocyte membrane studies with fluorescein isothiocyanate-concanavalin A revealed a reduction in concanavalin A-induced cap formation of 51 to 91% in the presence of the same dose of p15. These results demonstrate that in vitro immunological dysfunction in human lymphocytes can be induced by a C-type virion protein.


Transplantable ductal adenocarcinoma of the Syrian hamster pancreas.

A well-differentiated ductal adenocarcinoma of the Syrian golden hamster induced by N-nitrosobis(2-oxopropyl)amine was transplantable to both nude mice and inbred Syrian hamsters. The tumor grew rapidly in the nude mouse (12-fold increase in size at 45 days) in contrast to its growth in hamster (3-fold increase in size at 45 days). A curious finding associated with the slow-growing tumor in the hamster was an intense infiltration of the neoplasm by polymorphonuclear leukocytes unattended by either necrosis or infection. The neoplasm produced mucin and rapidly and specifically bound 125I-labeled secretin, although the degree of nonspecific binding (40.5%) was higher than that of control hamster pancreas (23%). Unstimulated adenyl cyclase activity (pmol cyclic adenosine 3':5'-monophosphate per mg protein) of the neoplasm was significantly higher [3.76 +/- 0.55 (S.E.)] than that of unstimulated normal hamster pancreas (1.03 +/- 0.44). Secretin did not significantly change the level of cyclic adenosine 3':5'-monophosphate (3.3 +/- 0.56) from the unstimulated level in the neoplasm, in contrast to its effect on normal pancreas where the level was increased 3-fold (3.1 +/- 0.75).


Cross-reactivity between Bacillus Calmette-Guérin and Rous Virus-induced sarcoma detected in rats by tube leukocyte adherence inhibition assay.

Specific immunity to Rous virus-induced sarcoma (RSL) was investigated by the tube leukocyte adherence inhibition (LAI) assay in rats immunized with Bacillus Calmette-Guérin (BCG). Peritoneal cells from Lewis rats immunized s.c. with BCG gave positive reactions in the tube LAI assay with the antigen prepared from RSL in Lewis rats. Lymph node cells from Lewis rats immunized with BCG had no cytotoxic effect on RSL cells in vitro, whereas peritoneal cells from the same rats were strongly cytotoxic for RSL cells. Growth of RSL tumors in vivo was not inhibited in BCG-treated rats as compared to that in untreated rats. The results show that LAI reactivity correlates with cytotoxicity of peritoneal cells and does not correlate with the cytotoxicity of lymph node cells and that positive LAI reactions with tumor antigen need not be reflected in the suppression of growth of the tumor in vivo.


Control of regional circulations by the sino-aortic reflexes during desynchronised sleep in the cat.

In the intact cat desynchronised sleep induces arterial hypotension and mesenteric vasodilatation, whereas in limb muscles vasoconstriction occurs because of the concurrent action of a spinal vasoconstriction reflex. After sino-aortic denervation arterial hypotension is exaggerated and mesenteric vasodilatation is greater, while the muscular bed of the limbs also becomes vasodilated rather than constricted. It is concluded that during desynchronised sleep the sino-aortic afferents act to check a central influence inhibiting vasoconstrictor tone both in the viscera and muscles: thus an excessive fall in blood pressure is prevented by avoiding a too large visceral vasodilatation and by unmasking the vasoconstrictor action of the spinal reflex in the limbs.


The effect of cyclic AMP on the growth and morphology of a normal human fibroblast parent strain and its transformed progeny line.

A normal human diploid fibroblast cell strain, Lederle 130 (Led 130), and its virus-transformed progeny line, transformed Led 130, were subjected to 0.75 and 1.5 mM concentrations of adenosine-5'-monophosphate (AMP), cyclic AMP (cAMP) and dibutyryl cyclic AMP (Bt2cAMP). While cAMP was markedly inhibitory to neoplastic cells at 1.5 mM, Bt2-cAMP was even more effective at this concentration, producing 85% inhibition by 4 days and 91% inhibition by 6 days. Bt2-cAMP was the only nucleotide to reverse morphological transformation effects in the neoplastic fibroblasts. Normal fibroblasts were inhibited in growth rate to a comparable extent by all nucleotides, and were not altered morphologically.


[Toxicity of silica-gel polymers].

Mice of the ICR strain were tested for the effect of intraperitoneally applied polymers of silicagel. Silicagel applied in the form of spherical-shaped particles caused local irritation and resorptive reaction with granulocytes. There was also renal damage of the type of necronephrosis and signs of necronephrosis.


[Changes following splenoportography or lymphography, respectively].

Lipidol deposits remaining in lymph nodes following lymphography evoke a tissue reaction of various degree. Most probably, the material injected during splenoportography may lead to storage phenomena in the reticuloendothelium of the liver.


Isolation of viruses from sewage, with special regard to poliovirus: an evaluation of the sensitivity of three tissue culture methods.

This report concerns experiments to isolate different viruses from sewage. Using a special cell-line from Utrecht, derived from human amniotic cells, it was possible to isolate poliovirus selectively when antisera against six types of coxsackievirus B were added to the tissue culture. The method was tested in connexion with the epidemiological investigation of a case of poliomyelitis in Sweden in 1977. It rapidly demonstrated that the virus implicated was present in all neighbouring sewage plants, indicating a wide distribution of the virus in the area.


[Muco epidermoid tumor of the bronchus in a 8 year old girl (author's transl)].

A 8 year-old girl was referred with a mucoepidermoid tumor of the bronchus. Surgery was tried before a mediastin left likeness; diagnosis resided in histology of the surgical piece. Reading of medical literature, very slight for children, corroborate difficulties of histologic classification of these tumors, and especially their doubtful forecast.


Serum cholesterol and squalene in hepatocellular carcinoma.

In 15 patients with hepatocellular carcinoma, the serum squalene level is raised significantly in parallel with the serum cholesterol level, though significantly decreased in patients with cholestatic hepatobiliary disease. Serum levels of cholesterol and/or squalene, therefore, could be regarded as a laboratory index reflecting a peculiar sterol metabolism in hepatocellular carcinoma.


Cyclic alterations of blink reflexes: an EEG and EMG study during wakefulness and sleep.

Studies of the blink reflex were undertaken in twelve normal subjects during wakefulness and different sleep stages. The early R1 and the late R2 components of the reflex were analyzed. R1 was present in only one subject during stage I sleep and subsequently disappeared in stage II to IV. In all subjects, R2 was gradually diminished and totally disappeared during stage IV. During the REM stage, R1 was seen in only four-out-of-twelve subjects but all R2 returned. Alterations of R1 and R2 of the blink reflex during different sleep stages suggest that the underlying mechanism includes both supranuclear and infranuclear activities. Our finding strengthens further the notion of dual hyponogenic mechanisms for sleep.


Stimulation of capillary tube polymorph migration: an indirect glucocorticoid effect on microtubular function.

Polymorph migration stimulator (PMS) is a peptide factor produced by an in vitro reaction between glucocorticoids and human mononuclear phagocytes. This study was undertaken to determine the significance of the stimulatory effect of PMS on the capillary tube migration of human polymorphs. Colchicine, vinblastine and Nocodazole, all of which inhibit microtubular assembly, are shown to stimulate migration. Conversely, deuterium oxide which stabilizes microtubules inhibits migration. Increased intracellular cyclic AMP is associated with microtubular inhibition and isoprenaline, theophylline and dibutyryl cyclic AMP are also found to stimulate capillary tube migration. These results suggest that PMS acts by inhibiting the assembly of polymorph microtubules, an effect which may be mediated by cyclic AMP in the same manner as other peptide hormones.


Plasma levels of complement components and complement haemolytic activity in protein-energy malnutrition.

The plasma levels of complement haemolytic activity (CH50) of some complement components and of C3d, a C3 breakdown product, were measured in fifty-nine African children with various types of protein-energy malnutrition (PEM) including kwashiorkor, before and during recovery. A significant decrease of CH50, C3, C9 and factor B was observed in PEM without a concomitant decrease of C4 and C5. Increased plasma levels of C3d were also found in PEM patients. Two mechanisms seem to be involved in the impairment of the complement system in PEM: (1) a decreased synthesis of at least C3 and C9 as suggested by a significant correlation between C3 or C9, levels and those of albumin and cholinesterase; (2) an increased catabolism of C3 possibly due to an activation of the alternative complement pathway, as suggested by the increased level of C3d and the decreased level of factor B which are significantly correlated with C3 levels but not with albumin levels. These data support the possible role of a relative complement deficiency upon the decreased resistance to infections observed in malnourished children.


Adipose tissue cellularity in patients with amyotrophic lateral sclerosis.

The subcutaneous mean fat-cell volumes as measured in 20 patients suffering from amyotrophic lateral sclerosis (ALS) were definitely larger than those measured in a control group. In contrast with the control subjects, the mean fat-cell volume in patients with ALS appeared to be independent of body weight. The noted disturbances in carbohydrate-insulin metabolism in patients with ALS and the increase in serum triglycerides as observed in some patients may be related to the enlarged fat cells. The possibility that the enlargement of the fat-cells may, at least partially, have a neurogenic basis cannot be ignored. The hypothesis is put forward that in patients with ALS there is not only a lesion of the anterior horn, but also an involvement of sympathic structures responsible for the innervation of adipose tissue vessels. This involvement may lead to an inhibition of the lipolysis, and consequently to an enlargemnt of the fat cell. Interaction between the interference with metabolic and neurogenic factors may be at play.


Programmed texts: success or failure? An analysis of medical students' opinions.

It is shown that German medical textbooks have sold about nine times better than programmed texts, even if they were written by the same author and on the same subject. To explore some of the reasons why programmed texts are being used so little in comparison with conventional textbooks, 4th and 5th year medical students of the University of Berne were asked to work through a short programmed text in clinical neurology and to answer questions related to several aspects of using programmed texts and other materials. The analysis of the 212 questionnaires returned (58%) indicates that almost all students considered such a learning experience as highly useful and stimulating, but only about 25 per cent had no reservations. It is concluded that programmed medical texts may be used so little because they require students to learn in a highly standardized way, and because students need additional learning resources which provide them with  conceptual framework of and sufficient information about a given area.


Reduced adenosine 3',5'-cyclic monophosphate levels in patients with reversible obstructive airways disease.

1. Patients were grouped into categories of 'no airways disease', 'obstructive airways disease without response to bronchodilator' and 'obstructive airways disease with bronchodilator responsiveness'. 2. Cyclic nucleotides were assayed in specimens of lung tissue that were excised during surgery. 3. Reduced levels of adenosine 3',5'-cyclic monophosphate (cyclic AMP) were found in pulmonary tissue obtained from patients with reversible obstructive airways disease, lending support to the beta-adrenergic theory of asthma.


Computerized tomography in patients with multiple sclerosis.

In 13 of 23 patients with multiple sclerosis, studied with computerized tomography (C.T.) the lateral ventricles were found to be enlarged. In 10 cases sharply demarcated areas of low density were seen in the periventricular region, presumably representing 'plaques'. Cerebral atrophy was more frequent in those patients with a longer duration of the disease, with more exacerbations and with a greater degree of disability. Some areas of low density on C.T. scans correlated with clinical symptoms.


Studies on the mechanism of the positive inotropic action evoked by epinine on the rabbit isolated papillary muscle at different rates of beating.

1. Effects of epinine on cyclic AMP and contractility were investigated in rabbit papillary muscles driven at a rate of 0.5 or 2.0 Hz. 2. When the frequency of stimulation was increased from 0.5 to 2.0 Hz, the log dose-response curve for the positive inotropic effect of epinine was displaced to the left, whereas the maximum of the developed tension was not changed. 3. At both frequencies phentolamine (1 mumol/l) shifted the lower part of the log dose-response curve for epinine to the right, whereas pindolol (30 nmol/l) affected mainly the upper part. In the presence of both alpha- and beta-adrenoceptor antagonists, the whole curve was shifted to the right in a parallel manner. However, cocaine (30 mumol/l) did not significantly influence the log dose-response curve of epinine. 4. At 0.5 Hz a submaximal effective concentration of epinine (100 mumol/l) led to an approximately 100% increase of the cyclic AMP level after 60s; the same increase of the cyclic AMP level was induced at 2.0 Hz by one-third the concentration of epinine (30 mumol/l). 5. Phentolamine (1 mumol/l) did not affect the increase of the cyclic AMP level evoked by epinine, whereas pindolol (30 nmol/l) completely depressed it. 6. The present results indicate that epinine produces its positive inotropic effect through direct stimulation of myocardial alpha-adrenoceptors as well as beta-adrenoceptors, depending upon the concentration: in lower concentrations it acts mainly on alpha-adrenoceptors, whereas in higher concentrations it acts predominantly on beta-adrenoceptors. The positive inotropic effect through beta-adrenoceptor stimulation is mediated by cyclic AMP, while that through alpha-adrenoceptors is not.


Prochlorperazine in childhood: side-effects.

The side-effects of prochlorperazine (CompazineR StemetilR) were traced in thirty-six children up to the age of sixteen years and compared with those collected from literature. These side-effects appeared to be predominantly neurological and were independent of the dosage. Impaired consciousness, dyskinesia, pyramidal signs and hypertonus were the main neurological manifestations. Dyskinesia was the most frequent sign. These side-effects disappear spontaneously after discontinuation of the drug. Instant cure can be acquired by intra-muscular administration of orphenadrine (DisipalR).


Oculopharyngodistal myopathy with early onset and neurogenic features.

Some clinical variants of oculopharyngeal dystrophy are known; a rare form is described in this article: the early-adult form of oculopharyngodistal myopathy. The diagnosis was made in 2 patients ((brother and sister) on the grounds of extensive clinical, biochemical and morphological (microscopical, histochemical and submicroscopical) investigations. Although oculopharyngeal dystrophy is generally considered to be a purely myogenic condition, in one of our patients some neurogenic indications were found (EMG and biopsy). The general picture, however, was that of a myopathy with the characteristic morphological signs of oculopharyngeal dystrophy in the skeletal musclebiopsy.


Glomus jugulare tumours in South Wales--a twenty-year review.

Fourteen patients with glomus jugulare tumours presenting over the last 20 years in South Wales are reviewed. All were treated with radiotherapy and only two died, both of whom had radiological evidence of extensive intrapetrosal spread. All the remainder had good relief of symptoms and are at present free from evidence of progressing tumour.


The absence of 24,25-dihydroxycholecalciferol in anephric patients.

1. In subjects with normal renal function there was a strong positive correlation between serum concentrations of 25-hydroxycholecalciferol and 24,25-dihydroxycholecalciferol, as measured by competitive protein-binding assay. 2. The 24,25-dihydroxycholecalciferol concentration was about 7% of the prevailing 25-hydroxycholecalciferol concentration. 3. In contrast, sera from anephric patients contained very low or undetectable amounts of 24,25-dihydroxycholecalciferol even after the serum 25-hydroxycholecalciferol concentrations in these patients had been elevated by oral administration of 25-hydroxycholecalciferol. 4. In a further group of anephric patients, all having normal serum 25-hydroxycholecalciferol concentrations, no radioactively labelled 24,25-dihydroxycholecalciferol was formed from an injected pulse dose of [3H,14C]cholecalciferol. 5. These results indicate that in man the kidney is the major site of 24-hydroxylation of 25-hydroxycholecalciferol.


Carcinomas and other lesions of the liver in mice ingesting organochlorine pesticides.

Carcinomas of the liver were readily induced in male and female mice given organochlorine pesticides orally. The carcinomas were predominantly hepatocellular, with a few being cholangiocellular, and varied from well differentiated to poorly differentiated to undifferentiated. They are capable of invading, metastasizing, and killing the mice. Hemangiosarcomas, leimyosarcomas, and reticulum cell sarcomas were also occasionally seen in mice receiving these chemicals. Hepatic vein thrombosis and hepatic necrosis was observed in some mice.


Adrenal function testing.

Glucocorticoid stimulation and suppression tests are essential to the definitive diagnosis of diseases of the hypothalamic-pituitary-adrenal axis, because they document abnormal physiologic control of hormonal secretion. Similarly, diseases of the renin-angiotensin-aldosterone axis are diagnosed by mineralocorticoid stimulation and suppression testing. [Ed. Note: See Moore TJ, Williams GH: Adrenal causes of hypertension, in this issue.] Unlike tests of glucocorticoid function, testing of the renin-angiotension-aldosterone system is more complicated, because knowledge of posture and dietary sodium are necessary to interpret the results. However, measurement of the tropic hormone renin and plasma levels of aldosterone can be accurately made, allowing precise definition of this system. Errors are most commonly encountered when dynamic tests of cortisol output are performed in patients taking medications that may interfere with the assays or with the metabolism of the administered compounds, such as dexamethasone or metyrapone. Abnormal, spurious values may also be obtained in some individuals who do not have adrenocortical hyperfunction if they are very obese or if testing is performed in a setting of clinical stress. Careful attention to these pitfalls will avoid errors and allow the clinician to arrive at the correct diagnosis.


Dithiocarbamate therapy for nickel dermatitis.

Increased internal exposure to nickel can cause an exacerbation of nickel contact dermatitis. Nickel ions are chelated by diethyldithiocarbamate (DDC) and thereby inactivated. An oral dose of about 1 g DDC/day was given to a patient. The nickel excretion in the urine increased about tenfold; the nickel elimination in scalp hair did not increase. The slightly negative nickel balance did not exhaust the nickel content of the organs appreciably with a dose of 1.2 g DDC/day for 2 months. At the end of this experiment patch tests with nickel sulphate were still positive though less local therapy was needed, and the cross correlation between the activity of the eczema and the nickel concentration in the urine had lost its former periodicity. It is therefore not yet possible to conclude whether or not DDC may be really of help in the very nickel hypersensitive patient by reducing the exposure to nickel originating in food and other environmental sources.


Enhancement of simian virus 40 uptake by permissive, semi-permissive and non-permissive cells.

The uptake of simian virus 40 (SV40) by cells that are both non-permissive for virus replication and resistant to virus infection could be enhanced markedly by infecting the cells in presence of DEAE-dextran. Virus uptake by semi-permissive and permissive cells was also enhanced by DEAE-dextran. Optimum enhancement of virus uptake occurred at 100 microgram of DEAE-dextran per ml and under the conditions employed, the polycation was not toxic to cells. The increased cellular uptake of virus may result from the uptake of virus aggregates formed in the presence of DEAE-dextran.


Lymphocyte stimulation by herpes simplex virus (HSV) antigens. 1. Culture conditions, cell fractionation and presentation of HSV antigens.

Optimal culture conditions for rabbit lymphocytes were established. Inclusion of 2-mercaptoethanol in the medium greatly enhanced responses. Lymphocytes from the blood of HSV-immunized rabbits responded specifically in vitro to heat-killed (56 degree, 60 min) or UV-inactivated HSV preparations. A UV dose of 13,392 ergs/mm2 was the most suitable inactivating dose. Oral intramucosal injection was the most effective way of generating blood lymphocyte responses to HSV, but animals immunized in tradermally or in tramuscularly with HSV in Freund's adjuvant also produced antigen-reactive cells after five or more immunizations. There were differences in the shape of the dose-response curves to various HSV1 mutants which were probably due to differences in the production of "stimulatory" and inhibitory antigens. Appreciable lymphocyte stimulation could be obtained with tissue culture fluid and enveloped virus antigens. Lymphocytes from HSV1-immune animals were also responsive to HSV2 antigens.


Karyotype evolution of the simian virus 40--transformed human cell line LNSV.

We have used trypsin-Wright's banding ("GTG-banding") to analyze the chromosome content in two sublines of the SV40-transformed human cell line LNSV, derived from fibroblasts of a patient with HPRT deficiency. Both LNSV sublines (GM-847 and "LNSV") were heteroploid and showed considerable numerical and structural chromosome variability. Nineteen rearranged chromosomes which were observed at high frequency have been set aside as "marker chromosomes," and their probable derivation from normal human chromosomes has been described in PARIS CONFERENCE (1971) nomenclature. Heterogeneity within these uncloned sublines appears to increase with time in culture, and no evidence was found for evolution of a karyotypically stable cell population. The results are of general significance for cell genetic studies using established cell lines.


Serial fiberoptic bronchoscopy during chemotherapy for small cell carcinoma of the lung: early detection of patients at high risk of relapse.

Serial fiberoptic bronchoscopic examinations were performed during intensive chemotherapy with a combination of drugs in 77 previously untreated patients with small cell carcinoma of the lung. Before treatment, bronchoscopic examination revealed evidence of cancer in 93 percent (70) of the 75 patients studied at that time, including 8 percent (six) in whom the tumor was not evaluable on the chest x-ray film. After therapy was initiated, 36 percent (29) of the 81 procedures performed in patients with a complete response radiographically and 62 percent (33) of the 53 bronchoscopic procedures in those with a partial response or no response showed evidence of tumor. In both of these groups, patients with abnormal findings on endoscopic examination had a much higher rate of relapsing tumor of the chest within a 12-week period. Progression of intrathoracic tumor was first detected solely by bronchoscopic examination in 22 percent (seven) of the 32 episodes of progression. In our hands, repeated fiberoptic bronchoscopic procedures during chemotherapy for small cell carcinoma have yielded information not apparent from the chest x-ray film in a significant number of patients.


The significance of a cytologically negative pleural effusion in bronchogenic carcinoma.

Bronchogenic carcinoma complicated by ipsilateral cytologically positive effusion is considered unresectable. Bronchogenic carcinoma with cytologically negative effusion, even if bloody, has also been thought by many to be unresectable. Seventy-three patients with bronchogenic carcinoma and ipsilateral cytologically negative effusions were studied. Sixty-six underwent exploratory thoracotomy for staging or therapy; five had pleural biopsies and two had mediastinoscopies, all disclosing metastatic carcinoma. Four of the 73 patients (5.5 percent) had surgically resectable disease. They have remained free of disease for 3, 6, 7, and 14 years. Sixty-nine patients (94.5 percent) had unresectable carcinoma with metastases. Seventeen (94 percent) of 18 patients with bloody effusions had unresectable cancer. Carcinoma was resected in one patient with cytologically negative bloody effusion, and the patient remained free of disease during the 14-year follow-up period. Unresectability must be documented surgically in these patients to exclude those in whom curative resection can be performed.


[The problem of early diagnosis in discrete peripheral nerve lesions. Comparative study of F-wave latency and conventional motor nerve conduction velocity in healthy persons and diabetics (author's transl)].

In 71 healthy subjects and in 38 diabetics the shortest F-wave latencies were evaluated for the n.medianus, n.ulnaris, n.tibialis and n.peronaeus. The motor nerve conduction velocities calculated hereby for the upper and lower extremities were compared with the corresponding distal motor conduction velocities measured in the conventional manner. The comparison of the 2 differently defined motor conduction velocities in healthy and diabetic subjects yielded a better separation between the normal and pathological range in all 4 nerves with the method based on the F-wave latency. For the group of healthy subjects the statistical basis of a preliminary normal range of the F-wave latency was established and nomograms were provided on the basis of the 99% confidence limits which--knowing the length of the arm or leg--open the possibility to derive the corresponding motor conduction velocity.


Effects of thyroliberin, triiodothyronine and cyclic guanosine 3'5'-monophosphate on the volume of nuclei of thyrotropic cells in rat anterior pituitary in vitro.

Influence of TRH, T3 and cGMP on the volume of thyrotropic cells nuclei in rat pituitary in vitro has been examined. A significant increase of the volume of thyrotropic cells nuclei in the culture exposed to TRH, in another one exposed to cGMP and in the third one exposed to both TRH and cGMP together have been observed. The mean volume of cells nuclei within the group exposed to T3 was significantly lower, when compared to the control. T3 inhibited the TRH-induced increase of the volume of thyrotrophs nuclei. The obtained results suggest that the role of cGMP and T3 in the regulation of the thyrotropic cell function are independent and rather opposed. Since TRH increases the level of cAMP in the anterior pituitary, additive effect of TRH and exogenous cGMP support the assumption that both the cyclic nucleotides within the thyrotropic cell act in the same direction.


The neurohypophysial vasopressin content as influenced by modified cholinergic or adrenergic transmission during long-term dehydration in the white rat.

In rats dehydrated up to 12 days the neurohypophysial vasopressin content was determined by Dekański's method. Carbamylcholine inhibited somewhat the vasopressin depletion in the neurohypophysis, but not earlier than under severe dehydration (8th and 12th day). A single dose of atropine given 24 h prior to sacrifice to not dehydrated animals resulted in a diminution of the vasopressin content in the neurohypophysis; in animals dehydrated for four days and parallely atropinized the decrease of the neurohypophyseal vasopressin content was, on the contrary, considerably inhibited. Under severe dehydration, the treatment with atropine did not change the vasopressin stores in the neural lobe. Phenoxybenzamine inhibited the vasopressin depletion in the neural lobe following four days of dehydration. Under severe dehydration, amphetamine potentiated the effect of osmoreceptor stimulation. It is supposed that impulses of osmoreceptor origin are of some importance in determining the vasopressin release following changes of cholinergic or adrenergic transmission.


[ACTH-, glucose- and lactate-content of swine plasma during insulin-induced hypoglycemia].

The present investigation was undertaken to determine the content of ACTH, glucose and lactate in plasma of 4 pigs (body weight 82--118 kg) during a circadian period and during an insulin hypoglycemia test using 1 IU/kg in 3 pigs (body weight 96--118 kg) and 4 pigs (body weight 20--30 kg). The plasma ACTH level at rest was 57 +/- 27 pg/ml (Mean +/- SE) for all samples in all animals during a circadian period. Significant diurnal changes were not observed. During insulin-induced hypoglycaemia plasma ACTH rose from a mean (+/- SE) basal level of 35 +/- 15 to a maximum of 673 +/- 100 pg/ml at 60 min in heavier pigs and in lighter pigs to 395 +/- 153 at 30 min and 403 +/- 145 pg/ml at 120 min. Initial ACTH responses were evident 30 min (heavier pigs) and between 0 and 15 min (lighter pigs) after insulin administration. Plasma glucose decreased from a mean (+/- SE) basal level of 80 +/- 10 to a minimum of 6 +/- 1 mg/100 ml at 60 min (heavier pigs) and from 88 +/- 3 to 16 +/- 4 mg/100 ml at 60 min (lighter pigs). After its minimum level the glucose concentration showed a slower increment in the heavier pigs as compared to lighter animals. Plasma lactate rose from a mean (+/- SE) basal level of 19 +/- 10 to a maximum of 76 +/- 42 mg/100 ml at 120 min (heavier pigs) and from 12 +/- 3 to 37 +/- 16 mg/100 ml at 150 min (lighter group). In accordance with the changes in the blood plasma levels of ACTH, glucose and lactate, the clinical symptoms of hypoglycaemia in heavier pigs were more intensive.


[Daily cyclic changes in thyroid cAMP in chickens (Gallus domesticus)].

48 White Leghorn cockereles (age: 4 weeks) were examined for daily alterations of thyroidal and plasma cAMP-concentrations by means of a radioimmunoassay. In the morning and in the evening the thyroidal cAMP-concentration was elevated. This elevation was accompanied by a decrease of thyroid-weight. Alterations of blood cAMP-concentration were without recognizable effect on thyroid function. It is assumed that mediated by the effect of the second messenger more thyreoglobulin is released at these times.


Benign tumors of oesophagus: value of endoscopy.

Endoscopic examination carried out in 20,000 patients allowed 7 benign tumors of the oesophagus to be detected. Four of them were intramural tumors corresponding to leiomyomas. Guided biopsies are not sufficient to assure an accurate diagnosis, but a mucosal lesion, particularly a carcinoma, should be ruled out. Endoscopic resection of the 3 intraluminal tumours was performed without incidents, allowing an accurate histologic diagnosis and a reasonable therapeutic approach. Endoscopic supervision is necessary in case of epithelial tumors.


Epileptogenic agents applied to trigeminal ganglia: absence of neuronal hyperexcitability.

Chronic application of alumina cream to the trigeminal ganglion in 10 cats failed to produce a long-lasting behavioral syndrome of facial dysesthesia. Histologic and electron microscopic analysis demonstrated morphologic similarities between these ganglia and primate neocortical alumina cream epileptic foci. None of 87 ganglia neurons recorded extracellularly exhibited evidence of intrinsic hyperexcitability, i.e., abnormal spontaneous or physiologically evoked activity or any significant differences in threshold of responses to antidromic or orthodromic electrical stimulation, compared to 67 normal ganglion cells. Furthermore, topical application of penicillin to normal ganglia failed to produce abnormal activity in 42 neurons tested. These data suggest that neuronal somata lacking either dendrites or postsynaptic membranes, or both, do not develop abnormal firing behavior when challenged with these two epileptogenic agents.


Enhancement of alcohol withdrawal seizures with 6-hydroxydopamine.

The relationship between central catecholamine neurons and alcohol withdrawal seizures was studied in rats pretreated with 6-hydroxydopamine to selectively destroy the central catecholamine nerve terminals. The alteration of alcohol withdrawal seizures was manifested by (1) a higher percentage of the rats developing the seizures, (2) an earlier onset of seizures, and (3) a greater severity of seizure activity. Seizure status was exhibited in 25% of the 6-hydroxydopamine-treated rats.


Preferential charging of tRNA-Met-f in Escherichia coli K12.

The charging of tRNA-Met-f and tRNA-Met-m in vivo and in vitro and initiation of polysomes during methionine limitation were studied in two strains of Escherichia coli K12. In the wild-type strain the distribution of polysomes as well as the kinetic parameters of methionyl-tRNA synthetase indicate preferential acylation of tRNA-Met-f. This preferential charging of tRNAM-et-f does not take place in a mutant strain which is also defective in initiation of polysomes during methionine limitation.


Synthesis and breakdown of pppA2'p5'A2'p5'A and transient inhibiton of protein synthesis in extracts from interferon-treated and control cells.

Incubation of the mouse L-cell-free system with a concentration of pppA2'p5'A2'p5'A [(2'-5')An] just sufficient to inhibit protein synthesis results in formation of a high-molecular-weight, heatlabile inhibitor and enhanced ribonuclease activity and in the rapid breakdown of (2'-5')An to ATP. The (2'-5')An-enhanced ribonuclease activity is also unstable and in the absence of a (2'-5')-An-regenerating system inhibiton of protein synthesis is transient. Although interferon treatment enhances the synthesis of (2'-5')An, the rates of degradation of (2'-5')An and levels of activatible nuclease are similar in extracts prepared from control or interferon-treated cells. Interestingly, the sensitivity of different cell-free systems to (2'-5')An, varies with the source of the cell-free systems and with the methods used in their preparation. There is, however, no obvious correlation between the sensitivities of the system and the rate of breakdown of (2'-5')An. The significance of these results is discussed in relation to a possible control function for the (2'-5')An system in both interferon-treated and control cells.


Comparative experimental study of myocardial energy metabolism during ventricular workload induced by chronic stenoses of ascendent aorta and pulmonary artery.

In this paper, an attempt was made to describe the alterations of the myocardial energy metabolism following moderate stenoses of aorta or pulmonary artery. Biochemical investigations regarding the main high energy phosphates and a large series of dehydrogenases in the myocardium of the overloaded ventricles have revealed the following facts: (1) the myocardial CP-CPK system appears to be more labile than the myocardial AMP-ATP system in response to a ventricular overload; (2) the changes in the amounts of myocardial high energy phosphates seem to be more important in the right chronic cardiac overload than in the left one; (3) before the overloaded ventricle becomes insufficient, the myocardial dehydrogenase system elicits no alteration, suggesting that the oxidative phosphorylation is not affected; (4) there is a closer relationship between the coronary flow and pressure and the myocardial energy metabolism than that existing between this metabolism and the tension of the myocardial fibres following a persistent increase in the systemic arterial resistance.


Measurements of total and regional tumor blood flow and organ blood flow using 99Tcm labelled microspheres. An experimental study in rats.

Cardiac output, regional tissue and arterial tumor blood flow were studied in rats, with 99Tcm labelled microspheres using a reference sample simultaneously drawn as the spheres were injected. Both cardiac output and regional blood flow can be estimated in absolute values (ml X min-1 X g-1). Cardiac output and normal tissue blood flow were not affected by tumor growth in the liver or subcutaneously. In small tumors total blood flow was inversely proportional to tumor size, a finding similar in all 3 tumors studied. A relation found in liver as well as in subcutaneously implanted tumors. Blood flow in subcutaneous tumors was greater than muscle or skin blood flow. BP sarcoma had higher blood flow than the hepatoma and the adenocarcinoma. In big tumors the peripheral blood flow was larger than that in the centre of the tumor.


Incidence and natural history of renal stone disease and its relationship to calcium metabolism.

In three different general health surveys, including more than 17,000 individuals, the prevalence of renal stone disease was over 10% of all males and 3% of all females. Stones are thus far more common in the population than is generally appreciated from hospital statistics. Each year approximately 1% of the entire adult male population experiences renal stones. This figure appears to increase rapidly and is now at least twice as high as only 20 years ago. Stone formers as a group had a raised urinary excretion of calcium, but were not particularly accumulated in the upper part of the normal urinary calcium distribution. This pattern was generally caused by an intestinal hyperabsorption of calcium, apparently not dependent on the action of parathyroid hormone. The prevalence of stone disease was closely related to the urinary calcium output.


alpha-Adrenergic modulation of hypothalamic self-stimulation: effects of phenoxybenzamine, yohimbine, dexamphetamine and their interactions with clonidine.

The alpha-adrenoceptor agonist clonidine (12.5--50.0 microgram/kg) produced a dose-dependent increase in the latency to initiate lateral hypothalamic stimulation. The insurmountable postsynaptic alpha-adrenoceptor antagonist phenoxybenzamine (0.2-0.8 mg/kg) had no effect on self-stimulation by itself, but potentiated the inhibitory effects of clonidine. The fact that the concurrent escape behavior to the intracranial stimulation was unchanged by either clonidine or the phenoxybenzamine-clonidine combination suggests that the inhibition is specific to the rewarding component of hypothalamic stimulation. Yohimbine (0.5--2.0 mg/kg) produced a dose-dependent increase in both response latencies. This lack of behavioral specificity may reflect yohimbine's wide range of pharmacological activity, Dexamphetamine (0.25--0.50 mg/kg) reversed clonidine's inhibition of self-stimulation reward in a specific and dose-dependent fashion. This reversal could be blocked by previous inhibition of catecholamine synthesis with alpha-methyl-p-tyrosine. These data support the concept that the alpha-adrenoceptors play a critical role in the modulation of hypothalamic self-stimulation reward. They further suggest that the inhibitory effects of clonidine on self-stimulation reward represent an agonist effect on presynaptic alpha-adrenoceptors.


Variations in endogenous phosphodiesterase activator content of rat aorta.

The variations with age and blood pressure in the cyclic nucleotide phosphodiesterase activator content of rat aorta were studied. Activator content increased with age and was less in mineralocorticoid-hypertensive than in control rats. These variations were more pronounced in the particulate than in the soluble fraction of aorta. The data do not demonstrate any relationship between the activator level and blood pressure, but they suggest that the activator may modulate cyclic nucleotide metabolism in rat aorta.


Effect of extracellular Na on isoproterenol-induced cAMP levels in depolarized uterus.

In the present study, the absence of a quantitative correlation between isoproterenol induced relaxation of uterine strips and tissue cAMP levels was demonstrated by using three depolarizing media: 127 mM KCl (0 NaCl), 47.5 mM KCl (NaCl), and 47.5 mM KCl (80 mM NaCl). While the degree of relaxation by isoproterenol was similar in all three media, isoproterenol (10(-4)M) increased cAMP by 100% in Na+ free depolarizing media, by 337% in Na+ containing depolarizing medium and by 600% in normal non-depolarizing medium. After pretreatment of the tissue with 10(-5) M D-600, 10(-4) M isoproterenol increased cAMP levels by 600% in all three depolarzing media. Studies using 10(-8) M isoproterenol produced qualitatively similar results. cGMP levels did not change significantly in any of the above studies. Na+ appears to be producing its effect on isoproterenol induced increase in cAMP levels indirectly by reducing the increase in intracellular Ca2+ concentration known to occur with depolarization.


Effect of dibutyryl cyclic AMP and analogs on the rate of contractions of myocytes in culture.

2O, 6N-butyryl, 3', 5'-cyclic monophosphate (dibu cAMP) when added to fetal rat heart cells in culture inhibits myocyte contraction. This inhibition is 100, 84 and 51% complete when the dibu cAMP concentration used is 2, 0.2 and 0.02 mM, respectively. The potency of dibu cAMP derivatives in myocyte contraction inhibition follows the order, dibu cAMP greater than 6N-bu cAMP greater than 2O-bu cAMP = AMP greater than butyrate. The inhibition caused by the first three chemicals is greater than 70%.


Acetylcholine synthesis and its dependence on nervous activity.

The findings discussed in this paper mainly derived from studies on salivary glands, serving as model organs, indicate that the capacity to form the neurotransmitter acetylcholine, as judged by the activity of choline acetyltransferase, is influenced by the traffic of nerve impulses, as a long term effect. In the glands, choline acetyltransferase seems to be exclusively localized to the cholinergic nerves. In the postganglionic parasympathetic nerves of the glands, the activity of choline acetyltransferase decreases when the flow of secretory impulses in these nerves is abolished or reduced either by isolating the nerves from the central nervous system, surgically or pharmacologically, or by diminishing the reflex activation of the glands from the mouth. The opposite occurs when the reflex activation of the salivary glands is enhanced, i.e. the activity of choline acetyltransferase increases. Observations on various other organs are quoted in support of the view that the traffic of nerve impulses is of importance for the activity of the enzyme. An increase in choline acetyltransferase activity also occurs in some salivary glands after sympathetic denervation. This puzzling observation is discussed in relation to impulse traffic. Increased nerve impulse traffic and collateral sprouting seem to be responsible for the rapid restitution of choline acetyltransferase activity from a low level in an organ partially deprived of its cholinergic nerve supply.


Correlation between 3',5', c-AMP levels and thyrotropin in separated rat pituitary thyrotropic cells.

The correlation between 3',5', c-AMP levels, TSH content and secretion of separated thyrotropic cells was studied. Incubation of the separated cells with 1, 10 and 100 ng of TRH does not change the 3',5',c-AMP levels, despite the significant rises of the TSH level. Dibutyryl c-AMP causes rise in TSH content, with no indication of its secretion. PGE2 10(-5) increased 3'5',c-AMP levels with no change in the content or secretion of TSH in separated thyrotropic cells.


Cyclic AMP is a likely mediator of ovulation in the tsetse fly.

Ovulation in tsetse flies is normally induced by mating, but virgins can be stimulated to ovulate with an injection of dibutyryl cyclic AMP, cholera toxin (a cyclic AMP generator), or aminophylline (a phosphodiesterase inhibitor). Thus, elevation of cyclic AMP is a likely link in the events leading to ovulation.


Naloxone and intestinal motility.

It was supposed that the inhibition of intestinal peristalsis seen in animals and humans after abdominal surgery might be related to the release of endorphins, endogenous opiate receptor agonists, caused by the surgical stress and pain. However, naloxone, a potent morphine and endorphin antagonist, failed to block this peristaltic inhibition in rats, which leaves the mechanism of this inhibition, and thus the function of intestinal endorphins, still very much in doubt.


Cadmium effects on bone and dental tissues of rats in acute and subacute poisoning.

Injection of cadmium chloride to rats fed by normal calcium diet induced distrubances of tibia and incisor hard tissues metabolism. It is suggested that there may be the possibility of direct cadmium actions on those hard tissues metabolisms.


Can the binding of GABA, glycine and beta-alanine to synaptic receptors be determined in the presence of a physiological concentration of Na+?

Bicuculline- and strychnine-sensitive components of the binding of GABA, glycine and beta-alanine, which can be demonstrated in the presence of a physiological concentration of Na+, might be related to synaptic receptors.


Influence of dithiocarb on the biliary excretion of paracetamol and bilirubin in rats.

In bile-fistula rats, the biliary elimination of conjugated paracetamol and conjugated bilirubin is diminished by simultaneous administration of dithiocarb. This dithiocarb effect could be the result of the interference with the glucuronidation of the compounds.


Relationships between neural crest cells and catecholamine in suckling mice.

In suckling mice injected i.p. with a 27 mg/kg dose of L-hydrochloric acid isoproterenol, multiple neural crest tumors developed and cell death of neural crest cells occurred. It is speculated that neural crest derivatives may be beta-receptor cells and contain regulatory units of neurotransmission mediated through cyclic AMP.


[Azotized emetine analogues: synthesis and evaluation of the anti-amebic and anti-tumoral action of aza-3 emetine and attempted synthesis of aza-2 emetine].

The synthesis of the N,N-bis-(acetylhomoveratrylamido)-1-hydroxymethyl and 1-carboxypropylamines (III a) and (III b) is described. Until now, these molecules could not been cyclized to the corresponding isoquinoline compounds. An attempted synthesis of 3-azaemetine by condensing homoveratrylamine with acetonedicarboxylic acid or its esters did not yield the expected diamide (XIV). However, by subjecting 1,3-bis-(1,2,3,4-tetrahydro-6,7-dimethoxy-1-isoquinolyl)acetone (XV) to a reducing aminoalkylation, one obtained the corresponding ethylamino derivative (XVI) which could be cyclized through Mannich reaction to give 3-azaemetine. The pharmacological screening showed 3-azaemetine to be less toxic than emetine in the mouse and without antiamebic and antitumor activity against P 388 Leukaemia in the mouse (25).


Distribution of Na+-pump sites in transporting epithelia.

There are three techniques for the localization of intraepithelial Na+, K+-ATPase (usually equated with the Na+-pump) that offer reasonable specificity and resolution: the nitrophenylphosphatase assay of Ernst, the immunoferritin procedure of Kyte, and the radioautographic localization of tritiated ouabain as developed by Stirling. These have now been applied to a wide range of epithelia covering the four classes of interest here: isotonic and hypertonic absorbers and isotonic and hypertonic secretors. A review of published results reveals that in every case (except for the choroid plexus) the enzyme is preferentially located on the basolateral surface of the transporting epithelial cells so that a simple correlation  of structure and function in terms of the Koefoed-Johnsen and Ussing hypothesis does not seem possible. With little dispute that this enzyme is, nonetheless, the probable site for conversion of metabolic energy to transport-related work, we summarized as well the more macroscopic structural characteristics of epithelia which serve to typify each of the four classes in terms of the direction and tonicity of transported fluid. The apparently systematic differences in cell shape and cell-cell junctions that are summarized here may well be an important consideration for the development of a useful holistic theory with which to explain the transepithelial transport of salt and water.


Hormonal regulation of cAMP-independent protein phosphokinase activities: thyroxine and cortisol control of enzymes from rat liver cytosol.

Thyroxine control of cAMP-independent histone and casein phosphokinase activities was studied in thyroidectomized rats treated with thyroxine. All activities were evaluated in the presence of a thermostable inhibitor of cAMP-dependent enzymes. Cytosol enzymes can be resolved by sucrose gradient ultracentrifugation into three peaks of histone kinase activity (3.2S, 5S and 7.2S) and two peaks of casein kinases (3.6S and 7.1S). Neither thyroidectomy nor subsequent treatment of operated animals with thyroxine modifies the total histone kinase activity estimated, either in total cytosol or after its fractionation by the sucrose gradient ultracentrifugation. The activity ratios of different peaks were, however, changed. Casein kinase activity was significantly decreased after thyroidectomy (about 50%). Subsequent treatment with thyroxine restored this activity to its initial value. Sucrose gradient ultracentrifugation analysis showed that thyroxine action on the casein kinase activity is very specific. Only molecules that sediment in the 9S region were significantly stimulated by the hormone. Cortisol action on the casein kinase activity was studied in adrenalectomized animals treated with hormone for 24 h. Cortisol decreases the total casein kinase activity by about 30%. Sucrose gradient ultracentrifugation analysis showed that the population of molecules sedimenting at about 9S was the most sensitive to cortisol. The above data show that both thyroxine and cortisol control, in a selective way, the activities of cAMP-independent protein kinases. The same kinase molecules can be under double control by two different hormones that have opposite effects.


Prolactin receptor on dissociated mammary epithelial cells at different stages of development.

Properties of prolactin receptors were measured by monitoring [125I]prolactin binding to specific receptor sites on collagenase-dissociated mammary epithelial cells of virgin, pregnant and lactating mice. On a Scatchard plot the data generated a straight line and the estimated dissociation constant (Kd) and number of receptor sites on lactating cells were 0.9 x 10(-9) and 1540 per cell. The [125I]prolactin binding was inhibited in presence of unlabeled prolactin and other lactogenic polypeptide hormones, but not by nonlactogenic polypeptide hormones. The [125I]prolactin binding was sensitive to pronase and trypsin but not to DNAase, RNAase and hyaluronidase. Scatchard plot analysis further showed that while the number of receptors on mammary cells was variable at different stages of endocrine regulated developmental changes of the gland, Kd of the hormone--receptor complex generally remained similar. The high level of prolactin receptors on mammary cells of virgins was reduced during pregnancy and the lactating mammary cells showed a highly elevated level of prolactin receptors. The results demonstrate that specific prolactin receptors can be measured on collagenase dissociated mammary epithelial cells and this method permits a direct assessment of the number of receptors on a per cell basis rather than indirect estimates, based on average DNA or protein content of the tissue, composed of heterogeneous cell types.


Pregnenolone biosynthesis in isolated cells of Snell rat adrenocortical carcinoma 494.

Adrenocorticotrophin (ACTH) produced an insignificant stimulation of pregnenolone biosynthesis from endogenous precursors in isolated cells prepared from the rat Snell adrenal carcinoma 494. On the addition of 25-hydroxycholesterol, the rate of pregnenolone synthesis increased 10-fold. These results, noting also the very low cholesterol content of the tumor cells, suggested that lack of cholesterol was responsible for the poor steroidogenic response of the cells to ACTH. Endogenous pregnenolone production was sensitive to cytochalasin B as well as cycloheximide. However, pregnenolone synthesis after the addition of 25-hydroxycholesterol was not affected by these inhibitors. Removal of cycloheximide from the cells resulted in the immediate restoration of the initial rate of pregnenolone synthesis from endogenous precursors. This suggested that cycloheximide was interfering with the action of a stable activated intracellular messenger.


Virus-induced diabetes in mice: a quantitative evaluation of islet cell population by immunofluorescence technique.

The endocrine cell populations of pancreatic islets in encephalomyocarditis (EMC)-virus infected mice were assessed quanititatively by immunofluorescence using specific antisera against 4 islet hormones. A marked reduction of the volume of insulin-containing (B-) cells (up to one tenth of control values) was observed at all stages studied in the hyperglycaemic mice. This was accompanied by the inversion of the normal ratio between B- and non B-cells. The volume of the latter cell types was also modified at different time points after infection: glucagon-cells were augmented 14 days after infection; PP-cells were decreased 2--3 days and 21 days after infection; somatostatin-cells decreased to one-fourth of control values in hyperglycaemic animals 21 days after infection. The latter results suggest that non B-cells are also involved in islet reaction to virus infection.


Acid glycohydrolase in Chinese hamster with spontaneous diabetes. IV. Diabetes- and line-dependent variation in plasma enzyme activity.

The activity of five glycohydrolases was measured in the plasma of Chinese hamsters from eight highly inbred lines in the Upjohn colony. The diabetic animals showed elevated activities of plasma beta-D-galactosidase, N-acetyl-beta-D-glucosaminidase and N-acetyl-beta-D-galactosaminidase but similar activities of plasma alpha-D-glucosidase and alpha-D-galactosidase to the nondiabetic animals. Line-specific variation was observed in all five enzymes and anomalies were especially evident in N-acetyl-beta-D-glucosaminidase and N-acetyl-beta-D-galactosaminidase activity. In two diabetic lines, AC and Z, activities of these two enzymes were not elevated although significant correlation was found with blood sugar levels. The pronounced difference in the plasma activity of N-acetyl-beta-D-glucosaminidase in two diabetic lines, XA and AC, did not involve plasma inhibitors or activators, as evidenced by the coincidence in observed and calculated activities in mixed plasma samples, or specific isozymes, concluded from the similar elution profiles on ion-exchange column and thermostability curves. These data suggest that diabetes-related changes in plasma glycohydrolase activities are dictated by genetic factors and may be involved in the development of complications.


The metabolism of cyclic AMP and glucose in isolated islets from Acomys cahirinus.

Glucose-induced cyclic (3H) AMP accumulation, insulin secretory responses and the metabolism of glucose were studied in pancreatic islets from Acomys cahirinus. 27.7 mmol/l of glucose stimulated neither islet cyclic (3H) AMP accumulation nor insulin release during the first 5 min of incubation. Stimulation by glucose of cyclic (3H) AMP was observed after 15 min of incubation and insulin release was markedly stimulated between 15 and 30 min. The utilization of glucose, measured as the production of (3H)2O from (5--3H) glucose was stimulated by glucose after 10 min and proceeded at an apparently linear rate during a 20 min incubation period. In incubations of 5 min, glibenclamide, glucagon or chloromercuribenzene-p-sulphonic acid failed to stimulate islet cyclic (3H) AMP accumulation. 3-isobutyl-l-methylxanthine in a concentration of 1.0 mmol/l was the only agent tested that elevated rapidly (1 min) islet cyclic (3H) AMP. None of the agents tested elicited an insulin secretory response in 5 min incubations. It is concluded that 1) no gross defect is apparent in the utilization of glucose by Acomys islets, 2) the secretory derangement of the Acomys is associated with a delayed cyclic AMP response to glucose, 3) however a decreased level of cyclic AMP cannot be the sole explanation for the delayed insulin secretion in the Acomys.


[Carcinoma of the apocrine glands of the vulva with intraepidermal spreading under the picture of invasive Paget's disease (author's transl)].

The histological findings of Paget's disease of the nipple and of extramammary localisation are well known. The relationship to the so called "adnexal carcinoma" has not become clear to this moment. We report a case of combination of invasive vulva Paget's disease and adnexal carcinoma of the apocrine vulvar glands. Our findings are discussed with the statements of literature about aetiology and pathogenes.


Multiple fibrous xanthomas of tendon sheath.

Fibrous xanthomas of the digital tendon sheath system are a common benign tumour. The varying recurrence rates cited in the literature have usually been attributed to inadequate surgical excision. However, Boyes (1964) has suggested "seeding" as a possible mechanism to explain persistence of the tumour. Our case of multiple fibrous xanthomas at different levels in a single digital sheath system is presented as an example of "seeding" along the tendon sheath.


[Obesity and adipose tissue. 2. Hormonal regulation of adipose tissue metabolism].

The effects of hormones on human adipose tissue are reviewed with respect to the pathogenesis, prevention and therapy of obesity. Insulin. The insulin-resistance in the obese is associated with a decrease of the number of insulin receptor sites, which is likely to be secondary to increased insulin levels. Catecholamines. Human adipose tissue contains alpha- and beta-adrenergic receptors. Alterations in the relation of alpha- and beta-adrenergic responsiveness may be important in the pathogenesis of regional forms of obesity. Gastrointestinal hormones. As opposed to adipose tissue of other species lipolytic effects of gastrointestinal hormones were as yet not clearly demonstrated in human fat cells. Prostaglandins were implicated in the pathogenesis of metabolic obesity. However, the effects of these C-20 fatty acids on human adipose tissue remain to be elucidated. Parathyroid hormone has been shown to possess lipolytic activity in vitro. This property may be important under physiological conditions too. Triglyceride storage diseases and lipomatoses are discussed as models for studying impaired hormonal responsiveness in human adipose tissue.


Photolabile and paramagnetic derivatives of the nucleoside X and of Escherichia coli tRNAPhe.

The synthesis of N3-[3-L-(5-azido-2-nitrobenzamido)-3-carboxypropyl]uridine (4b) and N3-[3-carboxy-3-L-(2,2,5,5-tetramethyl-3-pyrroline-3-carbonylamino)propyl]uridine Npyr-oxyl (4c) starting from the nucleoside X (4a) and the appropriate N-hydroxysuccinimide ester 1 or 2 is described. After acylation of tRNAPhe from E. coli (5a) with 1 or 2, the photolabile tRNAPhe derivative 5b and the paramagnetic tRNAPhe derivative 5c could be isolated. The position of modification in the polynucleotide chain was elucidated by comparison of the ribonuclease II/alkaline phosphatase digestion products of the substituted and unsubstituted tRNAPhe samples, and was identified as being exclusively the amino group of the nucleoside X in position 47 of E. coli tRNAPhe.


beta-Nicotinamide-alpha-adenine dinucleotide. Synthesis and properties of a coenzyme analog.

It was not possible to synthesize beta-nicotinamide-alpha-adenine dinucleotide (NalphaAD) via the NAD pyrophosphorylase-catalyzed reaction of beta-nicotinamide mononucleotide and alpha-adenosine-5'-O-triphosphate. It was therefore prepared by reacting a mixture of beta-nicotinamide mononucleotide and alpha-adenosine-5'-O-monophosphate in aqueous pyridine, using N,N'-dicyclohexylcarbodiimide as condensing agent. NalphaAD can replace NAD in the lactate-dehydrogenase-catalyzed oxidation of lactate to pyruvate with LDH 4M from hog muscle as well as LDH 4H from pig heart. Km values are found to be about one order of magnitude higher for NalphaAD than for NAD. The turnover number for NalphaAD with LDH 4H is similar to that of NAD (95%), whereas it is reduced to one third (35%) in the reaction with LDH 4M. Experiments findings are discussed on the lines of a NAD-equivalent conformation of NalphaAD in the holoenzyme complex.


Lipid-protein interaction in reconstituted cytochrome c oxidase/phospholipid membranes.

Deuterium and 31P nuclear magnetic resonance have been employed in an investigation of the effect of cytochrome c oxidase (EC 1.9.3.1) on the structure of lecithin bilayers. Cytochrome c oxidase was isolated from beef heart mitochondria in lipid-free form and reconstituted as a functional enzyme in bilayers composed of synthetic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine. Two separate reconstitution experiments were performed in which the lipid was selectively deuterated either at the C-5' or at the C-14' segment of the palmitic acyl chain. The phospholipid-to-protein ratio of both reconstituted complexes was 0.74 (mg/mg), corresponding to about 200 molecules lipid per molecule cytochrome c oxidase. The deuterium quadrupole splitting deltanuQ, and the phosphorus chemical shielding anisotropy, deltasigma, of the cytochrome c oxidase-phospholipid recombinants were measured as a function of temperature and compared to the results obtained for the pure lipid membrane without protein for the pure lipid membrane without protein. deltanuQ and deltasigma are highly sensitive to the structural organization of the lipid membrane and these measurements demonstrate that the incorporation of cytochrome c oxidase into phosphatidylcholine bilayers leads to a more disordered conformational state of the lipids. This result can be explained by a rapid exchange between lipids in direct contact with hydrophobic protein and those further away from it (exchange rate greater than 10(4) Hz). The irregular protein surface is sensed by all lipid molecules and induces a more disordered bilayer structure. In contrast to previous interpretations, our measurements do not suggest a special type of boundary lipid.


On the modification of adenosine kinase by thiols.

The catalytic activity of adenosine kinase (EC 2.7.1.20) from yeast is very labile. Even incubation with thiols provokes a loss of two thirds of its enzymatic activity. Concomitantly, two SH-groups appear on the enzyme in addition to the single SH-group already present in the untreated enzyme, the latter being absolutely essential for activity. Treatment of adenosine kinase with thiols does not substantially affect the binding of the substrates adenosine and ATP-Mg2. The reactivity of the two newly formed SH-groups is diminished in the presence of ATP-Mg2, whereas adenosine has no influence. The opposite holds for the reactivity of the single SH-group essential for enzymatic activity. Complete reactivation of the enzymatic activity after incubation of adenosine kinase with thiols can be achieved by reoxidation of the enzyme in presence of high concentrations of adenosine. These observations suggest the notion that adenosine kinase contains an essential SH-group close to the adenosine-binding site and a disulfide bridge near to the binding site of ATP-Mg2, the latter being easily accesible to the reduction by thiols.